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Objective The ongoing COVID-19 pandemic warrants accelerated efforts to test vaccine candidates. To explore the influencing factors on vaccine-induced effects, antibody responses to an inactivated SARS-CoV-2 vaccine in healthy individuals who were not previously infected by COVID-19 were assessed. Methods All subjects aged 18–60 years who did not have SARS-CoV-2 infection at the time of screening from June 19, 2021, to July 02, 2021, were approached for inclusion. All participants received two doses of inactivated SARS-CoV-2 vaccine. Serum IgM and IgG antibodies were detected using a commercial kit after the second dose of vaccination. A positive result was defined as 10 AU/mL or more and a negative result as less than 10 AU/mL. This retrospective study included 97 infection-naïve individuals (mean age 35.6 years; 37.1% male, 62.9% female). Results The seropositive rates of IgM and IgG antibody responses elicited after the second dose of inactivated SARS-CoV-2 vaccine were 3.1% and 74.2%, respectively. IgG antibody levels were significantly higher than IgM levels (P 42 days compared to that at 0–20 days (P18 years), which can be influenced by age and detection time after the second dose of vaccination.

Abnormal mRNAs of the hybrid BCR-ABL gene in the majority of cases initiate the synthesis of proteins with a mass of 210 kDa (p210), 190 kDa (p190), and 230 kDa (p230). Expression of the p210 variant is most common in CML (95% of cases), while the p190 and p230 variants are less common (1-4%). On the contrary, p190 predominates in ALL. Measurement of BCR/ABL gene expression is included in clinical guidelines for the diagnosis of CML and ALL as sequential tests in accordance with their occurrence. At the same time, in the context of primary patients testing with suspected hematological malignancies with a low prevalence of BCR-ABL positive patients in the cohort of examined individuals, sequential testing is associated with low cost-effectiveness. Purpose: approbation of a parallel algorithm for detecting all three (p210, p190 and p230) using the multiplex RT-PCR format implemented in the «BCR/ABL-MULTITEST» reagent kit. We used anonymized blood samples from patients with suspected CML, as well as samples from ALL patients before starting therapy. Testing of blood samples was carried out using two variants of the algorithm: sequential determination of individual BCR-ABL transcripts and parallel determination using the developed set of reagents «BCR/ABL-MULTITEST». To detect the p210 transcript, a commercial kit «AmpliSens® Leukemia Quantum M-bcr-FRT» (Central Research Institute of Epidemiology of Rospotrebnadzor, Russia) was used. Simultaneously, a test was used to detect all three variants of BCR-ABL transcripts using the «BCR/ABL - MULTITEST» reagent kit based on a monochrome multiplex reaction «in one test tube». Reverse transcription were carried out using the REVERTA-L reagent kit (Central Research Institute of Epidemiology of Rospotrebnadzor, Russia) in accordance with the manufacturer’s instructions. Using the reagent kits «BCR/ABL-MULTITEST» and «AmpliSens® Leukemia Quantum M-bcr-FRT» there is a high level of correlation of quantitative results of determining the chimeric transcript BCR-ABL р210 (r = 0.99). When using the proposed parallel algorithm with the primary use of the «BCR/ABL-MULTITEST» reagent kit, out of 95 patients with suspected CML, 9 samples with p210 transcript were identified, one with p190 BCR / ABL, and in one case a transcript variant characteristic of chronic neutrophilic leukemia - p230 BCR / ABL. The estimated cost for detecting one positive case of BCR-ABL when using the parallel diagnostic algorithm «BCR/ABL-MULTITEST» with a focused flow of studies is reduced by about 2 times due to a decrease in the amount of laboratory plastic used and the volume of the reaction mixture, as well as the absence of the need for repeated separate tests to detect p190 and p230. The use of the multiplex PCR-RT test system «BCR/ABL-MULTITEST» allows detecting in one test tube all three main variants of BCR-ABL transcripts - p210, p190, p230 and achieving significant resource savings when examining a cohort of patients with suspected CML and ALL and low frequency of positive samples.

Background: Meningoencephalitis refers to the inflammation of the meninges and brain, and is considered an infectious neurological emergency. Tuberculous meningoencephalitis is caused by Mycobacterium tuberculosis and is the most common form of central nervous system (CNS) tuberculosis. Materials and Methods: CSF samples were collected asceptically and processed with an aim to identify and isolate Mycobacterium tuberculosis from clinically suspected cases of meningoencephalitis and observe the antitubercular drug resistance pattern. Results: The study group included 117 patients clinically diagnosed cases of chronic meningoencephalitis which were subjected to Z-N staining, culture on LJ media. From 117 cases, 21 cases were AFB positive, 20 cases were culture positive for Mycobacterium tuberculosis. Antitubercular drug susceptibility pattern was obtained using commercial kit provided by Himedia. (SL023) Conclusion: Tuberculous meningoencephalitis associated with a high frequency of neurologic sequelae and mortality if not treated promptly. Keywords: Meningoencephalitis, Mycobacterium tuberculosis, Anti tubercular drug susceptibility.

OBJECTIVETrigeminal neuralgia is a debilitating disease that can be treated effectively by a number of modalities. Percutaneous balloon compression rhizotomy of the gasserian ganglion is an important technique that can be offered as a primary or secondary strategy after failure of medical therapy. However, the commercial kit for this procedure was discontinued in the United States in early 2016 and therefore is not currently available. The authors describe a low-cost, effective solution for continuing to offer this procedure using equipment already available in most hospitals.METHODSThe authors provide a detailed equipment list with step-by-step instructions on how to prepare all the necessary items and perform a percutaneous balloon compression rhizotomy.RESULTSThe custom “homemade” kit and technique described have been utilized successfully since June 2016 in 34 patients. The kit is a low-cost alternative, and its application does not add any operative time beyond that required for the previously commercially available kit.CONCLUSIONSPercutaneous balloon compression rhizotomy of the gasserian ganglion is an important technique that should be readily available to patients who are not medically fit for microvascular decompression and need immediate relief of their pain. The alternative kit described here can be assembled easily using equipment that is readily available in most hospitals.

Background: The study of hematic concentrations of alpha1 antitrypsin (AAT) is currently one step in the diagnosis of AAT deficiency. To try to clarify the relevance of the laboratory techniques, we carried out a systematic review of the literature. Methods: Studies evaluating the quantification of AAT in peripheral blood were searched in PubMed in July 2021. The selection criteria included (1) any type of study design that included a quantification of AAT in peripheral blood; (2) studies written in English or Spanish; (3) studies evaluating human beings; and (4) studies involving adults. Results: Out of 207 studies, the most frequently used techniques were nephelometry (43.9%), followed by ELISA (19.8%) and turbidimetry (13.5%). Altogether, 182 (87.9%) cases expressed their results in units of gram, while 16 (7.7%) articles expressed them in units of mole. Only 2.9% articles referred to the standard used, 43.5% articles indicated the commercial kit used, and 36.2% indicated the analyzer used. Conclusions: The technical aspects of these determinations are not always reported in the literature. Journals should be attentive to these technical requirements and ensure that they are included in the works in which AAT is determined in order to ensure a correct interpretation of the study findings.

International audience; Toxoplasmosis can be a life-threatening infection, particularly during pregnancy and in immunocompromised patients. The biological diagnosis of toxoplasmosis is challenging and has been revolutionized by molecular detection methods. This article summarizes the data of a multicenter study involving four centers to assess the performances of a commercial PCR assay as compared with four in-house PCR assays using Toxoplasma gondii standards, 20 external quality control specimens, and 133 clinical samples. This clinical cohort includes well-characterized clinical samples corresponding to different clinical situations: confirmed congenital toxoplasmosis (44 samples), toxoplasmosis in immunocompromised patients (25 samples), and chorioretinitis (5 samples). Furthermore, 59 samples from patients without toxoplasmosis were included as negative controls. The analytical sensitivities of the five methods tested were very similar; and the limit of Toxoplasma DNA detection was around 0.01 T. gondii genome per reaction for all the methods. The overall concordance between the commercial PCR and the four in-house PCR assays was 97.7% (130/133). The clinical sensitivity and specificity were >98% and could be increased for the commercial kit when PCR was performed in multiplicate to detect low parasitic loads. In conclusion, the commercial PCR assay shows suitable performances to diagnose the different clinical forms of toxoplasmosis.

Colic horses show systemic inflammatory response syndrome (SIRS) clinical signs. Procalcitonin (PCT) showed increased circulating levels in sick horses. This study compares plasma PCT concentrations in healthy vs. SIRS negative/positive colic horses over time, and evaluates PCT and SIRS score potential correlation, to verify the usefulness of PCT for the evaluation of SIRS severity. Ninety-one horses were included, 43/91 were healthy, on basis of physical examination, blood work and SIRS score (score = 0), while 48/91 were sick colic horses, classified as SIRS-negative (score &lt, 2) and positive (score ≥ 2). Moreover, a 0–6 point-scale SIRS score was calculated (assessing mucous membrane color and blood lactate concentration). PCT was evaluated at admission, and at 24, 48, 72 and 96 h, using a commercial kit for equine species. We verified by the ANOVA test PCT differences between healthy vs. colic horses, healthy vs. SIRS-negative or SIRS-positive colic horses, at all sampling times, and the correlation between the SIRS score at admission with the SIRS score. Statistically significant differences were detected between healthy vs. all colic horses and between healthy vs. SIRS-positive or negative horses at all sampling times. No correlation was observed between the SIRS score at admission and PCT values. PCT was statistically higher in colic horses compared to the healthy ones, suggesting a role as a biomarker for colic.

Background Avian leukosis (AL), which is caused by avian leukosis virus (ALV), has led to substantial economic losses in the poultry industry. The kit used to detect all ALV-positive chickens in breeder flocks is very important for efficiently controlling AL. However, a new emerging ALV subtype is currently a severe challenge in the poultry industry. Results In this paper, we compared different enzyme-linked immunosorbent assay (ELISA) kits for detecting p27 of ALV in the same batch of meconium samples. Different positive samples were further analyzed by PCR or virus isolation. The results showed that 36 positive samples among the 1812 chicken meconium samples could be detected by a sandwich ELISA (sELISA) kit, but only 17 positive samples could be identified by a commercial kit. To verify this result, cloacal swabs and viruses isolated from the positive chickens (2 days old) were used to detect the presence of p27. The results showed that the positive rate of p27 was 100% for the swabs and 40% for virus isolation. Surprisingly, PCR and sequence analysis revealed that the env gene of ALV in these positive samples belonged to the novel subgroup K (ALV-K). Conclusion These data not only demonstrate the relatively high sensitivity of the sELISA kit but also highlight the challenge of controlling ALV-K.

Background: MicroRNAs (miRNAs) are new generation biomarkers used in oncology, cardiology, metabolic syndrome, obesity or in neurology. miRNAs are short non-coding RNA molecules that regulate gene expression in eukaryotes. Aim: To compare a new commercial method for establishing miRNA (imunoassay) with a commercial kit RT qPCR. Methods: RNA was isolated from whole blood samples obtained from four healthy volunteers. The isolates were liquated and miRNA-93-5p and miRNA-23a-3p were measured independently with commercial hsa-miR-93-5p miREIA and hsa-miR-23a-3p miREIA, and commercial RT-qPCR kits. Results: Both miRNAs had good analytical characteristics, very good correlation with RT qPCR. The results between immunoassay and RT qPCR did not statistically differ. A method based on ELISA was faster (2 h with ELISA vs. 3 h with qPCR) and had lower CV then a method based on RT qPCR (see more text). Conclusion: MicroRNAs from blood or derived fractions are particularly interesting candidates for routine laboratory applications. The immunoassay can be performed on any device that processes the ELISA plates and is therefore available in almost every laboratory.

Serum Osteocalcin (OC) is a biomarker for evaluating bone turnover in humans. Commercial kits of OC in India are imported, hence the associated high cost prohibits their use in routine screening of osteoporosis. The present study describes the development, validation of human OC ELISA and establishes cut off values for its use in screening and management of women at risk for osteoporosis.A sandwich OC ELISA was developed using immuno-reagents prepared indigenously and validated for analytical sensitivity, specificity, accuracy and compared with commercial kit using Bland-Altman method. The utility of OC assay was evaluated by ROC analysis.The new ELISA was sufficiently precise, accurate, matrix-free, sensitive and cost effective. The levels of OC were significantly different in women with osteopenia and osteoporosis (ANOVA, p .0001) compared to women with normal BMD. ROC analysis demonstrated the cut off values of OC11.9 ng/mL for osteopenia and 14.9 ng/mL for osteoporosis. The OC levels had maximum AUC of 0.831 in osteopenia and 0.932 in osteoporosis. Further, OC levels showed significant changes within 3 months in women monitored on therapy.The developed OC ELISA has great potential to be used as a biomarker for routine screening and management of osteoporosis in Indian women.

Fish and seafood products has been commonly targeted for fraudulent activities. For that reason, authentication of fish and seafood products is important to protect consumers from fraudulent and adulteration practices, as well as to implement traceability regulation. From the viewpoint of food safety, authenticity is beneficial to protect public from serious food poisoning incidents, such as due to ingestion of toxic species. Since DNA based identification depends on the nucleic acid polymerase chain reaction (PCR), the quantity and quality/purity of DNA will contribute significantly to the species authentication. In the present study, different DNA extraction and purification methods (3 classical methods and one commercial kit) were compared to produce the better isolated DNA for PCR amplification. Additionally, different methods for the estimation of DNA concentration and purity which is essential for PCR amplification efficiency were also evaluated. The result showed that classical DNA extraction methods (based on TNES-Urea) yielded a higher amount of DNA (11.30-323.60 ng/g tissue) in comparison to commercial kit/Wizard Promega (5.70-83.45 ng/g tissue). Based on the purity of DNA extract (A260/280), classical DNA extraction method produced relatively similar on DNA quality to the commercial kit (1.79-2.12). Interestingly, all classical methods produced DNA with A260/280 ratio of more than 2.00 on the blue mussel, in contrast with commercial kit. The commercial kit also produced better quality of DNA compared to the classical methods, showing the higher efficiency in PCR amplification. NanoDrop is promising as cheap, robust and safe UV-spectrophotometer method for DNA quantification, as well as the purity evaluation.Keywords: seafood authenticity, DNA isolation, polymerase chain reaction, NanoDrop, Picogreen

Background Because of their specific and biologically relevant cargo, urine extracellular vesicles (EVs) constitute a valuable source of potential non-invasive biomarkers that could support the clinical decision-making to improve the management of prostate cancer (PCa) patients. Different EV isolation methods differ in terms of complexity and yield, conditioning, as consequence, the analytical result. Methods The aim of this study was to compare three different isolation methods for urine EVs: ultracentrifugation (UC), size exclusion chromatography (SEC), and a commercial kit (Exolute® Urine Kit). Urine samples were collected from 6 PCa patients and 4 healthy donors. After filtered through 0.22 µm filters, urine was divided in 3 equal volumes to perform EVs isolation with each of the three approaches. Isolated EVs were characterized by spectrophotometric protein quantification, nanoparticle tracking analysis, transmission electron microscopy, AlphaScreen Technology, and whole miRNA Transcriptome. Results Our results showed that UC and SEC provided better results in terms of EVs yield and purity than Exolute®, non-significant differences were observed in terms of EV-size. Interestingly, luminescent AlphaScreen assay demonstrated a significant enrichment of CD9 and CD63 positive microvesicles in SEC and UC methods compared with Exolute®. This heterogeneity was also demonstrated in terms of miRNA content indicating that the best correlation was observed between UC and SEC. Conclusions Our study highlights the importance of standardizing the urine EV isolation methods to guaranty the analytical reproducibility necessary for their implementation in a clinical setting.

Dengue is an important systemic viral infection that is caused by the dengue virus. Ribonucleic acid (RNA) from dengue NS1 positive samples, collected randomly during dengue epidemic from October 2016 to October 2017 at Chugtai Lab, was extracted for nucleic acid. Both the detection and serotyping of dengue samples were performed using real-time PCR on Rotor Gene Q. From the 70 NS1 positive samples, 57 (81.4%) samples were confirmed to be positive for dengue virus RNA, while the remaining 13 (18.6%) were negative. Serotype 1 (DEN-1) was verified among all samples by in-house assay and using commercial kit FTD (Fast Track Diagnostics) dengue differentiation; it was concluded that our in-house assay is in 100% concordance with commercial kit. Serotype 2 (DEN-2) and serotype 3 (DEN-3) have been documented in Pakistan since 1994. But recent detection of serotype 1 in Pakistan is indicative of more severe dengue haemorrhagic fever in future due to reinfection.

The pandemic caused by SARS-CoV-2 resulted in increasing demands for diagnostic tests, leading to a shortage of recommended testing materials and reagents. This study reports on the performance of self-sampled alternative swabbing material (ordinary Q-tips tested against flocked swab and rayon swab), of reagents for classical RNA extraction (phenol/guanidine-based protocol against a commercial kit), and of intercalating dye-based one-step quantitative reverse transcription real-time PCRs (RT-qPCR) compared against the gold standard hydrolysis probe-based assays for SARS-CoV-2 detection. The study found sampling with Q-tips, RNA extraction with classical protocol and intercalating dye-based RT-qPCR as a reliable and comparably sensitive strategy for detection of SARS-CoV-2 - particularly valuable in the current period with a resurgent and dramatic increase in SARS-CoV-2 infections and growing shortage of diagnostic materials as well for regions limited in resources.

Food fraud has been and still is a problem in the food industry. It is detectable by several approaches, such as high performance liquid chromatography (HPLC), chemometric assays, or DNA-based techniques, each with its own drawbacks. This work addresses one major drawback of DNA-based methods, in particular their sensitivity to inhibitors contained in particular matrices from which DNA is isolated. We tested five commercial kits and one in-house method characterized by different ways of sample homogenization and DNA capture and purification. Using these methods, DNA was isolated from 10 different fruit species commonly used in plant-based foodstuffs. The quality of the DNA was evaluated by UV-VIS spectrophotometry. Two types of qPCR assays were used for DNA quality testing: (i) Method specific for plant ITS2 region, (ii) methods specific for individual fruit species. Based mainly on the results of real-time PCR assays, we were able to find two column-based kits and one magnetic carrier-based kit, which consistently provided fruit DNA isolates of sufficient quality for PCR-based assays useful for routine analysis and identification of individual fruit species in food products.

BackgroundThe Corona Virus Disease 2019 (COVID-19), caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), has become a serious infectious disease affecting human health worldwide and rapidly declared a pandemic by WHO. Early, several RT-qPCR were designed by using only the first SARS-CoV-2 genome sequence.ObjectivesA few days later, when additional SARS-CoV-2 genome were retrieved, the kit GPS™ CoVID-19 dtec-RT-qPCR Test was designed to provide a highly specific detection method and commercially available worldwide. The kit was validated following criteria recommended by the UNE/EN ISO 17025:2005 and ISO/IEC 15189:2012.MethodsThe present study approached the in silico specificity of the GPS™ CoVID-19 dtec-RT-qPCR Test and RT-qPCR designs currently published. The empirical validation parameters specificity (inclusivity/exclusivity), quantitative phase analysis (10-106 copies), reliability (repeatability/reproducibility) and sensitivity (detection/quantification limits) were evaluated for a minimum of 10-15 assays. Diagnostic validation was achieved by two independent reference laboratories, the Instituto de Salud Carlos III (ISCIII), (Madrid, Spain) and the Public Health England (PHE; Colindale, London, UK).ResultsThe GPS™ RT-qPCR primers and probe showed the highest number of mismatches with the closet related non-SARS-CoV-2 coronavirus, including some indels. The kits passed all parameters of validation with strict acceptance criteria. Results from reference laboratories 100% correlated with these obtained by suing reference methods and received an evaluation with 100% of diagnostic sensitivity and specificity.ConclusionsThe GPS™ CoVID-19 dtec-RT-qPCR Test, available with full analytical and diagnostic validation, represents a case of efficient transfer of technology being successfully used since the pandemic was declared. The analysis suggested the GPS™ CoVID-19 dtec-RT-qPCR Test is the more exclusive by far.

This study evaluated the cost-effectiveness of platelet-rich plasma (PRP) added to usual care versus usual care alone in elderly patients with chronic diabetic foot ulcer (DFU) from the Spanish health care system perpective. A 6-state Markov model with 3-month cycles was used to estimate costs and outcomes of wound healing and risk of recurrences, infections, and amputations over 5 years. Three treatment strategies were compared: ( a) usual care plus PRP obtained with a commercial kit, ( b) usual care plus PRP obtained manually, and ( c) usual care. Data on effectiveness were taken from a recent meta-analysis. Outcomes and costs were discounted at 3% and resources were valued in 2018 euro. Compared with usual care, the PRP treatment with the manual method was more effective and less costly (dominant option), whereas the PRP treatment with the commercial kit was more effective but also more costly, with the incremental ratio being above the cost-effectiveness threshold (€57 916 per quality-adjusted life year). These results are sensitive to the price of PRP kits (a 20% discount would make the PRP treatment a cost-effective option) and effectiveness data, due to the heterogeneity of primary studies. In conclusion, PRP treatment for DFUs could be considered a cost-effective or even cost-saving alternative in Spain, depending on the method of obtaining the PRP. Despite the dominance of the manual method, its general use is limited to hospitals and specialized centers, whereas PRP kits could be used in primary care settings, but their prices should be negotiated by health authorities.

PCR-based methods are a key tool for the diagnosis of toxoplasmosis in immunocompromised patients. Laboratory-developed protocols lack standardization. This study aimed to assess the performances of a commercial kit for the detection of Toxoplasma DNA in different specimens drawn from immunocompromised patients. This multicentric retrospective study included 227 DNA specimens (157 blood specimens, 22 bronchoalveolar fluid [BALF] specimens, 39 cerebrospinal fluid [CSF] specimens, and 9 miscellaneous specimens) collected between 2010 and 2015 from 126 immunocompromised patients. The specimens were selected based on previous laboratory-developed quantitative PCR (qPCR) analyses targeting either the rep529 element or the B1 gene, and the results were classified as positive, negative, and "negative of interest," where the latter was defined as representing either the last specimen with a negative result before a positive one or the first with a negative result following a positive result(s). All specimens were secondary tested using the Bio-Evolution Toxoplasma DNA assay targeting the T. gondii rep529 element. We found a 95.6% concordance rate for qualitative results obtained with laboratory-developed qPCR techniques and the commercial kit. The rate reached 99.3% in comparisons of rep529-based laboratory-developed PCR methods and the commercial kit. The quantifications obtained with the commercial kit and the rep529 laboratory-developed PCRs were in very good agreement. Sensitivity and specificity of the commercial kit were calculated at 98.8% and 100%, respectively. The Bio-Evolution Toxoplasma DNA assay appears to be a valuable method for the detection of Toxoplasma DNA in blood, BALF, and CSF specimens from immunocompromised patients.

Introduction: Carbapenems are the drugs of choice for the treatment of many multidrug resistant hospital acquired infections. Resistance to carbapenems also is not uncommon and is increasingly being reported now-a-days. Though various tests like modified hodge test are available for the detection of carbapenemases, they lack sensitivity and specificity. Recently, the Carba NP test has been introduced for carbapenemase detection which has been approved and included in CLSI guidelines. Aim: To identify the prevalence of carbapenemase producing isolates and the utility of Carba NP in house as well as commercial RAPIDEC Carba NP test in the identification of these carbapenemases. Materials and Methods: All the carbapenem resistant organisms isolated during the study period from July 2018 to December 2018 were included in the study. A total of 91 isolates were identified during the study period which were further tested for carbapenemase production by both in House Carba NP test as well as commercially available RAPIDEC Carba NP (Biomerieux) as per the existing protocols. Twenty five carbapenem sensitive isolates were also tested. Klebsiella pneumoniae BAA ATCC1705 and BAA ATCC-1706 were used as controls. Results: Out of 91, carbapenem resistant strains tested, 72 were identified as carbapenemase producers. Out of them, Klebsiella pneumonia accounted for 59.7% of the total carbapenemase producing organisms followed by Pseudomonas aeruginosa (25%) and E.coli (6.9%) followed by Acinetobacter and Enterobacter which constituted less than 5%. Among enterobacteriaceae, 43 out of 59 carbapenem resistant Klebsiella were carbapenemase producers whereas all the 5 carbapenem resistant E.coli were positive for Carba NP test. Among Pseudomonas 18 out of 21 were carbapenemase producers. All the positive isolates by in-house Carba NP test were also positive by commercial RAPIDEC test and vice-versa and none of the 25 carbapenem sensitive strains tested were positive for Carba NP test by either method indicating 100% correlation between the two methods studied. Conclusion: Carba NP in house as well as commercial RAPIDEC Carba NP test were equally effective in the identification of the carbapenemase producing gram negative bacilli. Wide adaptability of these tests by various laboratories will help in the early identification of these potentially spreading carbapenemase producers, which in association with appropriate treatment and infection control practices will prevent the emergence of these strains and will decrease the mortality and morbidity.

OBJECTIVES To measure the effects of platelet-rich plasma (PRP) on hair diameter in Pakistani population. MATERIALS AND METHODS The study was conducted in a private setup in the adult patients undergoing PRP treatment. Baseline platelets count was noted for each patient. PRP was prepared using a commercial kit. Three PRP injections were administered in the scalp at 0, 1, and 3 months of interval. The hair caliber was measured in the mid-scalp area using a separate stencil for each patient. The diameter of 10 hair (1 cm sq) was measured randomly using the caliper (2 hair on each side of the square and 4 in the center). After 6 months of 3rd injection, the hair caliber was measured and the results were analyzed statistically using Mann-Whitney's test. RESULTS Seven men and 3 women were included in the study. The mean age of the patients was 24.2 years (26.3 years in males and 19.3 in females). The average platelets count was found to be 283,580/μL. The mean hair caliber was 37.3 μm (39.1 μm in men and 33.0 μm in women). At the end of the study, the mean hair caliber was found to be 52.3 μm (P

The banning of all mesh for prolapse by the FDA has failed to recognize that there is a major difference between mesh sheets applied behind the vagina for POP and repair of POP by shortening and reinforcing damaged suspensory ligaments, much as is used in the midurethral sling (MUS) which retains endorsement by the FDA. Mesh sheets interfere with the vaginal elasticity essential for function and may cause pain and massive urine loss postoperatively, in some cases; tapes far less so, as they have little contact with vagina and work by shortening and reinforcing damaged ligaments in the same way as the MUS operation. The main pathogenesis of POP is leaching out of collagen after the menopause. Weakened ligaments cannot support the organs so they prolapse. Only artificial collagenous neoligaments created by implanted tapes (as per the MUS) can create the new collagen required to repair these ligaments. The major advances developed over the past 30 years for treatment of POP using these ligament repair techniques has in one stroke been abolished. The treatment of POP has been set back 100 years. In their twilight years, many women will suffer the consequences of loose posterior ligaments, major prolapse, urge, nocturia, chronic pelvic pain, obstructed micturition defecation and fecal incontinence. The FDA needs to reconsider its ban, at least for tapes, which are supported by individual, multicentre and long-term studies.

Chagas disease, caused by Trypanosoma cruzi affects about 6-8 million people worldwide. Although transmission by triatomine insects has been controlled, other means of transmission maintain the infection. These forms of transmission are responsible for introducing Chagas disease in other non-endemic countries of the world. Thus, Chagas disease, nowadays is a worldwide health problem. In Brazil, acai pulp and sugarcane juice have been associated with Chagas disease outbreaks. The difficulties in isolation of the parasite from foods are hampering source tracking which could allow the confirmation of an implicated food commodity in these outbreak investigations. To address this scientific gap, we evaluated the performance of real-time PCR (qPCR) for detecting T. cruzi in acai pulp and sugarcane juice. All experiments were performed with acai pulp and sugarcane juice samples contaminated with different concentrations of T. cruzi. In assays with qPCR, the results showed that the ideal procedure for T. cruzi identification in acai pulp and sugarcane juice consisted of: i. centrifugation; ii. DNA extraction with a commercial kit for stool matrix; and iii. qPCR using a specific molecular marker for T. cruzi. The seeding in LIT medium of experimentally contaminated foods was effective in detecting the parasitic load by qPCR. The efficacy of qPCR was also verified testing food samples crushed with infected Triatomines. In conclusion, this methodology can be used to perform rapid diagnosis in outbreaks, facilitating measures in disease control.

SummaryBackground Postacne scarring is the main concern of many acne patients. Predicting the liability to scarring can alter the management protocols and help in preventing such disfigurement. Methods Sixty patients with moderate to severe acne vulgaris (according to Global Acne Grading system) were included, only 35 of them had postacne scars. A quantitative latex agglutination commercial kit was used to assess serum C-reactive protein (CRP) concentration by turbidimetry, while serum hepcidin concentration was assessed by a commercially available double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) kit. Results Patients with postacne scarring had significantly lower serum levels of hepcidin (P-value < .001) and significantly higher serum levels of CRP (P-value < .05). Conclusion Serum levels of hepcidin and CRP are promising markers, which may be considered as objective tools to predict the possibility of postacne scarring.

Objective: To assess the caries risk factors of preschool aged children. Methods: Data was collected from a sample of 85 preschool children and their mothers. An interview, and a clinical and microbiology assessment was included in this study. Clinical examinations were performed to measure the dental caries experience and plaque visibility of the children and mothers. The presence and levels of Mutans Streptococci (MS) in the saliva was measured using a commercial kit by Dentocult SM and evaluated using the manufacturer’s chart. Results: Sixty-five children had a high risk of caries (76.5%), 12.9% had a low risk, and 10.6% had a moderate risk of caries. Active cavities were found in more than half of the mothers and children. The majority of the child subjects had a snacking habit between meals (85.9%). High risk scores of 2 and 3, indicating salivary levels of MS, was found in 41.1% of children. Fluoride exposure experienced by the study population was very limited. Conclusion: Results suggest that the majority of the population had a high caries risk. There was a high occurrence of teeth with cavities in children and active carious lesions in the mothers. There was a high frequency in snacking habits, the presence of plaque and bleeding gums, high levels of salivary MS, and low protective factors for the children. Results suggest that management of caries related factors are needed in order to prevent caries in the future.

Background: The antibody against the transcriptional intermediate factor 1γ (Anti-TIF1γ or anti-p155), is considered specific for dermatomyositis (DM) and its a marker of cancer risk. Our center carries out the determination of anti-TIF1γ by a homemade immunoblot (IB) technique with a 155 kDa TIF1γ human recombinant protein (OriGene). Currently there is a commercial technique of IB to detect TIF1γ (Euroimmun), however, commercial kits sometimes elaborate antigens without using whole protein structure. To date, there are no studies that study the agreement between the commercial kit and homemades IB. Objectives: 1-To compare the commercial IB results of anti-TIF1γ with homemade IB. 2-To describe and compare the clinical characteristics of the patients analyzed by both techniques. METHODS: Observational retrospective study that included adult patients with some determination of anti-TIF1γ antibodies since 2014 in two tertiary-level university hospitals. We collected demographic and clinical data of all patients. Taking into account their diagnoses we grouped them into: Cancer associated Myositis (CAM), DM without cancer, Systemic autoimmune Diseases (SAD) non-DM and other diagnoses. Subsequently, the clinical agreement with the IB results of anti-TIF1γ performed by commercial (cIB) and homemade (hIB) technique were analyzed. A p value RESULTS: 48 patients were recruited (77.1% women). 13 had diagnosis of CAM, 19 of DM without cancer, 7 of other non-DM EAS, and 9 had other diagnoses. Of all DM (CAM and DM without cancer), 12 were positive for both techniques. 5 presented discordant results (4 hIB positive with negative cIB and 1 cIB positive with negative hIB). 15 DM were negative for the two techniques, of which 8/15 had other antibodies associated with autoimmune Myopathy (3 NXP-2, 1 Mi2, 1 MDA5, 1 SAE, 1 Jo1 and 1 Ro52). When we analyzed patients with CAM with both techniques, 10/13 presented positive hIB vs 8/13 cIB positive. DM without cancer, 6/19 had positive hIB vs 5/19 cIB positive. The sensitivity and specificity of each test was evaluated in CAM and DM without cancer, being 61.5% and 73.7% respectively for cIB (p = 0.071), compared to 76.9% and 68.4% for hIB (p = 0.029). In non-DM SAD group, 1 patient had positive determination for both techniques, 3 had positive cIB with negative hIB, and 3 cIB negative with positive hIB. We also found other antibodies in 6/7 patients with non-DM SAD. The 9 patients with other diagnoses were: 1 positive for both techniques and 8 positive for cIB and negative for hIB. CONCLUSIONS: Homemade IB with 155 kDa recombinant protein was superior to cIB to detect TIFγ in our serie of patients with CAM. This could have clinical implications in screening and early detection of neoplasia that affect false negative by cIB. A positive cIB in patients not suspected as a SAD were negative for hIB in the major part of cases. It would be recommended to check the commercial IB using homemade IB, due to its known clinical-therapeutic implications. Disclosure of interests: Ana Milena Millan arciniegas: None declared, M. ANGELES MARTINEZ: None declared, aNAIS MARISCAL: None declared, aNDRES BAUCELLS: None declared, Leticia alserawan: None declared, Ernesto Trallero: None declared, Cesar Diaz-Torne: None declared, ana Laiz Consultant for: Lilly, Novartis, abbVvie, MSD, UCB and Janssen, Speakers bureau: Lilly, Novartis, abvvie, MSD, UCB and Janssen, Berta Magallares: None declared, Patricia Moya: None declared, HyeSang Park: None declared, andrea Garcia-Guillen: None declared, Sycille Jeria: None declared, DAVID LOBO: None declared, albert Gil: None declared, Coral Gozalvez: None declared, Susana P. Fernandez-Sanchez: None declared, Laura Martinez-Martinez: None declared, Esther Moga: None declared, Hector Corominas: None declared, Candido Juarez: None declared, Ivan Castellvi Consultant for: I received fees less than 5000USD as a consultant for Kern and actelion, Paid instructor for: I received fees less than 2000USD as a instructor for Boehringer -Ingelheim, Novartis and Gebro, Speakers bureau: ND, albert Selva-O’Callaghan: None declared

Background Autoantibodies against apolipoprotein A1 (anti-apoA1 IgG) have emerged as an independent biomarker for cardiovascular disease and mortality. Across studies, different ELISA methods have been used to measure the level of circulating anti-apoA1 IgG which could lead to substantial result differences between assays. Objectives To make a comparative study of available anti-apoA1 IgG detection methods and to determine whether the choice of matrix sample (serum vs plasma) could influence the results. Methods Blood samples were obtained from 160 healthy blood donors and collected on 4 different matrixes (serum, plasma-EDTA, -citrate, -lithium-heparinate). Anti-apoA1 IgG was measured using two homemade (Geneva's and Lisbon's) and one commercial ELISA kits. Passing-Bablok and Bland-Altman were used to compare the results. Anti-apoA1 IgG seropositivity cut-offs were defined according to the user's/manufacturer's criterion. Results The current results showed substantial differences between those 3 assays. The dynamic ranges were significantly different, the commercial kit displaying the narrowest one. Passing-Bablok analysis demonstrated important proportional and constant biases between assays. The anti-apoA1 IgG seropositivity rate in Geneva, Lisbon and commercial assays varied between 24.5% and 1.9%. Matrix comparisons demonstrated that the matrix choice (plasma versus serum) influenced anti-apoA1 IgG results as well as the seropositivity rate in an assay-dependent manner. The coating antigen source was identified as important factor underlying results heterogeneity across assays. Conclusions These results highlight the impact of the method and the cut-off used on anti-apoA1 IgG results and emphasize the need of standardizing existing assays. Given the important matrix influence, we suggest to use serum as matrix of choice.

This study was aimed to assess the performance and suitability of commercial rapid test compared to the conventional test through verification process. This study used frozen meat from laboratory routine samples which divided into five groups, those were: natural group, low bacteria level group, medium bacteria level group, high bacteria level group, and control group, each sample test performed 9 replicates. All samples were tested for E. coli by conventional method (SNI 2897:2008) and commercial rapid test method. E. coli test result from both methods was calculated as precision (relative standard deviation), sensitivity, specificity, false negative, false positive, and kappa. The results showed that the commercial kit test had good precision with relative standard deviations score was 0.103. The sensitivity, specificity, false negative, false positive, and kappa score were 94.44%, 100%, 5.56%, 0%, and 0.872, respectively, indicates an equal performance with conventional method. The t student test showed that commercial rapid test method and conventional method had suitability on natural group, low bacteria levels group and medium bacteria level group.

Objective: Drinking water analysis is an important quality monitor and forms an integral part of the infection control policy of every hospital. On an average daily basis, 1750 liters of drinking water are used by patients and hospital staff. Presence of contaminating microorganisms in drinking water should therefore be identified at the earliest and necessary action be taken in order to prevent outbreaks through contaminated water. This study was conducted to compare two methods for drinking water analysis and document benefits of the new method. Methods: This cross sectional study was conducted over a period of two years at a tertiary care hospital in Chennai. A newer rapid method of drinking water analysis was evaluated for two years from July 2017 to June 2019. Parallel testing with conventional multiple tube method was performed. All samples collected from various sources during the study period were subjected to parallel testing by multiple tube method (conventional method for most probable number) and presence absence method (new method). Results: The new commercial kit for drinking water analysis comprehensively detects all water borne bacterial pathogens associated with gastroenteritis within 18 hours. The major advantage is marked reduction in turnaround time by 54 hours. False positive/false negative results were not encountered compared to the standard conventional method. Conclusion: Safer, healthier and high quality of water is ensured for use at an earlier and cost effective as well as environment friendly system. There was marked reduction in turnaround time, cost of testing, manpower usage, reagent wastage, waste generation using the kit based presence absence method. The new commercial kit can effectively replace the conventional method as an effective screening tool for drinking water analysis.

BACKGROUND The current common practice when using urine as a biomarker for vitamin excretion is to use a 24-hour sample for analysis. Due to the difficulty involved in this process, we attempted to find an alternative solution through the use of a single first morning void. The aim of our study was to investigate if there is a correlation between the first morning single void and the 24-hour collections of urines for the urine metabolite of niacin, N-1-methylnicotinamide (N1MN), and to test the reliability of utilizing a method using first morning single void collections corrected with the concentration of urine creatinine. MATERIAL AND METHODS All urine samples were collected from 30 healthy adult volunteers over the age of 18 years: 20 females and 10 males. Samples were collected after discarding the first morning urine and collecting every other urine voided during the next 24 hours including the first morning urine of the day after in 2 separate vessels. We analyzed the concentration of N1MN by high performance liquid chromatography and the concentration of creatinine by a commercial kit by spectrophotometry. The B3 excretion was expressed as the ratio of N1MN to creatinine. RESULTS We found a significant correlation between the ratios of first morning single void and 24-hour urines. When comparing males and females, the ratio demonstrated a significant correlation as well. CONCLUSIONS Our results demonstrated that it is possible to substitute a 24-hour collection with a first morning single void urine for the estimation of N1MN excretion.

Objective. The development of an accurate, sensitive, specific, rapid, reproducible, stable-at-room-temperature and cost-effective diagnostic kit, and a low-cost portable fluorescence detector to fulfil the requirements of diagnostic facilities in developing countries. Methods. We developed the ‘Chlamy and Ness CT/NG kit’ based on molecular beacons for the detection of Chlamydia trachomatis (CT) and Neisseriagonorrhoeae (NG). Multi-centric evaluation of the CT/NG kit was performed using the commercially available nucleic acid amplification test (NAAT)-based FTD Urethritis basic kit for comparison from December 2014 to November 2016. The stability of the kit reagents at 4 and 37 ˚C and the inter-day reproducibility of results were also analysed. Results. The sensitivity and specificity of the kit were found to be 95.83 and 100.00 % for the detection of C. trachomatis and 93.24 and 99.75 % for N. gonorrhoeae, respectively, when tested against the commercial kit. The positive predictive value (PPV) was 100.00 and 98.57 %, whereas the negative predictive value (NPV) was 99.54 and 98.79 % for C. trachomatis and N. gonorrhoeae, respectively. Analysis of the kappa statistics enhanced the ‘inter-rater’ κ=0.976 for Chlamydia and κ=0.943 for Neisseria. Conclusion. Our kit was found to be as sensitive and specific as commercially available kits. Its low cost and ease of use will make it suitable for the routine diagnosis of C. trachomatis and N. gonorrhoeae in the resource-limited settings of developing countries.

Acute respiratory infections caused by influenza virus are a major cause of viral respiratory diseases globally. Surveillance of circulating subtypes and estimation of disease burden is of utmost clinical importance. Molecular surveillance and proper disease burden estimates are scarce in India although clinical influenza infections are on the rise. Our study aims to delineate the prevalent influenza subtypes in a South Indian population from cases requiring hospital visits. Using real-time polymerase chain reaction (RT-PCR), 2154 throat/nasopharyngeal swabs from patients attending Government Medical College, Thiruvananthapuram, Kerala, India, with suspected influenza-like illness, were tested for the presence of different influenza subtypes. Forty-three percent of specimens were positive for the influenza virus. Among these, prevalence of influenza A(H3N2), influenza B, and H1N1pdm09 was 26.7, 6.3, and 10%, respectively. Nominal co-infections were detected. An easy to use commercial kit was used for the majority of the study after proper evaluation for sensitivity and specificity against a gold standard protocol. Specific diagnosis using molecular tools caters to the urgency, and a precise measure of the disease burden and management actions are needed, especially in developing countries like India. Infection rate estimation using a sensitive RT-PCR assay signified that influenza was highly prevalent in the region. The study data generated will help understand the epidemiology of influenza in India as well as generate information for global influenza surveillance and disease burden.

Background Diagnosis of Periplaneta americana (American cockroach, ACR) allergy is commonly performed based on clinical history and skin prick test (SPT) or specific serum IgE (sIgE) measurement. The concordance of the findings with the SPT and sIgE results has never been investigated. Objective To compare the results of SPT with commercial ACR-extract (C-ACE) and sIgE measurement, using commercial kit and in-house enzyme-linked immunosorbent assay (ELISA) to the locally produced ACR extract (L-ACE) and native Per a 1, Per a 5, Per a 7, and Per a 9. Methods Sera from 66 individuals clinically diagnosed with chronic allergic rhinitis were included; 46 were positive SPT to C-ACE, and 20 were negative. Specific serum IgE levels were established by using a commercial test kit (ImmunoCap) and an in-house IgE-ELISA Results The percentage the C-ACE SPT-positive cases that were positive by the ImmunoCap-sIgE was 32.6%, indicating low concordance of the 2 assays. With the in-house ELISA, Per a 9 gave the highest sensitivity (98.00%), positive predictive value (PPV; 95.74%), and negative predictive value (NPV; 94.74%) of the sIgE quantification. The correlation coefficients (R) of the L-ACE-SPT and sIgE to L-ACE, Per a 1, Per a 5, Per a 7, and Per a 9 and ImmunoCap sIgE were 0.133, 0.278, 0.419, 0.280, and 0.432, and 0.256, respectively. Conclusion Skin prick test and sIgE measurement using commercial reagents have low concordance. Data of this study showed that sIgE to the native Per a 9 should be considered as an adjunct to the clinical history in diagnosis of ACR sensitization/allergy, particularly when the SPT and the nasal challenge, which is the gold standard method, cannot be performed.

Given the multitude of potential sites of PRP treatment, it is important to understand the biological mechanism of its action, its indications, and its potential complications. The availability of commercial kits (each with its own instructions) makes harvesting easier for the physician, but it is still the physician’s responsibility to understand the benefits of different harvesting, preparation, and activation techniques to maximize the therapeutic effect. The basic equipment and protocol (if one chooses not to use a commercial kit) is also integral to the application of PRP therapy.

The microbiological standard method for the detection of Salmonella spp. is time-consuming, and consequently there is a need for an alternative rapid methodology for its detection. In this study, two open-formula real-time PCR methods for detecting the inv A and ttr gene Salmonella spp. have been successfully optimized and in-house validated. Different DNA extraction procedures were used (boiling, Chelex resin and standard I - Biorad). The relative sensitivity and false positive ratio for the alternative methods were 100% and 0.0%, respectively. The relative trueness was in range from 96.8% to 99.2%. No false negative results were detected. The lowest Ct values were obtained using the protocol for detecting the ttr gene after DNA extraction by the Chelex. The results were compared in a large number of food and environmental samples to those of the SRPS EN ISO 6579:2008 standard method and commercial kit (IQ check® Salmonella II kit, Bio-Rad, USA). Practical applications The whole procedure of real-time PCR methods in this study was approximately 20 hr, in contrast to 4–5 days of analysis time for the standard method (SRPS EN ISO 6579:2008). The real-time PCR method also provides a high level of sensitivity and specificity with a low risk of cross-contamination. The implementation of the method for routine analysis will help improve safety in the food production chain, by providing results compatible with the ISO standard, but more rapidly.

OBJECTIVE The aim of the present study was the evaluation of serology results in Mugla Sitki Kocman University Training and Research Hospital Microbiology Laboratory between 2012 and 2013 among cases from different divisions for anti-Toxoplasma antibodies, retrospectively. METHODS Anti-Toxoplasma IgG and IgM antibodies were detected by a commercial kit in serum samples. The age and gender of cases, the division where the samples were sent and the changes in the seropositivity between years were evaluated statistically by chi-square test. RESULTS Totally 1162 cases varying from 9 months to 85 ages (mean 29.1 ± 10.8) were included in the study, due to the orders from divisions anti-Toxoplasma IgG antibodies were investigated in 747 samples and 154 (20.6%) of them were positive. Anti-Toxoplasma IgM antibodies were investigated in 1112 samples and 27 of them (2.4%) were positive. The seropositivity of anti-Toxoplasma IgG antibodies were increased with age (p = 0.001) and a significant relationship was found with anti-Toxoplasma IgM seropositivity and the division where the samples were sent (p = 0.003). CONCLUSION It can be concluded that as in the other parts, toxoplasmosis is an important public health concern in Mugla and it is crucial especially for women in reproductive age, the risk group for toxoplasmosis, should be informed about toxoplasmosis.

The objective of this study was to evaluate performance of the commercial kit based on loop-mediated isothermal amplification (LAMP) in comparison with the International Organization for Standardization method for detecting uninjured and sublethally injured Salmonella cells artificially inoculated at levels of 10(0) and 10(1) CFU/25 g on raw duck wing, raw mung bean sprouts, and processed fishballs. Injured cells were prepared by a heat treatment for duck wings and fishball samples and a chlorine treatment for bean sprout samples. Additionally, a validation study was performed on naturally contaminated food samples sold in Singapore. A total of 110 samples of each commodity were analyzed in this study. Regardless of inoculum levels, the detection by the commercial LAMP kit showed 100% sensitivity and specificity for both inoculated and uninoculated samples compared with the International Organization for Standardization method, with the exception of bean sprout samples. Only 20% of bean sprout samples inoculated with 10(0) CFU/25 g injured Salmonella cells were positive by using the commercial LAMP-based kit. However, all negative samples became positive following a secondary enrichment in Rappaport-Vassiliadis medium with soy broth or after concentration by centrifugation. These results suggest that secondary enrichment or centrifugation should be considered as an additional step to increase the sensitivity of the commercial LAMP-based kit with low numbers of injured target cells in samples with high background microflora (such as mung bean sprouts). The validation study also showed that the commercial LAMP-based kit provided 91% sensitivity and 95% specificity for naturally contaminated samples. Thus, this study demonstrates that the commercial LAMP-based kit might be a cost-effective method, as this system could provide rapid, accurate detection of both uninjured and injured Salmonella cells on raw duck wings, raw mung bean sprouts, and processed fishballs in less than 26 h.

The aim of the present study was to compare two protocols of Agar Gel Immunodiffusion (AGID) that can be used to diagnose Equine Infectious Anemia (EIA). Different volumes of reagents and support for agar gel were used, as recommended by the Ministry of Agriculture, Livestock and Food Supply (MAPA) and the World Organization for Animal Health (OIE). Equidae sera samples (n = 1000) were tested using a national commercial kit and different volumes (i.e. 25 μL—MAPA and 50 μL—OIE) of reagents and standard sera. Antigen was used as recommended by MAPA. The results were assessed by two operators, 48 hours after beginning the test. At the end of the experiment, a Kappa index of 1 was obtained by the operators, for the interpretation of the results and for the different volumes of reagents used. Similar results of agreement (Kappa index = 1) were obtained when using slides or Petri dishes as support for the agar gel. In both cases, an almost perfect agreement was achieved, suggesting that the use of different supports does not affect the results. These findings indicate that both protocols (MAPA and OIE) can be used to diagnose EIA.

SETTING: A tertiary care hospital in North India. OBJECTIVE: To evaluate a commercial kit-based loop-mediated isothermal amplification (TB-LAMP) assay for the diagnosis of pulmonary tuberculosis (PTB). DESIGN: A total of 530 patients presenting with PTB symptoms were enrolled and one sputum sample was collected from each patient. The TB-LAMP assay (Loopamp™ MTBC Detection kit) was performed on the raw sputum sample. The remaining sample was used for smear microscopy and mycobacterial culture. A cartridge-based nucleic acid amplification test (NAAT, Xpert® MTB/RIF assay) was also performed on the processed pellet. RESULTS: The sensitivity and specificity of the TB-LAMP assay in culture-positive samples obtained from 453 patients presenting with PTB symptoms (77 specimens were excluded) were respectively 100% (95%CI 94.7-100) and 99.2% (95%CI 97.8-99.8). The sensitivity and specificity of Xpert in culture-positive samples were respectively 82.6% (95%CI 71.5-90.6) and 94.9% (95%CI 92.2-96.8). A concordance of 0.75 was obtained between the two NAATs (TB-LAMP assay and Xpert) using the κ statistic. CONCLUSION: The TB-LAMP assay showed high sensitivity and specificity with limited requirement of testing infrastructure, and is thus a promising diagnostic tool for TB diagnosis in resource-poor settings.

BACKGROUND Measurement of viral load in human immunodeficiency virus type 1 (HIV-1) infected patients is essential for the establishment of a therapeutic strategy. Several assays based on qPCR are available for the measurement of viral load; they differ in sample volume, technology applied, target gene, sensitivity and dynamic range. The Bioneer AccuPower® HIV-1 Quantitative RT-PCR is a novel commercial kit that has not been evaluated for its performance. OBJECTIVE This study aimed to evaluate the performance of the Bioneer AccuPower® HIV-1 Quantitative RT-PCR kit. METHODS In total, 288 EDTA plasma samples from the Dharmais Cancer Hospital were analyzed with the Bioneer AccuPower® HIV-1 Quantitative RT-PCR kit and the Roche COBAS® AmpliPrep/COBAS® TaqMan® HIV-1 version 2.0 (CAP/CTM v2.0). The performance of the Bioneer assay was then evaluated against the Roche CAP/CTM v2.0. RESULTS Overall, there was good agreement between the two assays. The Bioneer assay showed significant linear correlation with CAP/CTM v2.0 (R2=0.963, p

This study was supported by the Minas Gerais State Research Support Foundation (FAPEMIG) protocol number 65/10 and the National Council for Scientific and Technological Development [Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq)] protocol number 310174/2014-7-CNPQ. Universidade Federal de Minas Gerais. Faculdade de Medicina. Grupo de Pesquisa em Micobacterioses. Belo Horizonte, MG, Brasil. Universidade Federal de Minas Gerais. Faculdade de Medicina Grupo de Pesquisa em Micobact?rias. Belo Horizonte, MG, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Federal de Minas Gerais. Faculdade de Medicina Grupo de Pesquisa em Micobact?rias. Belo Horizonte, MG, Brasil. Universidade Federal de Minas Gerais. Faculdade de Medicina Grupo de Pesquisa em Micobact?rias. Belo Horizonte, MG, Brasil. Universidade Federal do Rio de Janeiro. Programa Acad?mico de Tuberculose. Rio de Janeiro, RJ, Brasil. Universidade Federal de Minas Gerais. Faculdade de Farm?cia. Belo Horizonte, MG, Brasil. Universidade Federal de Minas Gerais. Consultoria e Apoio T?cnico. Belo Horizonte, MG, Brasil. Introduction: This study aimed to evaluate a new commercial kit, Kit SIRE Nitratase-PlastLabor, for testing the drug susceptibility of clinical Mycobacterium tuberculosis isolates. Methods: The accuracy of the Kit SIRE Nitratase was evaluated by examining the susceptibility (streptomycin, isoniazid, rifampicin, and ethambutol) of 40 M. tuberculosis isolates, using the proportion method with Lowenstein-Jensen medium or the BACTEC MGIT 960 system. Results: The detection accuracy for streptomycin, isoniazid, rifampicin, and ethambutol was 95%, 97.5%, 100%, and 80%, respectively. Conclusions: The exceptional accuracy demonstrated by Kit SIRE Nitratase for isoniazid and rifampicin makes the kit an attractive option for screening M. tuberculosis strain resistance.

Introduction Accurate evaluation of ADAMTS13 activity is required for the diagnosis and clinical management of thrombotic microangiopathies, and commercial kits are available for routine laboratory use. Methods Our study compares the results from Technoclone (Technoclone GmbH, Austria) activity and Inhibitor kits with specialist laboratory reference methods (FRETS and ELISA IgG) and the impact of transporting frozen samples and comparison of results. Results This multicentre study identified differences in Technoclone activity results compared to specialist testing, which could potentially impact diagnosis. A change in the commercial kit during the study period appears to have rectified the detection levels. Frozen samples provided comparable results between sites. Conclusion With close attention to normal ranges, commercial kits are suitable for use in the clinical diagnosis of thrombotic microangiopathies and frozen transportation of samples between sites is a suitable approach. However, a robust external quality control system is essential to provide an independent evaluation of changes in kit production.

Background. Autologous Conditioned Plasma (ACP) has a wide range of potential uses in modern orthopaedics. The aim of this study was to examine the characteristics of proprietary ACP and compare them with those of ACP produced using a commercially available kit. Material and methods. In the hospital laboratory, 20 samples of ACP taken from patients and prepared according to the commercially available kit protocol with a double syringe system were compared with 40 ACP preparations made using disposable sterile equipment available in the hospital. Results. The mean platelet concentration in the ACP samples prepared according to the Arthrex protocol was 2.02 (range 1.16 to 2.64) times greater than in peripheral blood, while the concentration in the proprietary preparation was 1.61 (range 0.82 to 2.52) times higher. However, the mean platelet density in the proprietary preparation (n = 24) was 1.98 (range 1.48 to 2.52) times that of peripheral blood within 20 minutes of collection, and 1.41 (range 0.84 - 1.87) times after 20 minutes (n = 16). Therefore, the proprietary method of producing ACP is comparable to that of the commercial kit with regard to platelet density (p>0.05). Conclusion. Using disposable hospital equipment and with a relatively short time between ACP preparation and testing, it is possible to obtain a suitable proprietary platelet-rich preparation comparable to one produced using a commercial system.

Background:Recently, the total prostate-specific antigen (PSA) assay used in a laboratory had a positive bias of 0.03 μg/L, which went undetected. Consequently, a number of post-prostatectomy patients with previously undetectable PSA concentrations (defined as Methods:Through numerical simulations, we explored (1) how a small bias may evade the detection of routine quality control (QC) procedures with specific reference to the concentration of the QC material, (2) whether the use of ‘average of normals’ approach may detect such a small bias, and (3) describe the use of moving sum of number of patient results with detectable PSA as an adjunct QC procedure.Results:The lowest QC level (0.86 μg/L) available from a commercial kit had poor probability (Conclusions:Laboratory practitioners should ensure their QC strategy can detect small but critical bias, and may require supplementation of ultra-low QC levels that are not covered by commercial kits with in-house preparations. The use of moving sum of number of patient results with a detectable result is a helpful adjunct QC tool.

Food allergy is recognised as an important human health problem. Fish represent one of the most important causes of food hypersensitivity reaction. Small amounts of the allergen can cause severe reactions in sensitive individuals, so correct labelling is essential to ensure the protection of consumers. The objective of the present work was to develop a reliable, sensitive and specific real-time PCR method for the detection of fish and traces of fish in all kind of products included those that have undergone aggressive treatments such as high temperature or pressure. This methodology was validated simulating products likely to contain this allergen and spiking them with fish cooking water. In addition, a comparison between the performance of in-house methodology and a commercial kit, both of them based on real-time PCR, was carried out. This work is relevant because it is the first, rapid real-time PCR method developed to date for the detection of fish in processed food products. The results obtained confirm the present assay is a useful tool in detecting fish and, therefore, minimising exposure and reducing incidences of allergic reaction to fish in contaminated products.

Molecular markers are important for characterizing the genetic diversity of plants and can provide the basis for strategies to protect and conserve endangered populations. However, numerous molecular techniques are used, requiring an evaluation of fast and efficient methods to extract DNA. Since molecular studies of Caesalpinia ferrea are rare, it is important to develop and/or adapt a DNA extraction protocol that produces quality DNA samples to enable the design of strategies for the conservation of this threatened species. This study aimed to compare five methods for DNA extraction and to determine the most efficient protocol for C. ferrea. Sufficient genomic DNA was obtained from the leaves of C. ferrea using all the tested protocols to perform techniques involving molecular markers. Two protocols based on the detergent cetyl trimethyl ammonium bromide, as well as a commercial kit, yielded high concentrations of pure DNA. However, when polymerase chain reaction amplifications were performed, DNA was only successfully amplified from extractions performed with the commercial kit, which produced sufficient genomic DNA of good quality from the leaves of C. ferrea to perform techniques involving molecular markers.

Human noroviruses (HuNoVs), a leading cause of food-borne gastroenteritis worldwide, are easily transferred via ready-to-eat (RTE) foods, often prepared by infected food handlers. In this study, the transmission of HuNoV and murine norovirus (MuNoV) from virus-contaminated hands to latex gloves during gloving, as well as from virus-contaminated donor surfaces to recipient surfaces after simulated preparation of cucumber sandwiches, was inspected. Virus transfer was investigated by swabbing with polyester swabs, followed by nucleic acid extraction from the swabs with a commercial kit and quantitative reverse transcription-PCR. During gloving, transfer of MuNoV dried on the hand was observed 10/12 times. HuNoV, dried on latex gloves, was disseminated to clean pairs of gloves 10/12 times, whereas HuNoV without drying was disseminated 11/12 times. In the sandwich-preparing simulation, both viruses were transferred repeatedly to the first recipient surface (left hand, cucumber, and knife) during the preparation. Both MuNoV and HuNoV were transferred more efficiently from latex gloves to cucumbers (1.2% ± 0.6% and 1.5% ± 1.9%) than vice versa (0.7% ± 0.5% and 0.5% ± 0.4%). We estimated that transfer of at least one infective HuNoV from contaminated hands to the sandwich prepared was likely to occur if the hands of the food handler contained 3 log 10 or more HuNoVs before gloving. Virus-contaminated gloves were estimated to transfer HuNoV to the food servings more efficiently than a single contaminated cucumber during handling. Our results indicate that virus-free food ingredients and good hand hygiene are needed to prevent HuNoV contamination of RTE foods.

Objective: To establish a time-resolved fluoroimmunoassay (TRFIA) method to detect p16INK4a. Methods: 119 cases of exfoliative cells following Thinprep cytologic test (TCT) test were selected for TRFIA and biopsy tissues of these patients according to TCT results were collected for immunohistochemical (IHC) staining. Results: There were 40 cases with cervicitis, 16 cases of cervical intraepithelial neoplasia grade 1 (CIN1), 15 cases of CIN2, 26 cases of CIN3, and 22 cases of squamous cell carcinoma (SCC). For each group, TCT results displaying above ASC-US were 7, 6, 12, 23, and 21 cases. IHC results were positive for 2, 13, 14, 26, and 21 cases, while the TRFIA results were positive for 5, 12, 13, 25, and 21 cases, respectively. The sensitivity and specificity were not significantly different between TCT and TRFIA test due to the limited cases. Significantly more CIN1 cases were positive by TRFIA, as compared with cervicitis (75.00% vs. 12.50%, χ 2 =21.116, p 2 =4.877 and p =0.027). Conclusion: Detecting p16INK4a protein by TRFIA is suitable to discriminate cervical dysplastic/neoplastic lesions from cervicitis. A commercial kit which created in the future might be a rapid, simple, and low-cost method to obtain objective results with an instrument automatically on a large scale.

Objective. To study the frequency of identifying DNA of human herpesvirus type 4 (Epstein-Barr virus – EBV) in different brain tumours. Materials and methods. Following the surgical resection, 89 samples of brain tumours were analyzed by real time PCR (“DNA technology” commercial kit) for the presence of EBV DNA. Results. EBV DNA was detected in about 40% of the samples of different brain tumours. The frequency of EBV DNA in the samples of extracerebral tumours was higher than in intracerebral glial tumours. EBV DNA positivity in benign gliomas was superior to that in malignant ones. The mechanisms of EBV infection of tumour cells in the brain and the role of EBV in stimulation of tumour growth have not been yet elucidated. Conclusion. EBV DNA is detected in the cells of brain tumours of different histology, prevalently in glial tumours.

A survey of aflatoxin M1 (AFM1) contamination in packaged milk and infant formula milk samples in the Goan market, India, was conducted using high performance liquid chromatography, association of analytical communities approved commercial kit and a sensitive chemiluminescent sandwich enzyme-linked immunosorbent assay (ELISA). A total of 72 samples of infant formula milk food (18) and packaged milk samples (54) was analysed. One hundred per cent of the analysed samples exceeded the European Communities recommended limits (50 ng/L) and 75% of the samples exceeded Codex Alimentarius, Food Safety and Standards Authority of India (FSSAI) and US Food and Drug Administration recommended limits (500 ng/L). The range of contamination of AFM1 was found lower in infant milk formula (501–713 ng/L) than liquid milk (511–809 ng/L). The methods were also compared for their performance, and ELISA was found to be most suitable for analysis of low-level AFM1 contamination in milk.