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Among the strategies to tackle the COVID-19, much attention is paid to “test and isolation” advocated by Romer (2000), Peto et al (2020) etc., in addition to vaccination and development of medicines to treat COVID-19. According to these articles, what is necessary is a targeted version of lockdown, that is, to test everyone regularly and isolate the small fraction of the population who test positive. There is, however, concern that the PCR test is not always effective, so that, in the present paper, we investigate the effects of improvement of the PCR test by constructing a simple intertemporal theoretical model. Main result we obtain is that improvement of the PCR test could increase the number of the infected individuals at first, so that we should improve the accuracy of the PCR test much enough to reduce the number of the infected individuals.

The paper presents comparative test results of 12 domestically produced diagnostic kits/PCR test systems for DNA detection of the African swine fever virus with regard to the following parameters: completeness and correctness of instructions for use; labeling and package contents; convenience of using the kit; shelf life stability of reagents; stability of reagents after transportation and repeated freezing – thawing; batch-to-batch repeatability; sensitivity of various test materials and specificity of kits. The study of the instructions for use and kit contents revealed incompleteness of some instructions. It was noted that some manufacturers make serious errors in the instructions, which can significantly affect the interpretation of test results. It was also observed that there is insufficient control of the manufacturing process, which results in the production of faulty kits, as well as kits with poor-quality components and errors in the labeling. Thus, during the study, one kit showed its inactivity, demonstrating the absence of accumulation curves of the fluorescent signal during amplification of both positive controls and DNA of ASFV isolates. When the specificity was assessed, all the kits showed absence of non-specific reactions and acceptable sensitivity when testing various types of ASFV-containing material (blood, suspensions of pork spleen and pork casings used in sausage production). The stability test showed a sharp deterioration in the quality of operation of one kit within the shelf life period, and a significant decrease in the fluorescence signal was detected during repeated freeze – thaw cycles for another kit. Comparison of the repeatability results of different kit batches of the same manufacturer showed significant discrepancies for 41.5% of all kits. It was found that only 33% of the studied kits for ASFV DNA detection were compliant. The results of this study demonstrate the need for control of the manufactured diagnostic kits used in state programs for animal disease monitoring.

In Japan, the policy for polymerase chain reaction (hereafter PCR) testing changed significantly after 7 May 2020; from 4 February to 6 May, PCR testing was limited to certain patients with severe symptoms. After 7 May, the PCR test was made available to a broader range of patients due to health insurance coverage. The study aims to test whether there is a significant relationship between the conditions under which PCR tests are performed, the number of tests after 7 May, and the positive results. Using a multiple regression model, we obtained the unexpected result even if we assume that PCR testing had been carried out during 4 February to 6 May at the same level as after 7 May. The number of positive cases would have been even lower than the actual number, which we have attained. This suggests that even if PCR testing had been plentiful throughout the entire period, the number of positives that would have been captured would not necessarily have been more significant than the actual number. This estimation might suggest that the infectivity of COVID-19 varied over time. It may suggest that, over time, the infectiousness and spreading power of COVID may be transformed. Therefore, further research investigating the epidemic impact of COVID is required, which is critical for humankind.

Background The Simultaneous Multiplex Real Time PCR (SIMUL-qPCR) Listeria species and monocytogenes Assay is a quick, reliable method for detecting Listeria species and monocytogenes in environmental and food samples. The assay multiplexes several targets in one run to properly identify Listeria species and monocytogenes. The assay uses proprietary medium, Listeria Recovery and Enrichment Broth (LREB), for enrichment purposes. LREB was specifically formulated to improve the recovery and growth of Listeria while inhibiting competing background flora. Objective This report details the method validation study to validate frankfurters, ready-to-eat (RTE) sliced turkey, soft fresh raw cheese, chicken salad, ice cream, cooked eggs, pasteurized milk, and frozen/cooked shrimp, as well as environmental surface sponges and swabs for stainless steel, plastic, rubber, ceramic tile, and sealed concrete. Method Matrix studies, inclusivity/exclusivity, product consistency/stability, and robustness testing were conducted to assess the method’s performance. Results There were no statistically significant differences found between the candidate and reference methods in the matrix studies. Inclusivity/exclusivity testing showed that the assay was able to detect both Listeria species and monocytogenes strains while excluding the non-Listeria isolates. Small variations in critical test parameters (enrichment time, extraction reagent volume, and extracted sample volume) did not adversely affect the assay’s performance, and stability testing indicated consistent results for at least 1 year. Conclusions The data presented in this report show that this a reliable method for detecting Listeria species and monocytogenes. Highlights This assay allows for one sample to be tested for both Listeria species and monocytogenes with one PCR test.

Objectives We aimed to compare the efficiency of the CapitalBioMycobacterium real-time polymerase chain reaction (PCR) detection test with the standard Xpert MTB/RIF assay for the diagnosis of tuberculous meningitis (TBM). Methods We analyzed cerebrospinal fluid (CSF) from 163 patients with suspected TBM that were collected between January 1, 2018, and December 31, 2019. For both tests, we determined the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and area under the curve (AUC). Next, we compared the diagnostic accuracy of the two techniques using clinical diagnosis as a reference standard. Results The sensitivity, specificity, PPV, NPV, and AUC, of the CapitalBio Mycobacterium detection test were 48.5%, 100%, 100%, 29.6%, and 0.74, respectively, when used for the diagnosis of TBM. In comparison, the Xpert MTB/RIF assay returned values of 47.0%, 100%, 100%, 29.0%, and 0.74, respectively. Our analysis showed that the diagnostic accuracies of the CapitalBio Mycobacterium detection test and the Xpert MTB/RIF assay were very similar; the accuracy of both tests for detecting mycobacteria was significantly higher than that associated with acid­fast staining. Conclusions The CapitalBio Mycobacterium real-time PCR detection test has moderate sensitivity and very high specificity for TBM; results are very similar to those generated by the Xpert MTB/RIF assay. We recommend that the CapitalBio PCR test should be used as an initial screening method for TB.

Objective To describe the frequency and timing of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody detection in a convenience sample of skilled nursing facility (SNF) residents with and without confirmed SARS-CoV-2 infection. Design Retrospective analysis of SNF electronic health records. Setting Qualitative SARS-CoV-2 antibody test results were available from 81 SNFs in 16 states. Participants 669 SNF residents who underwent both polymerase chain reaction (PCR) and antibody testing for SARS-CoV-2. Measurements Presence of SARS-CoV-2 antibodies following the first positive PCR test for confirmed cases, or first PCR test for non-cases. Results Among 397 residents with PCR-confirmed infection, antibodies were detected in 4 of 7 (57.1%) tested within 7-14 days of their first positive PCR test; in 44 of 47 (93.6%) tested within 15-30 days; in 182 of 219 (83.1%) tested within 31-60 days; and in 110 of 124 (88.7%) tested after 60 days. Among 272 PCR negative residents, antibodies were detected in 2 of 9 (22.2%) tested within 7-14 days of their first PCR test; in 41 of 81 (50.6%) tested within 15-30 days; in 65 of 148 (43.9%) tested within 31-60 days; and in 9 of 34 (26.5%) tested after 60 days. No significant differences in baseline resident characteristics or symptoms were observed between those with vs. without antibodies. Conclusions These findings suggest that vulnerable older adults can mount an antibody response to SARS-CoV-2, and that antibodies are most likely to be detected within 15-30 days of diagnosis. That antibodies were detected in a large proportion of residents with no confirmed SARS-CoV-2 infection highlights the complexity of identifying who is infected in real time. Frequent surveillance and diagnostic testing based on low thresholds of clinical suspicion for symptoms and/or exposure will remain critical to inform strategies designed to mitigate outbreaks in SNFs while community SARS-CoV-2 prevalence remains high. This article is protected by copyright. All rights reserved.

Background Pneumocystis pneumonia (PCP) is a disease caused by the opportunistic infection of the fungus Pneumocystis jirovecii. Several PCR methods have been developed to aid in the diagnosis of PCP. In this study, we evaluated the performance of a real-time PCR in the diagnosis of PCP, in patients with various underlying diseases. Methods Ninety-seven BAL samples and 94 sputum samples from 191 patients were used in the study. Patients were classified as PCP (121 patients) or non-PCP (70 patients) based on their clinical and radiological presentations. Results Real time PCR amplified the P. jirovecii mitochondrial large-subunit rRNA gene with a detection limit of 68 copies of DNA per reaction. Non-PCP pathogens including 32 different fungi and bacteria were also evaluated. Overall, 71.9% of the samples from PCP patients and 14.5% of those from non-PCP patients were positive for the PCR test with a CT value of the real-time PCR below 45. The main underlying diseases of the patients were hematological or solid malignancies (47.1%) and HIV infection (8.9%). The CT values of the test were significantly lower in BAL samples from PCP patients than those from non-PCP patients (p = 0.024). No non-PCP patient had a CT value below 30, whereas samples from 24.8% of PCP patients with underlying diseases had a CT value below 30. Conclusion Since false positive PCR results were obtained, perhaps due to colonization, we suggest that the diagnosis of PCP should be based on a combination of clinical symptoms, underlying diseases, and PCR results.

Malaria shows various clinical manifestations from mild fever to death depending on the Plasmodium species. Among the complications, reports of malaria-associated splenic infarctions are rare. Here we present a case in which a man suffered from malaria-induced splenic infarction with serial follow-up. A 28-year-old man who served in the military near the Korean Demilitarized Zone (DMZ) was referred to the emergency room for fever beginning 1 week ago. He suffered upper abdominal pain for 2 days. At the time of his visit, he experienced a fever spiking up to 39.8°C. In the patient’s computed tomography (CT) test, splenomegaly with low attenuation density suggesting splenic infarction and hepatomegaly was shown. Because red blood cells infected by a plasmodium species were shown in a peripheral blood smear, he was admitted for malaria infection. The patient was given oral chloroquine on the day of admission and on hospital day (HOD) 3, Plasmodium vivax was detected in his malaria PCR test. After conservative management, the patient’s condition improved. The patient was discharged on HOD 15 without any symptoms. At this time, the patient’s spleen size decreased to the upper limit size of normal according to an ultrasonography. After that, the patient visited the outpatient department. Although low attenuation density still appeared in the following CT on HOD 30, a subsequent ultrasonography on HOD 60 did not show any specific finding. Although malaria-induced splenic infarction is still rare, this rate may increase. Most of the cases can be treated without surgery.

Objective Infection by Primate erythroparvovirus 1, generally known Parvovirus B19, is highly prevalent worldwide. Although infection by this virus will not be clinically problematic in most cases, new infections during pregnancy could result in serious repercussions in the fetus. Serologic and PCR-based methods are among the available approaches for diagnosis of Parvovirus B19 infection. In this regard, the present study is aimed to investigate the frequency of Parvovirus B19 infection by these two techniques in pregnant women of Zanjan. Materials and methods In this cross sectional-descriptive study, 110 pregnant women referring to Mousavi hospital in Zanjan during one year were evaluated in terms of serologic and Real-Time PCR test results in search for Parvovirus B19 infection. The rate of positive IgG and IgM were determined in women and the Real-Time PCR results were reported. Results Overall, 18.2% of participants were above 35 years old and 4.5% of them were younger than 18 years old. 41 (44.1%) and 2 (1.8%) cases had positive anti-Parvovirus B19 IgG and IgM, respectively. Real-Time PCR results were negative in all the studied samples. Conclusion Based on the findings of this study, prevalence of acute Parvovirus B19 infection was 0 and 2% based on Real-Time PCR and IgM tests, respectively. About 40% of pregnant women had experienced infection with this virus before.

This study carried out in Salahaldeen province in period January to September 2019. The aims of this study were to investigate the incidence of salmonella typhimurium and salmonella dublin in cow by using of PCR test. The results of current study showed that salmonella species isolation from cow in rate of 13.3% by culture methods, high of them from aborted cows in rate of 22.2%. Result of PCR test showed that salmonella typhimurium and salmonella Dublin detection in rate of 55% and 25% respectively while other salmonella species detected in rate of 20% from total salmonella isolates . Introduction Salmonellosis is one of most important zoonotic dangerous diseases, caused by bacteria return to genera of salmonella (1). Salmonella is gram negative bacilli, non-spore forming , non-capsulated, motile by Peritrichous Flagella except S. gallinarum and S. pullorum . lactose and sucrose non ferment while ferment of glucose, maltose and sorbetoil (2).

The impact of the COVID-19 pandemic in sport has been the subject of numerous studies over the past two years. However, knowledge about the direct impact of COVID-19 infection on the performance of athletes is limited, and the importance of studies on this topic is crucial during the current pandemic era. This study aimed to evaluate the changes in the match running performance (MRP) of professional soccer players that occurred as a result of COVID-19 infection after fulfilling all of the prerequisites for a safe return to play (RTP). The participants were 47 professional soccer players from a team which competed in first Croatian division (21.6 years old on average) during the 2020/21 season. The total sample was divided into two subgroups based on the results of a PCR test for COVID-19, where 31 players tested positive (infected) and 16 tested negative. We observed the PCR test results (positive vs. negative PCR), the number of days needed to return to the team, number of days needed to RTP after quarantine and isolation, and MRP (10 variables measured by a global positioning system). The number of days where the infected players were not included in the team ranged from 7 to 51 (Median: 12). Significant pre- to post-COVID differences in MRP for infected players were only found for high-intensity accelerations and high-intensity decelerations (t-test = 2.11 and 2.13, respectively; p < 0.05, moderate effect size differences), with poorer performance in the post-COVID period. Since a decrease of the MRP as a result of COVID-19 infection was only noted in two variables, we can highlight appropriateness of the applied RTP. However, further adaptations and improvements of the RTP are needed with regard to high-intensity activities., peer-reviewed

Introduction COVID19 has raised concerns for resource allocation across various sectors of healthcare. At the frontlines, emergency departments are required to triage a wide range of acuity and non-specific symptomology. Methods This retrospective study aimed to pave the way for more concrete detection and triage of patients by analyzing symptomology, physical findings, diagnostic testing and relevant hospital course of the 458 suspected cases that initially presented to an academic level one trauma center emergency department between March and August 2020. A total of 202 COVID positive cases were analyzed. Results The most common symptoms were cough (70.63%), fatigue (77%), and shortness of breath (59%). There was a significantly higher percentage of abnormal chest imaging in inpatient groups compared to the ED discharge group (42.86% vs 79%, p

Considering the increasing prevalence and burden of coronavirus disease 2019 (COVID-19) disease and false-negative results in routine reverse transcription-polymerase chain reaction (RT-PCR) tests, additional diagnostic methods are needed to diagnose active cases of this disease. This prospective study was conducted on patients, in whom clinical and radiological symptoms/signs were in favor of COVID-19 while their first PCR test was negative. Later on, a second RT-PCR was performed and serological evaluation was carried out and results were compared with each other. Out of 707 patients who had been referred to the hospital and were clinically and radiologically suspicious of disease, 137 patients with negative RT-PCR tests entered the study. RT-PCR assay became positive for the second time in 45 (32.8%). Anti-COVID-19 IgM and IgG antibodies were positive in 83 (60.6%) and 86 (62.8%) patients, respectively. Finally, it was determined that serological test was diagnostic in 73% of patients and the diagnostic yield of serology was significantly higher after the first week of illness (54.8% in the first week and 88% after that). Taking advantage of both serological tests and RT-PCR helps in diagnosing 83.9% of cases. Based on the present study, the serology may be useful as a complementary test and in parallel to RT-PCR assay for diagnosis of COVID-19 among admitted symptomatic cases.

ATLM (Apparent Time Lag Model) was extended to simulate the spread of infection in a mixed state of the variant virus and original wild type. It is applied to the 4th wave of infection spread in Tokyo, and (1) the 4th wave bottoms out near the end of the state of emergency, and the number of infected people increases again. (2) The rate of increase will be mainly by d strain (L452R) virus, while the increase by a strain (N501Y) virus will be suppressed. (3) It is anticipated that the infection will spread during the Olympic Games. (4) When variant viruses compete, the infection of highly infectious virus rises sharply while the infection by weakly infectious ones has converged. (5) It is effective as an infection control measure to find an infected person early and shorten the period from infection to quarantine by PCR test or antigen test as a measure other than the vaccine.

Chlamydiosis is a public health important zoonosis, and its transmission to humans occurs mainly through contact with pet birds. These animals rank the second position in most popular pets in Brazil, where the occurrence of chlamydiosis is underestimated, although studies indicate that the disease is endemic in the country. Therefore, the present work aimed to investigate the occurrence of Chlamydia psittaci in 68 cockatiel chicks (Nymphicus hollandicus) of the city of Uberlandia, Minas Gerais, Brazil. Samples of choana, cloaca, and fresh fecal samples were collected for the detection of the bacteria by PCR test. The occurrence found was null. This work concluded that it is important to raise the awareness of owners and breeders about measures to prevent and control this disease, and that orienting them about the risks of overusing antibiotics is a public health issue., A clamidiose é uma zoonose de importância na saúde pública e sua transmissão para humanos ocorre, principalmente, por meio do contato com aves de estimação. Esses animais ocupam o segundo lugar no ranking de animais de estimação no Brasil, onde a ocorrência da clamidiose é subestimada, apesar de estudos indicarem que a doença é endêmica no país. Diante disso, o presente trabalho teve como objetivo investigar a ocorrência de Chlamydia psittaci em 68 aves de estimação da espécie calopsita (Nymphicus hollandicus) da cidade de Uberlândia-MG. Foram coletadas amostras de coana, cloaca e amostras fecais frescas para a detecção da bactéria por meio do teste da PCR. A ocorrência encontrada foi nula. Este trabalho concluiu que a conscientização dos tutores e criadores quanto à adoção de medidas para a prevenção e controle dessa doença é importante, e que a orientação destes quanto aos riscos do uso indiscriminado de antimicrobianos é uma questão de saúde pública. &nbsp

The gold standard for the diagnosis of coronavirus disease 2019 (COVID-19) is a nucleic acid detection test for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which may occasionally reveal false-positive or false-negative results. Herein, we describe a case of a patient infected with human coronavirus NL63 (HCoV-NL63) who was falsely diagnosed with COVID-19 using the Ampdirect™ 2019-nCoV detection kit (Shimadzu Corporation, Japan) and SARS-CoV-2 Detection Kit (TOYOBO co., ltd.), and was admitted to a COVID-19 hospital ward. We suspected a cross-reaction between HCoV-NL63 and SARS-CoV-2; however, the reported genome sequences of HCoV-NL63 and N1/N2 primers for SARS-CoV-2 do not correspond. Thus, the PCR result was supposed to be a false positive possibly due to contamination or human error. Although the issue of a false-negative result has been the focus of much attention to prevent the spread of the disease, a false positive is fraught with problems as well. Physicians should recognize that unnecessary isolation violates human rights and a careful diagnosis is indispensable when the results of laboratory testing for COVID-19 are unclear. Generally, in cases such as a duplicate PCR test was partially positive, either N1 or N2 alone was positive, PCR testing for two or more target regions resulted in a positive only for single region, a high cycle threshold >35 was obtained, a false positive should be suspected. Especially, when these conditions coincide, we should recognize the high likelihood of a false positive.

A door-to-door survey was organised in Cuenca, Ecuador, to determine the prevalence of COVID-19 infection and adherence of the population to COVID-19 preventive measures. A total of 2457 persons participated in the study, 584 (23.7%) reported having experienced at least one flu-like symptom since the onset of the pandemic. The maximum SARS-CoV-2 seroprevalence in Cuenca was 13.2% (CI: 12–14.6%) (IgM or IgG positive). Considering PCR confirmed infections, the prevalence was 11% (CI: 10–12.4%). There was no significant difference in seroprevalence between rural and urban areas. Participants aged 35–49 years old, living with a COVID-19 positive person, at least six people in a household, physical contact with someone outside the household, a contact with a person outside the home with flu-like symptoms, using public transport, and not having enough resources for living, significantly increased the odds for SARS-CoV-2 seropositivity. Overall, there was good adherence to COVID-19 preventive measures. Having known someone who tested positive for COVID-19, having a primary or secondary level of education, and having enough resources for living, significantly increased the odds for higher adherence. In conclusion, despite good overall adherence of the population of Cuenca with COVID-19 preventive measures, our study suggests high ongoing COVID-19 transmission in Cuenca, particularly in certain parishes. Prevention should not only focus on behavioural change, but on intensified testing strategies in demographical risk groups.

PCR was applied for the discovery of adulteration of crude and processed beef meat with horse and donkey tissue. This was performed by blending (w/w) horse or donkey meat to beef meat in an extent up to 1:10000 (0.01%). The sensitivity was resolved as high as 0.01%. All used primers showed specificity in the PCR reactions utilizing layout DNAs from three animal species. PCR application on 96 beef meat and meat product samples gathered randomly from street vendors and prominent retail markets (24 of burger, 16 of minced meat, 24 of kofta, 16 of sausage, 7 of raw meat and 9 of launcheon) uncovered 6 positive for donkey tissue (3 from sausage, 2 from minced meat and 1 from kofta) and 2 positive for horse tissue (from sausage). This basic PCR strategy effectively distinguished adulteration of raw and processed beef meat samples with horse and donkey tissue. This work also highlights on the severity of the meat adulteration problem in Egypt.

Latar belakang : Kecamatan Seram Barat merupakan salah satu wilayah di bagian timur Indonesiayang endemis malariadan dikategorikan tinggi dengan indikator API diatas angka nasional. Angka kesakitan malaria per 1000 penduduk pada tiga tahun terakhir fluktuatif dimana API tahun 2014 (22,8‰), 2015(6,147‰) dan 2016 (9,03‰) dengan441kasus klinis,248kasus positif, ditemukan 23 spesies namun belumterkonfirmasi sebagai vektor malaria. Tujuan : Menganalisis secara spasial karakteritik linkungan dan dinamika kepadatan Anophele s sp. pengharunya terhadap kejadian malaria di Kecamatan Seram Barat. Metode : Jenis Penelitian ini adalah observasional analitik, desainnya cross sectional yang pelaksananya di Kecamatan Seram Barat terhadap 100 orang yang dipilih secara purposive sampling. Pengumpulan data melalui wawancara, observasi dan Penangkapan nyamuk dengan metode upan orang didalam dan di halaman rumah. Analisis data menggunakan uji chi-square . Hasil : Hasil penelitian ditemukan 41 reponden positif malaria, 5 spesies Anopheles sp. yakni An.vagus, An.teselaltus, An.kochi, An.barbirotris, An.farauti dan tidak terkonfirmasi sebagai vektor malaria, variabel yang mempengaruhi kejadian malaria adalah Suhu udara (p= 0,022, PR = 2,082), Kelembaban (p= 0,003, PR = 3,421),Kepadatan Anopheles sp. (p=0,001, PR = 2,853), Jarak Breeding places (0,000, RP= 10,054). Kesimpulanadalahtedapat 41 kasus, 5 spesies Anophele s sp. Suhu udara, kelembaban, kepadatan Anophele s sp, jarak breeding places mempengaruhi kejadian malaria, tidak ditemukan Anopheles sp sebagai vektor malaria di Seram Barat berdasakan hasil uji PCR ABSTRACT Title: Spatial Analysis of Environmental characteristics and Dynamics of Density Anopheles sp. As The Effect on Malaria Case in West Seram District, Western Area of Seram Regency, Maluku . Background : West Seram District is one of the regions in eastern Indonesia that became malaria endemic area and categorized as high with the API indicator above the national figure. The number of malaria morbidity, per 1000 of population, had been fluctuating in last three years which shown by API in 2014 (22.8 ‰), 2015 (6,147 ‰) and 2016 (9.03 ‰) with 441 clinical cases, 248 positive cases, 23 species have been found but it has not been confirmed yet as a malaria vector . T he purpose of the study is to analyze spatially the characteristics of the environment and the dynamics of the density from Anopheles sp. as the effect on the case of malaria in West Seram District. Methods: the type of this research is boservational analytic with cross sectional design. The research was held in West Seram District toward 100 people that were selected by purposive sampling.The collecting data had been done through interview, observation, and catching the mosquitoes using bait people method inside and outside the house yard. The analysis were using chi-square test. Result : The results of the study found 41 respondents positive for malaria, 5 species of Anopheles sp. namely An.vagus, An.teselaltus, An. kochi, An. barbirotris, and An. farauti. The variables affecting the case of malaria were air temperature (p = 0.022, PR = 2.082), humidity (p = 0.003, PR = 3.421), density of Anopheles sp. (p = 0.001, PR = 2,853), breeding places distances (0,000, RP = 10,054). The result of PCR test shows that there are no species containing Plamodium were found. Conclusion; The result detected 41 cases, identified 5 species of Anopheles sp. air temperature, humidity, density of Anopheles sp, distance of breeding places affecting the case of malaria. There are no Anopheles sp were found as a malaria vector in Seram Barat based on PCR test result.

The aim of our work was to develop PCR test systems for the identification and differentiation of the Piscichlamydia salmonis and Clavochlamydia salmonicola, species, that are known epitheliocystis infection agents of gill and fish skin diseases, characterized by the presence of specific ‘inclusions’ in the epithelial cells of the gills. To date, the diseases of fish associated with chlamydial infections have been detected in more than 90 species of freshwater and marine fish worldwide. For now, there is no available information on the prevalence of Piscichlamydia salmonis and Clavochlamydia salmonicola, which can cause epitheliocystis of commercially important aquaculture species in Ukraine. Identification of these pathogens is possible only using molecular genetic methods. As a result of our research, we got PCR tests for the identification and species differentiation of Piscichlamydia salmonis and Clavochlamydia salmonicola. The use of diagnostics for the identification of Piscichlamydia salmonis and Clavochlamydia salmonicola makes chlamydial infections monitoring among various fish species possible and it will increase the economic efficiency of fish farms.

Introduction Bovine tuberculosis is a chronic contagious disease caused by Mycobacterium bovis or Mycobacterium caprae. Before widespread action conducted in Poland between 1959-1975 to combat bovine tuberculosis (BTB), about 40% of all tuberculosis cases in pigs was caused by the bovine bacillus. At the present time, correctly carried out, long-term control of cattle has resulted in cases of bovine tuberculosis in pigs and humans being extremely rare and sporadic. In pigs, tuberculosis is most often caught in a slaughterhouse during slaughter. Material and methods Samples came from pigs kept on the farm. Traditional bacteriological methods on solid media (Stonebrink, LJ with pyruvate) supported by the semi-automatic, liquid indicative culture method (MGIT) and PCR test were applied in targeted studies. The GenoType Mycobacterium MTBC and CM tests (Hain Lifescience, Germany) were used to additionally confirm that isolated strains classification was used. Results Strains of mycobacteria were isolated from all examined pigs. Mycobacterium bovis was determined by real time PCR and Hain Genotype methods. Conclusions In order to effectively fight against BTB, all animals on farms should be tested, regardless of species, while the milk of suspected cows should be utilized without being used for feed. It is important to adapt the current legal regulations to the current epidemiological situation.

Leaf spot of Brassica is an important bacterial seed borne disease caused by X. campestris pv. raphani (Xcr), one of the three pathovars of Xanthomonas campestris species which makes significant economic loss throughout the world. A rapid and reliable detection method will be helpful for early diagnosis of the disease. To do that, we designed Xcr specific sequence-characterized amplified region (SCAR) markers from the Xcr specific genomic fragments, identified by aligning the whole genome sequences of several closely related bacterial pathovars and other bacterial species. Our primer set Xcr_59 produced Xcr specific amplicon of 679 bp in PCR assays. This primer pair was also capable of detecting Xcr strains directly from the artificially infected leaf samples prepared by suspending the diseased leaf area in sterile water without extracting bacterial DNA. This rapid, sensitive and reliable detection technique will be useful for adopting any firsthand plant protection measures against the disease.

The purpose of the present study was to create a real-time PCR test system allowing simultaneous detection of nontuberculous mycobacteria (NTM) and Mycobacterium tuberculosis complex (MTBC) both in culture and sputum. NTM cultures (18 strains, 18 species), MTBC cultures (16 strains, 2 species) and non-mycobacterial microorganisms from the collection of the Central Research TB Institute (CTRI) were used for the preliminary evaluation of the test system. 301 NTM cultures from patients with mycobacteriosis were used to assess the sensitivity of the developed test system. Clinical respiratory samples (sputum) from 104 patients with mycobacteriosis, 3627 patients with tuberculosis and 118 patients with other lung diseases were used for diagnostic sensitivity and specificity testing. The specificity and sensitivity of the assay for MTBC was found to be 100% both in culture and sputum samples; for NTM, the specificity was 100% in culture and sputum, the sensitivity reached 100% in culture and 73.1% in sputum samples. Positive predictive value (PPV) and negative predictive value (NPV) of the assay for culture were both 100%, for clinical material 100% and 80.8%, respectively. The limit of detection at the probability of detection 95% (LoD95%) was estimated to be 16 cfu/ml for M. tuberculosis H37RV and 1200 cfu/ml for M. avium.

Seroprevalence survey, for antibodies to SARS-CoV-2, of healthcare workers (HCW) working in three Government run hospitals in Mumbai was carried out in June 2020. Among the 801 HCWs tested, seroprevalence was 11.1%. Males (13.5% vs. 8.9% in females) and ancillary workers (18.5% vs 6.9% in doctors and nurses) were more likely to be seropositive. Sixty-two (7.74%) had been previously diagnosed with RT PCR test for SARS-CoV-2. Of these, 44 (71%) were seronegative. Upto 28 days after a positive PCR test, 90% of subjects were found to be seropositive. This reduced to less than half (38.5%) between 29 and 42 days. None of 28 infected HCWs who had the RT-PCR more than 50 days ago tested positive for antibodies. It seems likely that cellular immunity plays a larger role in defence against the illness.

Several mutant strains of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are emerging. Mismatch(es) in primer/probe binding regions would decrease the detection sensitivity of the PCR test, thereby affecting the results of clinical testing. In this study, we conducted an in silico survey on SARS-CoV-2 sequence variability within the binding regions of primer/probe published by the Japan National Institute of Infectious Diseases (NIID) and Centers for Disease Control and Prevention (CDC). In silico analysis revealed the presence of mutations in the primer/probe binding regions. We performed RT-PCR assays using synthetic RNAs containing the mutations and showed that some mutations significantly decreased the detection sensitivity of the RT-PCR assays.Our results highlight the importance of genomic monitoring of SARS-CoV-2 and evaluating the effects of mismatches on PCR testing sensitivity.

In diphtheria laboratory examinations, the PCR test can be applied to isolates and clinical specimens. This study aimed to develop a PCR assay to identify the species and toxigenicity of diphtheria-causing bacteria, including the prediction of some NTTB types. Seven reference isolates, four synthetic DNA samples, 36 stored isolates, and 487 clinical samples used for PCR optimization. The PCR results was confirmed by DNA sequence analysis. The results of the PCR examination of the 7 reference isolates and 36 stored isolates were similar to the results obtained using conventional methods as gold standard, both for diphtheria-causing and non-diphtheria-causing bacteria. The validation of the PCR results using DNA sequence analysis showed that there was no mispriming or misamplification. The multiplex PCR assay developed in this study could correctly identify the species and toxigenicity of diphtheria-causing bacteria, including the prediction of some NTTB types not yet covered by established PCR methods.

Aim To evaluate the serological response against SARS-CoV-2 in a multicenter study representative of the Spanish COVID pandemic. Methods IgG and IgM + IgA responses were measured on 1466 samples from 1236 Spanish COVID-19 patients admitted to the hospital, two commercial ELISA kits (Vircell SL, Spain) based on the detection of antibodies against the viral spike protein and nucleoprotein, were used. Results Approximately half of the patients presented antibodies (56.8% were IgM + IgA positive and 43.0% were IgG positive) as soon as 2 days after the first positive PCR result. Serological test positivity increased with time from the PCR test, and 10 days after the first PCR result, 91.5% and 88.0% of the patients presented IgM + IgA and IgG antibodies, respectively. Conclusion The high values of sensitivity attained in the present study from a relatively early period of time after hospitalization support the use of the evaluated serological assays as supplementary diagnostic tests for the clinical management of COVID-19.

Because prenatal diagnosis has ethical issues, it is important for pregnant women and their partners to understand the clinical significance of genetic tests before and after prenatal diagnosis. Therefore, genetic counseling should be performed for prenatal diagnosis of gene disorders. To date, human genome project develops a high-throughput genome analysis in addition to the conventional genetic diagnostic analysis including G-banding, FISH analysis, PCR test, etc. For example, the whole genome/exosome sequencing and the genome wide association study enable us to find the causative mutations/polymorphisms for a lot of gene disorders as a high-throughput genome analysis. In this section, the critical information for the genetic counseling of gene disorders is provided.

BackgroundAlthough the gold diagnostic method for COVID-19 is accepted as the detection of viral particles by reverse transcription polymerase chain reaction (RT-PCR), serology testing for SARS-CoV-2 is at increased demand. A primary aim for utilization of serological tests are to better quantify the number of COVID-19 cases including those RT-PCR samples were negative but showing clinical and radiological signs of COVID-19. In this study, we aimed to report the features of the patients that were diagnosed and treated as possible COVID-19 cases whose multiple nasopharyngeal swab samples were negative by RTPCR but serological IgM/IgG antibody against SARS-CoV-2 were detected by rapid antibody test.MethodWe retrospectively analyzed eighty suspected COVID-19 cases that have at least two negative consecutive COVID-19 PCR test and were subjected to serological rapid antibody test.ResultThe specific antibodies against SARS-CoV-2 were detected as positive in twenty-two patients. The mean age of patient group was 63.2 ± 13.1 years old with male /female ratio 11/11. Cough was the most common symptom with 90.9%. Most common presenting chest CT findings were bilateral ground glass opacities (77.2%) and alveolar consolidations (50.09%). The mean duration from symptom initiation to hospital admission, to hospitalization, to treatment initiation and to detection of antibody positivity were 8.6 ± 7.2, 11.2 ± 5.4, 7.9 ± 3.2 and 24 ± 17 days, respectively.ConclusionOur study demonstrated the feasibility of COVID-19 diagnosis based on rapid antibody test in the cases of patients whose RT-PCR samples were negative. We suggest that the detection of antibodies against SARS-CoV-2 with rapid antibody test should be included in the diagnostic algorithm in suspected COVID-19 patients.

Campylobacter fetus subsp . Venerealis (Cfv) is bacteria causing contagious genital diseases in cows called Bovine Genital Campylobacteriosis (BGC) or vibriosis. Isolation of Cfv is difficult, because the bacteria are fragile and need specific nutrients and oxygen (5-10%). The transport media is very important to maintain Cfv survival before culturing in laboratory. The aim of this study was to modify a new transport media as an alternative media for Cfv. Modification media capability was compared to Weybridge media, and Phosphate Buffered Saline (PBS). All transport media was contaminated by Cfv with concentrations of 10 5 , 10 4 , 10 3 , 10 2 , 10 1 (CFU/ml), and was stored for

Global differences in changes in the numbers of population-adjusted daily test-positive cases (NPDP) and deaths (NPDD) by COVID-19 were analyzed for 49 countries, including developed and developing countries. The changes as a proportion of national population were compared, adjusting by the beginning of test-positive cases increase (BPI) or deaths increase (BDI). Remarkable regional differences of more than 100-fold in NPDP and NPDD were observed. The trajectories of NPDD after BDI increased exponentially within 20 days in most countries. Machine learning analysis suggested that NPDD on 30 days after BDI was the highest in developed Western countries (1180 persons per hundred million), followed by countries in the Middle East (128), Latin America (97), and Asia (7). Furthermore, in Western countries with positive rates of the PCR test of less than 7.0%, the increase in NPDP was slowing-down two weeks after BPI, and subsequent NPDD was only 15% compared with those with higher positive rates, which suggested that the situation of testing might have affected the velocity of COVID-19 spread. The causes behind remarkable differences between regions possibly include genetic factors of inhabitants because distributions of the race and of the observed infection increasing rates were in good agreement globally.

In immunocompromised patients, BK Virus (BKV) reactivation may cause serious disease with high morbidity. Particularly for patient management after solid organ transplantation, monitoring of viral load in different clinical specimens is crucial to ensure early diagnosis and response to reactivation. In this study, we evaluated the clinical performance of a custom designed primer /probe set for detection of BKV on the cobas® 6800, a high-throughput platform, employing the open channel of the system for integration of a lab-developed test (LDT).A primer/probe set was optimized for the use on a high-throughput platform. Clinical performance was assessed in EDTA-plasma, serum and urine samples. Limit-of-detection (LOD) was determined by using a dilution series of BKV WHO standard. A CE-labeled PCR test (Altona Diagnostics) was used as a comparison to the assay.The LOD for the LDT BKV assay was 6.7 IU/mL. Inter-and intra-run variability (at 5 x LOD) was low (1.5 Ct in all specimens). All quality control panel specimens (Instand Germany n = 19) were correctly identified. Of 290 clinical samples tested, results were concordant for 280 samples. Sensitivity and specificity of the assay were 96 % and 98 % respectively. The quantitative analysis revealed a strong correlation (linear regression) between the CE-labelled comparator assay and the new BKV LDT assay with rCompared to a CE-IVD assay, the adapted LDT showed good analytical and clinical sensitivity and specificity for the detection and quantification of BKV in different clinical specimens. It represents a convenient solution to automate the LDT workflow with low hands-on time and thus facilitates high-throughput screening for BKV reactivation in immunocompromised patients.

Summary The novel Coronavirus SARS-CoV-2 caused an outbreak of pneumonia in Wuhan, Hubei province of China in January 2020. This study aims to investigate the effects of different temperature and time durations of virus inactivation on the results of PCR testing for SARS-CoV-2. Twelve patients at the Renmin Hospital of Wuhan University suspected of being infected with SARS-CoV-2 were selected on February 13, 2020 and throat swabs were taken. The swabs were stored at room temperature (20–25°C), then divided into aliquots and subjected to different temperature for different periods in order to inactivate the viruses (56°C for 30, 45, 60 min; 65, 70, 80°C for 10, 15, 20 min). Control aliquots were stored at room temperature for 60 min. Then all aliquots were tested in a real-time fluorescence PCR using primers against SARS-CoV-2. Regardless of inactivation temperature and time, 7 of 12 cases (58.3%) tested were positive for SARS-CoV-2 by PCR, and cycle threshold values were similar. These results suggest that virus inactivation parameters exert minimal influence on PCR test results. Inactivation at 65°C for 10 min may be sufficient to ensure safe, reliable testing.

A 77-year-old man was admitted to the internal medicine department for a 5-day history of progressive preauricular swelling. Two lines of antibiotic treatment failed to achieve any improvement. Fine needle aspiration cytology was conducted and smear staining with the Ziehl-Neelsen stain as well as a PCR test were positive for Mycobacterium tuberculosis. These results were confirmed with culture of the sample. A diagnosis of tuberculosis parotitis was made and anti-tuberculous drugs were initiated. LEARNING POINTS In patients not responding to usual antibiotic treatment, other rare causes of parotitis should be suspected. Fine needle aspiration cytology was an important procedure in establishing the diagnosis.

Numbers of ornamental transgenic fish are increasing, and some of them are illegally imported into Europe. The fish are modified to display different fluorescent colors under UV light. In this article, we propose real-time PCR methods to detect fish presenting green, yellow and red fluorescent coloring. The methods were tested with success and were able to detect illegally imported fish in two European countries. The article also discusses some practical information that can be useful for routine analysis. In addition, a real-time PCR test able to highlight the presence of fish DNA in general is proposed to check the amplifiability of the DNA extracted from common bony fish species of the teleost subclass. Finally, as the testing by PCR can take several days and rapid decisions must be taken with living organisms, we explored the potential of fluorescence microscopy as a screening test to determine whether animals are suspect or can be released.

Background: Sets of genital swabs are routinely taken from horses to screen for the presence of Taylorella equigenitalis, the cause of contagious equine metritis. Typically, two to four different sites are swabbed at a time and tested by culture or PCR. Objectives: This study explored the feasibility of pooling these swabs for a single PCR test per animal instead of testing each swab individually. Study design: In vitro. Methods: PCR signal strengths (Ct values) from 149 historical PCR positive genital swabs, together with historical data on the number of swabs in a set expected to be positive, were used to assess the suitability of pooling for screening horses for T. equigenitalis infection in the population at large. Twenty-four sets of four equine genital swabs were tested. The sets were prepared in the laboratory using one or more swabs positive for T. equigenitalis from naturally infected cases. Positive and negative swabs were selected to reflect a typical range of PCR Ct values expected in field cases of T. equigenitalis infection. These pools were tested by an established PCR to assess the impact and suitability of a PCR test on pooled swabs compared to individual swab testing, by comparing the Ct values. Results: Pooling one positive swab with three negative swabs produced a small drop in Ct value but all pools were still clearly positive. Main limitations: Large numbers of field positive horses are not available, but the proof of concept approach with laboratory prepared pools shows the method is applicable to field cases. Conclusions: It was concluded that pooling of swabs would confer no appreciable drop in the ability to detect a positive animal compared to individual swab testing; pooling is therefore a suitable alternative to individual swab testing with reduced costs. The Summary is available in Spanish – see Supporting Information.

Fusarium basal rot is a worldwide disease problem in onions, and causes substantial losses in onion production, both during the growing season and in the storage. To minimize the post-harvest losses, a protocol for screening of latent infections with pathogenic Fusarium oxysporum strains from harvested onions was developed. This protocol is based on a dual PCR test with primers specific for the fungal species and new SIX3 primers specific for the onion-pathogenic F. oxysporum strains. A pooled sample containing pieces from 50 harvested symptomless onions was prepared for the dual PCR using microwave disruption of the filamentous Fusarium fungi and Whatman FTA(TM) filter paper matrix technology, or as a reference protocol, by extracting DNA with a commercial kit. The two sample preparation protocols gave consistent results with the tested onion samples. Detection limit of the dual PCR protocol was 100 pg of F. oxysporum DNA, in a mixture with onion DNA, when the FTA card was applied. The new protocol reported here is simple and sensitive enough for routine testing, enabling the detection of latent infections in harvest lots even at the infection levels under 10%. Significance and Impact of the Study Fusarium basal rot causes serious problems in onion production. To minimize post-harvest losses, a simple protocol based on FTA(TM) technology and a dual PCR test with Fusarium oxysporum species-specific and pathogenicity-specific primers was developed. By testing pooled onion samples using this method, latent infections with F. oxysporum can be screened from a representative sample of the harvest. This screening method could be a useful tool to manage the post-harvest losses caused by latent infections with F. oxysporum and, with modification of the PCR protocol, with other Fusarium species pathogenic to onion.

Aim: The aim of this study was to detect Brucella spp. DNA in milk samples collected from seronegative cows using the real-time polymerase chain reaction (PCR) assay for diagnosis of brucellosis in seronegative dairy cows to prevent transmission of disease to humans and to reduce economic losses in animal production. Materials and Methods: In this study, 65 milk samples were investigated for the detection of Brucella spp. The detection of the IS711 gene in all samples was done by real-time PCR assay by comparative cycle threshold method. Results: The results show that of the 65 DNA samples tested, 2 (3.08%) were positive for Brucella infection. The mean cyclic threshold values of IS711 real-time PCR test were 37.97 and 40.48, indicating a positive reaction. Conclusion: The results of the present study indicated that the real-time PCR appears to offer several advantages over serological tests. For this reason, the real-time PCR should be validated on representative numbers of Brucella-infected and free samples before being implemented in routine diagnosis in human and animal brucellosis for controlling this disease.

Toxoplasma gondii infection can result in toxoplasmosis and potential psychological effects. Research commonly focuses on infection through contact with cat fecal matter or consumption of contaminated meat. However, T. gondii oocysts can persist in the environment for years and may be present in soils and on soil-grown produce. Rates of oocyst DNA recovery from produce were high, with 18% of vegetable samples testing positive for T. gondii via PCR test and melt curve analysis. Radishes had significantly higher oocyst counts than arugula, collard greens, kale, lettuce, and spinach. There were no significant differences in oocyst detection rates between samples taken from organic farmer's markets and conventional grocery stores. This study demonstrates that these oocysts can transfer to produce grown both conventionally and using organic techniques.

We evaluated the accuracy and ease of operation of three methods to identify tdh-positive Vibrio parahaemolyticus isolates, including the Kanagawa phenomenon test (KP test), a tdh gene PCR test, and a colloidal gold immunochromatographic assay (CGIA). A total of 221 V. parahaemolyticus isolates were collected from patients, freshly harvested seafood, and fresh seawater. Using the KP test, 92% of V. parahaemolyticus isolates from patients were identified tdh-positive, including four weak KP-positive isolates. The PCR test and CGIA also identified 92% of the isolates as tdh-positive. However, PCR and CGIA only confirmed one of the four weak KP-positive isolates. Similar results were obtained using the three methods to identify V. parahaemolyticus isolates from the other sources. Among the three methods, the KP test was the simplest to perform because it lacked any requirement for sample pretreatment, and was low cost, with no equipment requirements. Therefore, the KP test has been applied widely in many first-line quarantine laboratories. However, the sensitivity and accuracy of KP test were lower than those of the other two methods. PCR can identify the tdh rapidly, specifically, and sensitively. However, PCR requires equipment and facilities that are unavailable in first-line quarantine laboratories. The CGIA can compensate for the disadvantages of the other two methods by its higher sensitivity, accuracy, and ease of operation. Therefore, the CGIA has the highest potential to be used to identify tdh-positive V. parahaemolyticus isolates to guarantee food safety.

Several real-time PCR tests for the detection of Ralstonia solanacearum have been developed in recent years. Only the RS primer-probe system, developed by Weller et al., detects all phylotypes of R. solanacearum in one test. The Saxon State Company for Environment and Agriculture (BfUL) has been using this real-time PCR test since 2012 for routine testing of potato samples for R. solanacearum. Since the introduction of this test in the laboratory, samples were analysed which were suspected to be false positives [as they gave negative results in other standard tests of the European Union (EU) Commission Directive 2006/63/EC]. Advenella kashmirensis was identified as the cause of selected false positive samples. Inclusion of the three other known Advenella species revealed that this genus could be responsible for false positive results. Because of the rising number of these cases over the last years, two different modifications of the original test from Weller et al. were evaluated independently. It was possible to reduce false positive events, caused by Advenella species by 95% in retested potato tuber samples by using a shortened RS-primer set in the first adaption of the RS primer-probe system of Weller et al. The combination of the original RS primer-set, published by Weller et al., with the RSP-55T MGB-probe, published recently in Vreeburg et al., in a second adaption, eliminated false positive events completely.

Introduction: Imipenem-resistant Pseudomonas aeruginosa is an organism expressing metallo-β-lactamase (MBL) enzyme, and is a serious agent of hospital infection holding a serious universal therapeutic challenge. Carbapenems are potent options for the treatment of P. aeruginosa infections. The rate of MBLs expression has been variable among imipenem-resistant P. aeruginosa isolates. In the present study, we investigated the presence of MBL in the clinical isolates of P. aeruginosa. Methods: A total of 60 P. aeruginosa isolates were obtained from Kerman hospitals during 2014-2015. The antibiotics susceptibility was assessed using disk diffusion test. MBL positivity in P. aeruginosa was investigated using double disk synergy test (DDST) and polymerase chain reaction (PCR) with amplification of blaVIM-2, blaVIM-1 and blaSPM-1. Results: From 60 P. aeruginosa isolates, 28 (46.6%) were imipenem-resistant. Among these, 17 (60.7%) were identified as MBL-producing P. aeruginosa isolates using DDST. Results of PCR test demonstrated the existence of 8 (28.5%) P. aeruginosa, producing blaSPM-1. Conclusion: The frequency of blaSPM-1-producing P. aeruginosa isolates from Kerman Hospitals was relatively high. Therefore, it is recommended that the distribution of MBL-mediated resistances be managed.

The low pathogenic avian influenza becomes endemic in Egypt since its first record in 2011. It affected all the chicken sectors. There are some commercial vaccines either imported or local vaccine. This study conduct to compare the effectiveness of two H9ND vaccines, one imported and the other local, through two experiment the first in the laboratory using 150 one day old broiler chicks divided into 3 groups , the first two groups were vaccinated by the imported and local vaccine, respectively at one day old, the third group kept as negative control , the antibodies against H9N2 virus were measured by HI test at 14,21 and 28 day old , at 21 day old each group were divided to two group the 1st kept unchallenged, the 2nd challenged with H9N2 virus using the circulating local strain , in the chicken isolators ,cloacal and tracheal swaps were taken at 2,4and 6 days post challenge to detect the shedding virus using real time RT- PCR test. The second experiment were in a commercial broiler flock contain 432,000 chickens placed in 16 pens. The broiler chickens divided into 2 groups, each group took a different commercial H9ND vaccine at 1st day, the humeral immunity measured, at 14,21and 28 day old. . The results showed that the locally produced vaccine provide significant higher immune response in both lab. and field experiment ,on the other side the virus shedding is stopped early in the group vaccinated by local vaccine.

In 2014, a botulism outbreak in a flock of laying hens was investigated in France. In the flock of 5020 hens, clinical signs of botulism occurred at 46 weeks of age. A type C/D botulism outbreak was confirmed using the mouse lethality assay for detection of botulinum toxin in serum and a real-time PCR test to detect Clostridium botulinum in intestinal contents. The disease lasted one week with a mortality rate of 2.6% without recurrence. Botulism in laying hens has rarely been reported. Five monthly visits were made to the farm between December 2014 and May 2015 for a longitudinal study of the persistence of C. botulinum in the poultry house after the outbreak, and to assess egg contamination by C. botulinum. Several samples were collected on each visit: in the house (from the ventilation circuit, the egg circuit, water and feed, droppings) and the surrounding area. Thirty clean and 30 dirty eggs were also swabbed at each visit. In addition, 12 dirty and 12 clean eggs were collected to analyse eggshell and egg content. The samples were analysed using real-time PCR to detect type C/D C. botulinum. The bacterium was still detected in the house more than 5 months after the outbreak, mostly on the walls and in the egg circuit. Regarding egg contamination, the bacteria were detected only on the shell but not in the content of the eggs. Control measures should therefore be implemented throughout the egg production period to avoid dissemination of the bacteria, particularly during egg collection.

Background Feline immunodeficiency virus (FIV) infection is an important cause of disease of cats worldwide. Initial screening is commonly performed by commercially available point-of-care (POC) ELISA tests. Confirmatory testing for positive POC test results is recommended. Polymerase chain reaction (PCR) tests for FIV are commonly used additional testing methods; however, reported measures of diagnostic accuracy vary widely between PCR tests, making interpretation of results difficult. Hypothesis/Objective There is very good agreement between results of a commercially available PCR test and a POC ELISA test for FIV for specimens collected from owned and shelter-housed cats. Animals Blood samples from 168 cats from 2 adoption guarantee shelters, an FIV Sanctuary, and 64 private homes were used. Methods This was a prospective study. Whole blood samples were collected in K2-EDTA, divided, and submitted for PCR and ELISA testing. Follow-up whole blood samples were collected in lithium heparin from cats with discordant results and submitted for virus isolation (VI). Results There was very good agreement between ELISA and PCR (kappa 0.87; P < .001; 95% CI 0.79, 0.95). Of 168 cats, eleven had discordant ELISA/PCR results: 7 ELISA+/PCR- and 4 ELISA-/PCR+. Using VI as a reference standard, there were 4 false-positive PCR results, 5 false-positive ELISA results, and 1 false-negative PCR result (1 cat lost to follow-up). Conclusions and Clinical Importance While there was good agreement between the POC ELISA and PCR tests, the discordant results highlight the importance of cautious interpretation of test results and the necessity of confirmatory testing.

La leucosis bovina enzoótica (LBE) es ocasionada por un virus de la familia Retroviridae, el virus de la leucosis bovina (VLB), que afecta bovinos de cualquier edad, sexo y raza y genera importantes pérdidas económicas. En Colombia, los pocos estudios moleculares se concentran en ganado de leche; por ello, el presente trabajo se dirigió a detectar el VLB mediante una prueba molecular de PCR en animales destinados a diferentes tipos de explotación ganadera y de diferentes regiones del país, con el propósito de evaluar la relación entre la presencia del VLB en los animales, la ubicación geográfica y el tipo de explotación bovina. De un total de 230 animales, organizados por conglomerados según la región de origen, el 22.6 % se detectó con el VLB; de estos la región Centro presentó el mayor número de animales infectados (50.7 %). En cuanto al tipo de producción, el ganado de leche fue el más susceptible a ser infectado por el VLB (50.7 %). Los resultados indican que existe una significativa relación entre la presencia molecular del virus, la ubicación geográfica de los animales y el tipo de explotación bovina, datos importantes para la planeación de programas de prevención y control de la LBE por los organismos gubernamentales de salud animal.