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Objective: To identify the clinical, laboratory, and radiological markers that could predict a positive GeneXpert result in patients suspected of tuberculous meningitis. Methods: In this prospective, observational study, patients with tuberculous meningitis were systematically evaluated. Various clinical, laboratory [including cerebrospinal fluid (CSF) microscopy, culture, and GeneXpert], and neuroimaging factors were examined. All participants were administered anti-tuberculous treatment and corticosteroids. A six-month follow-up was done to evaluate the outcome. Results: Of the 116 patients studied, 54 (46.6%) tested positive for GeneXpert, while 62 (53.4%) were negative. Third cranial nerve involvement (OR 3.71, 95% CI 1.052-13.09, P =0.04) and the presence of basal exudates on neuroimaging (OR 5.22, 95% CI 2.03-13.42, P =0.001) emerged as independent predictive factors for a positive GeneXpert result. A positive CSF GeneXpert result (P =0.002) and drug resistance (P =0.004) were significantly linked to adverse outcomes. Additionally, a high score on the Medical Research Council stage III (OR 5.64, 95% CI 1.18-26.87, P =0.03) and elevated CSF cell counts (OR 1.002, 95% CI 1.00-1.001, P =0.03) were identified as independent predictors of poor prognosis. Conclusions: Third cranial nerve involvement and the presence of basal exudates were significant indicators of a positive GeneXpert result. MRC stage III disability and elevated CSF cell counts predicted poor outcomes. [ABSTRACT FROM AUTHOR]

Objective: To identify the clinical, laboratory, and radiological markers that could predict a positive GeneXpert result in patients suspected of tuberculous meningitis. Methods: In this prospective, observational study, patients with tuberculous meningitis were systematically evaluated. Various clinical, laboratory [including cerebrospinal fluid (CSF) microscopy, culture, and GeneXpert], and neuroimaging factors were examined. All participants were administered anti-tuberculous treatment and corticosteroids. A six-month follow-up was done to evaluate the outcome. Results: Of the 116 patients studied, 54 (46.6%) tested positive for GeneXpert, while 62 (53.4%) were negative. Third cranial nerve involvement (OR 3.71, 95% CI 1.052-13.09, P=0.04) and the presence of basal exudates on neuroimaging (OR 5.22, 95% CI 2.03-13.42, P=0.001) emerged as independent predictive factors for a positive GeneXpert result. A positive CSF GeneXpert result (P=0.002) and drug resistance (P=0.004) were significantly linked to adverse outcomes. Additionally, a high score on the Medical Research Council stage III (OR 5.64, 95% CI 1.18-26.87, P=0.03) and elevated CSF cell counts (OR 1.002, 95% CI 1.00-1.001, P=0.03) were identified as independent predictors of poor prognosis. Conclusions: Third cranial nerve involvement and the presence of basal exudates were significant indicators of a positive GeneXpert result. MRC stage III disability and elevated CSF cell counts predicted poor outcomes.

Sputum microscopy is the most important tool for the detection of Mycobacterium tuberculosis in peripheral laboratories. Sodium hypochlorite concentration technique prior to sputum microscopy may improve the detection of AFB bacilli over direct Ziehl Neelsen’s staining method. A prospective study was conducted at Kasturba Medical College, Manipal. Consecutive sputum samples were collected for 3 months. ZN stained smears were made directly of fresh specimens and of specimens that were processed with 3.5% sodium hypochlorite (NaOCl), from each patient. Then, ZN stained smears were observed under light microscopy. A total of 239 samples were collected. The yield of positive smears with ZN staining after NaOCl centrifugation was higher than that with microscopy after direct ZN stain. The percentage increase yield of ZN staining after NaOCl centrifugation over direct ZN staining was 3.3% (8/239). Bleach centrifugation can help in increasing the yield of positivity in sputum smear.

The preparation of myxosporeans for the description of myxospores and their preservation as type material in parasitological collections show great variations. Most frequently, formalin and ethanol are used for fixation and Giemsa solution for staining spores. In this work, authors studied the effect of 80% ethanol and 10% formalin fixation, freezing at −20 °C and staining on the size and transparency of two Myxobolus species of cyprinid fishes, M. bramae and M. bliccae spore, and recommended a new method for the deposition of type material to parasitological collections in museums. The studies have commended that fresh spores from mature plasmodia are the best material for measuring the size and studying the inner structures, the number of polar tubules in polar capsules and the morphological characters of the intercapsular appendix. The obtained quantitative data suggest that cryo- and chemical preservation do not have a notable negative effect on spores compared to fresh samples but they decrease the transparency of spores. Staining the spores with Ziehl–Neelsen has proved to be a useful method for studying the fine structure without size reduction, while Giemsa staining induced a shrinkage of spores so it seems to be not ideal for description of a new species. When treating spores of Myxobolus spp. with Lugol's solution, iodinophilous vacuoles in the sporoplasm were not recognised but visualisation of the coils of polar tubules was enhanced. As a type material for newly described species, authors suggest phototypes and spores fixed in 80% ethanol to be deposited into collections, as this preservation method is suitable for subsequent research, such as re-measurements and molecular analysis.

Tuberculosis (TB) is a multisystemic contagious disease produced by Mycobacterium tuberculosis complex bacteria (MTBC), with a prevalence of 65:100,000 inhabitants in Romania (six times higher than the European average). The diagnosis usually relies on the detection of MTBC in culture. Although this is a sensitive method of detection and remains the "gold standard", the results are obtained after several weeks. Nucleic acid amplification tests (NAATs), being a quick and sensitive method, represent progress in the diagnosis of TB. The aim of this study is to assess the assumption that NAAT using Xpert MTB/RIF is an efficient method of TB diagnosis and has the capacity to reduce false-positive results. Pathological samples from 862 patients with TB suspicion were tested using microscopic examination, molecular testing and bacterial culture. The results show that the Xpert MTB/RIF Ultra test has a sensitivity of 95% and a specificity of 96.4% compared with 54.8% sensitivity and 99.5% specificity for Ziehl–Neelsen stain microscopy, and an average of 30 days gained in the diagnosis of TB compared with bacterial culture. The implementation of molecular testing in TB laboratories leads to an important increase in early diagnostics of the disease and the prompter isolation and treatment of infected patients. [ABSTRACT FROM AUTHOR]

Sputum microscopy is the most important tool for the detection of Mycobacterium tuberculosis in peripheral laboratories. Sodium hypochlorite concentration technique prior to sputum microscopy may improve the detection of AFB bacilli over direct Ziehl Neelsen’s staining method. A prospective study was conducted at Kasturba Medical College, Manipal. Consecutive sputum samples were collected for 3 months. ZN stained smears were made directly of fresh specimens and of specimens that were processed with 3.5% sodium hypochlorite (NaOCl), from each patient. Then, ZN stained smears were observed under light microscopy. A total of 239 samples were collected. The yield of positive smears with ZN staining after NaOCl centrifugation was higher than that with microscopy after direct ZN stain. The percentage increase yield of ZN staining after NaOCl centrifugation over direct ZN staining was 3.3% (8/239). Bleach centrifugation can help in increasing the yield of positivity in sputum smear. [ABSTRACT FROM AUTHOR]

Daisuke Fukushi,1 Keigo Murakami,2 Yuji Watanabe,3 Norihiko Sugimoto,4 Hirotsugu Uehara,4 Masafumi Seki5 1Division of Gastroenterology, Tohoku Medical and Pharmaceutical University Hospital, Sendai City, Miyagi, Japan; 2Division of Pathology, Tohoku Medical and Pharmaceutical University Hospital, Sendai City, Miyagi, Japan; 3Laboratory for Clinical Microbiology, Tohoku Medical and Pharmaceutical University Hospital, Sendai City, Miyagi, Japan; 4Fuso Pharmaceutical Industry, Osaka City, Osaka, Japan; 5Division of Infectious Diseases and Infection Control, Tohoku Medical and Pharmaceutical University Hospital, Sendai City, Miyagi, JapanCorrespondence: Masafumi Seki, Division of Infectious Diseases and Infection Control, Tohoku Medical and Pharmaceutical University, 1-15-1 Fukumuro, Miyagino-ku, Sendai City, Miyagi, 983-8536, Japan, Tel +81-22-983-1221, Fax +81-22-290-8959, Email m-seki@tohoku-mpu.ac.jp; seki@hosp.tohoku-mpu.ac.jpBackground: Detection of Mycobacterium avium complex (MAC) in tissue is essential for the diagnosis of MAC infections when the Mycobacterium is not isolated from sputum. However, detection of MAC in paraffin-embedded sections has not been established.Methods: We encountered two patients with suspected MAC infections after surgery: patient 1 had a pulmonary nodule that was initially suspected to be lung cancer and was excised under video-assisted thoracoscopic surgery (VATS). Patient 2, who was under treatment with steroids and anti-IL-6 inhibitors for rheumatoid arthritis, was suspected to have disseminated ileocecal cancer with metastasis to the lung and skin. In both cases, we postoperatively detected MAC genes in paraffin-embedded tissue sections using the novel mycobacterial nucleic acid identification test, ie tissue-direct polymerase chain reaction (tdPCR)-based nucleic acid lateral flow immunoassay (NALFIA). Both patients showed granulomatous lesions with hematoxylin-eosin staining, and mycobacteria by Ziehl–Neelsen staining in tissue sections from the lung and skin, respectively, although MAC were not isolated from the sections. MAC genes were finally detected by tdPCR-NALFIA in both cases.Conclusion: Although Ziehl–Neelsen staining and culture tests are the gold standard in identifying causative mycobacteria, the rapid results of tdPCR-NALFIA performed simultaneously with sputum and/or tissue culture may make it an important auxiliary diagnostic tool for identifying mycobacterial infection, leading to improvement in the management of MAC patients.Keywords: Mycobacterium avium complex, MAC, Ziehl–Neelsen staining, rapid diagnosis

Background: Extrapulmonary tuberculosis (EPTB) is still a major health problem in developing countries. Diagnosis of EPTB is difficult due to paucity of acid-fast bacilli in clinical specimens. Objectives: To study the diagnostic role of cartridge-based nucleic acid amplification test (CBNAAT) and rifampicin resistance in EPTB on fine-needle aspiration samples. Materials and Methods: It is a prospective study comprising 79 patients of clinically suspected EPTB cases for 1 year duration. All patients underwent fine-needle aspiration cytology (FNAC) with Ziehl–Neelsen (ZN) staining followed by CBNAAT for confirmation of tuberculosis (TB) and to detect rifampicin resistance. Data have been collected and analyzed regarding sensitivity, specificity, positive predictive value, and negative predictive value of FNAC in comparison with CBNAAT. Results: Out of 79 patients, 47 (59.49%) patients were male and cervical lymph node was the most common site of aspiration (42, 52.16%). Granuloma with necrosis (38, 48.10%) followed by necrosis only (14, 17.72%) were the common findings on FNAC. Out of 79 suspected EPTB patients, CBNAAT confirmed 62 patients as having TB. Specificity of FNAC with ZN staining was 100.0%; however, sensitivity was low (43.55%) compared to CBNAAT. Only three cases were detected as rifampicin resistant. Conclusion: Compared to FNAC, CBNAAT has very good sensitivity and specificity in the diagnosis of EPTB. It has also as added advantage of detecting rifampicin resistance at the same time. CBNAAT should be used for diagnosis of EPTB wherever the facility is available.

The preparation of myxosporeans for the description of myxospores and their preservation as type material in parasitological collections show great variations. Most frequently, formalin and ethanol are used for fixation and Giemsa solution for staining spores. In this work, authors studied the effect of 80% ethanol and 10% formalin fixation, freezing at −20 °C and staining on the size and transparency of two Myxobolus species of cyprinid fishes, M. bramae and M. bliccae spore , and recommended a new method for the deposition of type material to parasitological collections in museums. The studies have commended that fresh spores from mature plasmodia are the best material for measuring the size and studying the inner structures, the number of polar tubules in polar capsules and the morphological characters of the intercapsular appendix. The obtained quantitative data suggest that cryo- and chemical preservation do not have a notable negative effect on spores compared to fresh samples but they decrease the transparency of spores. Staining the spores with Ziehl–Neelsen has proved to be a useful method for studying the fine structure without size reduction, while Giemsa staining induced a shrinkage of spores so it seems to be not ideal for description of a new species. When treating spores of Myxobolus spp. with Lugol's solution, iodinophilous vacuoles in the sporoplasm were not recognised but visualisation of the coils of polar tubules was enhanced. As a type material for newly described species, authors suggest phototypes and spores fixed in 80% ethanol to be deposited into collections, as this preservation method is suitable for subsequent research, such as re-measurements and molecular analysis. [Display omitted] • The effect of fixation, freezing at −20 C and staining on myxospores was studied. • Model materials were myxospores of Myxobolus bramae and M. bliccae. • No changes in size were observed on frozen at −20 C and 80% ethanol- or 10% formalin-fixed spores. • A shrinkage of spores was noticed when staining spores with Giemsa solution. • Phototypes and 80% ethanol-fixed spores are recommended for collections as type material. [ABSTRACT FROM AUTHOR]

Background Tuberculosis (TB) remains a significant public health challenge, particularly in developing countries, where delayed diagnosis contributes to ongoing transmission. Ziehl-Neelsen (ZN) smear microscopy, commonly used for TB diagnosis, has limitations in sensitivity, especially in cases of extrapulmonary tuberculosis (ETB). The GeneXpert Mycobacterium tuberculosis (MTB)/rifampicin (RIF) assay, a molecular diagnostic tool, offers rapid and accurate detection of MTB and RIF resistance. This study aimed to compare the diagnostic efficacy of GeneXpert MTB/RIF with ZN staining in detecting pulmonary tuberculosis (PTB) and ETB. Methods A prospective study was conducted over two years (April 2022 to April 2024) at a tertiary care teaching hospital in India. A total of 319 clinical samples from patients with suspected PTB and ETB were analyzed. Samples underwent ZN staining and GeneXpert MTB/RIF assay. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated using mycobacterial culture as the gold standard. The chi-square test was employed to compare diagnostic accuracy, with a p-value of <0.05 considered statistically significant. Results Of the 319 samples, ZN staining was positive in 18.2% of cases, while GeneXpert was positive in 21.6%. GeneXpert demonstrated a perfect sensitivity of 100% and a specificity of 98.81%, compared to ZN staining's sensitivity of 84.85% and specificity of 99.21%. GeneXpert showed superior performance in detecting TB in both pulmonary and extrapulmonary samples, with a statistically significant difference (p<0.001). Additionally, GeneXpert identified six cases of RIF resistance. Conclusion The GeneXpert MTB/RIF assay outperforms ZN staining in diagnosing TB, offering higher sensitivity and comparable specificity. Its ability to detect RIF resistance adds significant clinical value. Despite its cost, the integration of GeneXpert into routine diagnostic workflows, particularly in high TB prevalence areas, is recommended to enhance early detection and treatment, thereby reducing TB transmission., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Mahaveer Institute of Medical Sciences and Research issued approval MIMSR/IEC/2022/3/008. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Gupta et al.)

Tuberculosis remains a global disease, killing over 2 million people each year. India accounts for two-third of the global total i.e. 26%. A good, fast, and economical test to diagnose a disease is urgently needed so that effective treatment can begin as soon as possible. Various investigations can be done for TB diagnosis and these include clinical suspicion, Chest imaging, acid-fast bacilli staining, MTB culture, serological methods and Assays for amplification of nucleic acids. For developing countries like India due to the enormous number of patients and budget restrictions, the assessment of a quick and low-cost diagnostic procedure is critical. A present prospective study was undertaken to compare Ziehl-Neelsen stain with Fluorescent stain microscopy in detecting of Mycobacterium tuberculosis in TB patient’s sputum samples. Among 274 sputum samples collected from suspected TB cases, 50 (18.2%) were positive by ZN and 116 (42.3%) were positive by fluorescent microscopy. The sensitivity, specificity, Positive predictive value (PPV) & Negative predictive value (NPV) of FM microscopy was 98%, 70%, 42%, and 99% respectively. FM microscopy was superior to ZN microscopy in detecting pulmonary tuberculosis cases as it could detect 24.1% more positive cases than ZN staining. Fluorescence staining provides better sensitivity and specificity and can detect accurately more paucibacillary cases hence is of diagnostic value and can help in the early treatment of pulmonary tuberculosis.

Degefa Bekala,1 Dawit Yihdego Reda,2 Musa Mohammed Ali2 1Bulchana Health Center, Oromia, Ethiopia; 2School of Medical Laboratory, College of Medicine and Health Sciences, Hawassa University, Hawassa, EthiopiaCorrespondence: Musa Mohammed Ali Email ysnmss@yahoo.comIntroduction: Leprosy is one of the neglected tropical diseases that affect skin and peripheral nervous system often results in severe, lifelong disabilities and deformities. Even though multidrug therapy was in place for more than 30 years to treat and prevent leprosy worldwide including Ethiopia, its epidemiology is not well studied in the West Arsi zone.Objective: The aim of this study was to determine the prevalence of acid-fast bacilli (AFB) positivity rate and associated factors among leprosy suspected cases.Methods: A health facility-based cross-sectional study was conducted among 422 leprosy suspected cases from August 2020 to December 2020. To detect AFB, skin slit specimens were collected and examined using the Ziehl-Neelsen staining technique. Socio-demographic and clinical data were collected using a structured questionnaire. Data were analyzed using Statistical Package for Social Sciences (SPSS) version 24. Logistic regression was employed to determine predictors of AFB positivity rate.Results: Acid-fast bacilli were detected among 46 leprosy suspected cases which gives a prevalence of 10.9% with 95% CI (8.2‒15.6). Suspected leprosy cases with multibacillary type were 4 times more likely to be AFB positive (p=0.021) than their counterparts. Study participants who had contact with known leprosy cases were 2 times more likely to be AFB positive (p = 0.032) and those with no formal education were 2 times more likely to be AFB positive (p = 0.03). Participants who had close contact with leprosy patients for ≥ 3 years were 8 times more likely to be AFB positive (p = 0.02).Conclusion: This study revealed a high prevalence of AFB positivity rate in the era of multidrug therapy. Types of leprosy, close contact with known leprosy cases, educational status, and duration of closer contact with leprosy cases were significantly associated with AFB positivity rate.Keywords: acid-fast bacilli, Ethiopia, Mycobacterium leprae, prevalence, West Arsi zone, Ziehl-Neelsen staining

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Background: Extrapulmonary tuberculosis (EPTB) is still a major health problem in developing countries. Diagnosis of EPTB is difficult due to paucity of acid-fast bacilli in clinical specimens. Objectives: To study the diagnostic role of cartridge-based nucleic acid amplification test (CBNAAT) and rifampicin resistance in EPTB on fine-needle aspiration samples. Materials and Methods: It is a prospective study comprising 79 patients of clinically suspected EPTB cases for 1 year duration. All patients underwent fine-needle aspiration cytology (FNAC) with Ziehl–Neelsen (ZN) staining followed by CBNAAT for confirmation of tuberculosis (TB) and to detect rifampicin resistance. Data have been collected and analyzed regarding sensitivity, specificity, positive predictive value, and negative predictive value of FNAC in comparison with CBNAAT. Results: Out of 79 patients, 47 (59.49%) patients were male and cervical lymph node was the most common site of aspiration (42, 52.16%). Granuloma with necrosis (38, 48.10%) followed by necrosis only (14, 17.72%) were the common findings on FNAC. Out of 79 suspected EPTB patients, CBNAAT confirmed 62 patients as having TB. Specificity of FNAC with ZN staining was 100.0%; however, sensitivity was low (43.55%) compared to CBNAAT. Only three cases were detected as rifampicin resistant. Conclusion: Compared to FNAC, CBNAAT has very good sensitivity and specificity in the diagnosis of EPTB. It has also as added advantage of detecting rifampicin resistance at the same time. CBNAAT should be used for diagnosis of EPTB wherever the facility is available. [ABSTRACT FROM AUTHOR]

Background Tuberculosis (TB) management and prevention of further spread is crucially based on early and prompt diagnosis. Even though conventional methods are the gold standard for detection of Mycobacterium tuberculosis (MTB), they are time consuming and do not detect nontuberculous mycobacteria in most cases. The present study evaluated Anyplex MTB/NTM real-time (RT)-PCR assay (Seegene) for TB diagnosis in pulmonary and extrapulmonary TB cases in comparison with culture on Lowenstein–Jensen as a gold standard for diagnosis of MTB. Patients and methods The study included 100 patients with suspected TB: pulmonary TB represented 68 patients and extrapulmonary TB represented 32 patients. All pathological samples were stained by Ziehl–Neelsen staining (ZNS), cultured on Lowenstein–Jensen media and processed for PCR by the Anyplex MTB/NTM RT-PCR. Results Anyplex MTB/NTM RT-PCR showed a sensitivity and specificity of 100 and 65.5%, respectively, when compared with the sensitivity and specificity of ZNS, which were 87.2 and 79.3%, respectively, in the clinically suspected pulmonary TB group. However, in the clinically suspected extrapulmonary TB group, Anyplex MTB/NTM RT-PCR had sensitivity and specificity of 100 and 79.2%, respectively, when compared with those of ZNS, which were 75 and 100%, respectively. Conclusion The use of Seegene Anyplex MTB/NTM assay for detection and differentiation of pulmonary and extrapulmonary TB in our area is more convenient and accurate compared with ZNS.

Abstract In order to detect the distribution of Mycobacterium avium subsp avium (MAA) in naturally infected domestic Pekin ducks, immunohistochemistry (IHC) and Ziehl‐Neelsen (ZN) staining were used and compared. Six organs, the liver, spleen, lung, kidney, duodenum and pectoralis muscle, were collected from naturally infected Pekin ducks. Paraffin embedded tissues were examined, and the results were compared. Statistical analysis was performed using Chi‐Square test. The results showed that the detection rates by IHC were similar with ZN staining in liver, lung, spleen and pectoralis muscle, but the detection rates by IHC were much higher than ZN staining in kidney and duodenum (p = .013, p = .0044). The liver (87.5%) and lung (81.3%) had the highest detection rates. Acid‐fast bacilli (AFB) were primarily found intracellularly in six organs using ZN staining. Similarly, the MAA antigens in those selected organs were also detected in the cytoplasm with different cell types. Specifically, MAA antigen was distributed in epithelioid macrophages and necrotic centres within the liver, lung, spleen and kidney, while they were observed in macrophages of the lamina propria and duodenal glands and degenerative myocytes in the pectoralis muscle. This comparative study provides an important insight into the distribution of MAA in infected domestic ducks and indicates that the detection rate by IHC was higher than that of ZN staining.

Introduction: Hemoptysis is one of the cardinal symptoms of pulmonary tuberculosis (PTB) as per the Revised National Tuberculosis Control Programme 2016 guidelines. Hemoptysis in tuberculosis patients usually occurs in those who have a large bacillary load like that in a cavity. Direct smear done from the sputum of a PTB suspect with hemoptysis should have a higher diagnostic yield due to the greater bacillary load in such patients. Keeping this intention in mind, our study was conducted to systematically inquire into the efficacy of a cheap and easily available test like direct sputum smear examination in PTB suspect cases of hemoptysis. Materials and Methods: This was a prospective, hospital-based, analytical study, conducted on 100 PTB suspects with hemoptysis, who were attending the Outpatient and Emergency Department of National Institute of Tuberculosis and Respiratory Diseases, New Delhi, India. Patients were enrolled over a period of 8 months. All patients underwent routine clinical and radiological evaluation and sputum examination in the form of direct smear by Ziehl–Neelsen (ZN) staining and mycobacterium growth indicator tube (MGIT) culture. Results: Most of the patients (25%) belonged to the age bracket 16–25 years followed by 24% of patients who belonged to the age group of 26–35 years. Fifty-two percent of patients had the cavity in their chest X-ray findings in which mostly were the unilateral cavity (39%). Direct sputum microscopy was found to be positive in 56% of patients. In comparison to MGIT, it had a sensitivity of 79% and a positive predictive value of 92%. Conclusion: Direct smear examination in hemoptysis patients should be a first line essential investigation as it is a good, cheap, easily available, and quick test. The yield of the test increases if the patient is young, male, with a previous history of antitubercular therapy intake and cavitary lesions on the radiological findings.

Tuberculosis remains a global disease, killing over 2 million people each year. India accounts for twothird of the global total i.e. 26%. A good, fast, and economical test to diagnose a disease is urgently needed so that effective treatment can begin as soon as possible. Various investigations can be done for TB diagnosis and these include clinical suspicion, Chest imaging, acid-fast bacilli staining, MTB culture, serological methods and Assays for amplification of nucleic acids. For developing countries like India due to the enormous number of patients and budget restrictions, the assessment of a quick and low-cost diagnostic procedure is critical. A present prospective study was undertaken to compare Ziehl-Neelsen stain with Fluorescent stain microscopy in detecting of Mycobacterium tuberculosis in TB patient's sputum samples. Among 274 sputum samples collected from suspected TB cases, 50 (18.2%) were positive by ZN and 116 (42.3%) were positive by fluorescent microscopy. The sensitivity, specificity, Positive predictive value (PPV) & Negative predictive value (NPV) of FM microscopy was 98%, 70%, 42%, and 99% respectively. FM microscopy was superior to ZN microscopy in detecting pulmonary tuberculosis cases as it could detect 24.1% more positive cases than ZN staining. Fluorescence staining provides better sensitivity and specificity and can detect accurately more paucibacillary cases hence is of diagnostic value and can help in the early treatment of pulmonary tuberculosis. [ABSTRACT FROM AUTHOR]

Introduction: Tuberculosis (TB) of the breast is a rare manifestation of extra-pulmonary localization of the disease. The incidences vary from a minimum in developed countries to a high in developing countries like India and Africa. Materials and Method: This study was a hospital-based observational study carried out in 75 cases of benign breast disease which were studied with special reference to Tuberculosis of the breast. All patients who were clinically diagnosed as a benign breast lump, above the age of 14-years and among them who were willing to undergo investigations and treatment were included. Ziehl-Neelsen staining was performed in all fine needle aspirates and finally, treatment modality used. Results: The present study involved 67 cases of benign breast lumps and 8 cases of recurrent breast abscesses. Further, it depicts predominant benign breast lumps which were fibroadenoma in 65.7% fibroadenosis in 23.80%, TB breast in 4.50%, ductal papilloma in 2.90% and phyllodes tumour of the breast in 1.5% of cases. Conclusion: For the TB of the breast in India, various treatment modalities are available. With timely diagnosis, the morbidity can be minimised with less amount of suffering. [ABSTRACT FROM AUTHOR]

Introduction: Tuberculosis (TB), caused by the bacterium Mycobacterium tuberculosis, remains one of the major health problems in India. Early detection of TB and Rifampicin (RIF) resistance are essential for effective disease management. The recent introduction of Cartridge-Based Nucleic Acid Amplification Test (CBNAAT) also known as Gene Xpert MTB/ RIF assay has significantly transformed the diagnostics of TB. Aim: To evaluate the role of CBNAAT and smear microscopy by Ziehl-Neelsen (ZN) staining in the diagnosis of Pulmonary Tuberculosis (PTB). Materials and Methods: Sputum samples from 337 patients having symptoms suggestive of PTB were included in this study. Sputum samples for ZN staining and CBNAAT were processed. RIF resistance was detected by CBNAAT. Results: Out of 337 samples, 36 (10.68%) sputum samples were positive by smear microscopy. Samples from 107 cases with high clinico-radiological presumption were subjected to CBNAAT examination, out of which 41 (38.31%) were confirmed positive microbiologically. The mean age was 45±18.30 years, 24% were females and 76% were males. Overall sensitivity of CBNAAT was 38.31%. Sensitivity of CBNAAT was 100% for sputum positive cases and sensitivity was 15.38% for sputum negative cases. Overall RIF resistance was detected in two (1.86%) cases in present study. Conclusion: CBNAAT helps in early detection to diagnose PTB. It can be used for screening MDR-TB for starting anti-tubercular treatment early.

Setting Tuberculosis (TB) still remains one of the major health problems facing humans. India accounts for almost a quarter of TB cases worldwide. The scenario is worsening owing to multidrug resistant-tuberculosis (MDR-TB). Assessment of local prevalence rates and detection of MDR-TB are important to rationalize therapy and prevent spread of resistant strains in community. Objective The study was undertaken to understand drug sensitivity patterns of tubercle bacillus and assess resistance trends in Meghalaya. Designs Specimens were screened for acid-fast bacilli, decontaminated by N-acetyl cysteine–sodium hydroxide method, and subsequently inoculated onto Lowenstein–Jensen media. Characteristic growth was biochemically identified as TB bacillus. Drug sensitivity assessment to first-line anti-TB drugs was performed by proportion method and sensitivity patterns noted. Results Among 103 specimens received, 23 showed acid-fast bacilli. Male to female ratio was 3 : 2. Fourteen (13.6%) pure isolates of Mycobacterium spp. were obtained. Biochemically 10 isolates were confirmed as M. tuberculosis. Drug sensitivity profile revealed highest mono resistance to isoniazid and streptomycin. Two (20%) isolates were MDR. Conclusion The study gives a brief overview of the menace of TB in Meghalaya. The study results provide valuable information about presence of primary MDR-TB and provide basis for future larger field surveys.

Introduction: Hemoptysis is one of the cardinal symptoms of pulmonary tuberculosis (PTB) as per the Revised National Tuberculosis Control Programme 2016 guidelines. Hemoptysis in tuberculosis patients usually occurs in those who have a large bacillary load like that in a cavity. Direct smear done from the sputum of a PTB suspect with hemoptysis should have a higher diagnostic yield due to the greater bacillary load in such patients. Keeping this intention in mind, our study was conducted to systematically inquire into the efficacy of a cheap and easily available test like direct sputum smear examination in PTB suspect cases of hemoptysis. Materials and Methods: This was a prospective, hospital-based, analytical study, conducted on 100 PTB suspects with hemoptysis, who were attending the Outpatient and Emergency Department of National Institute of Tuberculosis and Respiratory Diseases, New Delhi, India. Patients were enrolled over a period of 8 months. All patients underwent routine clinical and radiological evaluation and sputum examination in the form of direct smear by Ziehl–Neelsen (ZN) staining and mycobacterium growth indicator tube (MGIT) culture. Results: Most of the patients (25%) belonged to the age bracket 16–25 years followed by 24% of patients who belonged to the age group of 26–35 years. Fifty-two percent of patients had the cavity in their chest X-ray findings in which mostly were the unilateral cavity (39%). Direct sputum microscopy was found to be positive in 56% of patients. In comparison to MGIT, it had a sensitivity of 79% and a positive predictive value of 92%. Conclusion: Direct smear examination in hemoptysis patients should be a first line essential investigation as it is a good, cheap, easily available, and quick test. The yield of the test increases if the patient is young, male, with a previous history of antitubercular therapy intake and cavitary lesions on the radiological findings. [ABSTRACT FROM AUTHOR]

In order to detect the distribution of Mycobacterium avium subsp avium (MAA) in naturally infected domestic Pekin ducks, immunohistochemistry (IHC) and Ziehl‐Neelsen (ZN) staining were used and compared. Six organs, the liver, spleen, lung, kidney, duodenum and pectoralis muscle, were collected from naturally infected Pekin ducks. Paraffin embedded tissues were examined, and the results were compared. Statistical analysis was performed using Chi‐Square test. The results showed that the detection rates by IHC were similar with ZN staining in liver, lung, spleen and pectoralis muscle, but the detection rates by IHC were much higher than ZN staining in kidney and duodenum (p =.013, p =.0044). The liver (87.5%) and lung (81.3%) had the highest detection rates. Acid‐fast bacilli (AFB) were primarily found intracellularly in six organs using ZN staining. Similarly, the MAA antigens in those selected organs were also detected in the cytoplasm with different cell types. Specifically, MAA antigen was distributed in epithelioid macrophages and necrotic centres within the liver, lung, spleen and kidney, while they were observed in macrophages of the lamina propria and duodenal glands and degenerative myocytes in the pectoralis muscle. This comparative study provides an important insight into the distribution of MAA in infected domestic ducks and indicates that the detection rate by IHC was higher than that of ZN staining. [ABSTRACT FROM AUTHOR]

Objective: To compare the validity of FNAC & ZN staining in diagnosing tuberculous lymphadenitis taking mycobacterial culture as gold standard. Study Design: Cross sectional study. Place and Duration of Study: Pathology department, Fauji Foundation Hospital Peshawar, from Oct 2019 to Sep 2020. Methodology: This study includes 100 patients with enlarged cervical lymph nodes who presented as outdoor patients. After routine baseline & radiological investigations, FNAC & ZN staining was carried out, followed by mycobacterial culture. Results: FNAC of tuberculous patients was positive in 56 (56%) patients while 24 (24%) patients revealed posi-tive mycobacterial culture. Inpatients with positive culture, 77 (77%) were FNAC positive and 23 (23%) were negative on cytology. Conclusion: Fine needle aspiration is more sensitive & specific test than Ziehl-Neelsen staining. [ABSTRACT FROM AUTHOR]

Introduction: In developing countries, diagnosis of pulmonary tuberculosis is primarily based on microscopy owing to its simplicity, less cost and rapidity. Various ways to improve the sensitivity with multiple samples and concentration of sputum have been used. Simple methods which can be used in microscopic centers are desirable. Aim: This study was planned to evaluate the sensitivity of sputum smear examination by Pot’s technique and Sodium hypochlorite concentration technique over conventional Revised National Tuberculosis Program (RNTCP) method. Materials and Methods: A cross-sectional analytical study was carried out in all non-repetitive sputum smear negative samples obtained in Mycobacteriology laboratory after obtaining the Institutional Ethics Committee clearance with waiver of consent during a time period from 1st July to 31st August 2018. Two sputum samples (one morning and one spot) received from 47 outdoor and indoor patients in Mycobacteriology department of tertiary care hospital were included in study. All samples were first processed by routine Ziehl-Neelsen (ZN) staining. Sputum smear negative were subjected to Pot’s technique and Sodium hypochlorite concentration technique to evaluate the efficacy of these concentration techniques. All data was maintained in MS Excel and analysed using test of proportion and test of significance. Results: A total of 94 samples received from 47 patients were analysed. 04/94 (4.25%) samples were positive for AFB by routine RNTCP staining. With use of Sodium hypochlorite concentration method and Pots method, 05/94 (5.31%) and 06/94 (6.38%) samples were positive indicating a rise of 1.06% and 2.1% respectively over conventional. Additional two cases were detected compared to RNTCP, amounting to 100% rise in case detection which otherwise would have been missed. Conclusion: Concentration methods are easy to perform, safe for handlers without requirement of additional human resource. Good smear quality with less debris in microscopic field aids in improving detection rate over conventional methods.

OBJECTIVE: The objective of the study was to evaluate the utility of fluorescent staining (FS) technique with direct sputum for the diagnosis of pulmonary tuberculosis (TB). MATERIALS AND METHODS: Three sputum specimens (spot, second spot, and morning) were collected from the study participants. One smear was prepared from the direct sample and one using the N-acetyl-L-cysteine-sodium hydroxide concentration method. The direct smears were stained using FS, and Ziehl–Neelsen (ZN) staining was used to stain smears prepared after concentration method. Smear results were pooled and compared in two categories: standard spot-morning and same-day smears. The Chi-square test was used to evaluate the statistical difference in smear-positive cases. RESULTS: Among 2922 participants included in the study, smear positivity was, respectively, 10.5% and 10.2% for FS and ZN smears using spot-morning approach. Using the same-day approach, the smear positivity was, respectively, 10.4% and 9.9% for FS and ZN smears; statistically, the difference was not statistically significant (P > 0.05) between the staining techniques in both the approaches. CONCLUSION: More smear positivity is the advantage of using FS technique. Given this advantage, National TB Control Programs of high-TB-burden countries should consider installing light-emitting diode FS technique in peripheral settings also. [ABSTRACT FROM AUTHOR]

From among intestinal parasites, coccidian intestinal parasites such as Cystoisospora belli (previously known as Isospora belli), and Cyclospora cayetanensis are well-known as opportunist parasites, particularly in patients with cancer. This study assessed the prevalence of C. belli and C. cayetanensis in patients with cancer in Lorestan Province, Southwest of Iran. This cross-sectional descriptive study was conducted on 87 patients with colorectal cancers, referred to the general hospitals of Lorestan from October 2017 to August 2018. A fresh stool specimen was collected from each subject in a sterile labeled container. The collected stool samples were concentrated through sucrose flotation method and then prepared for Ziehl–Neelsen staining for microscopic examination. Demographic and possible risk factors such as age, sex, education, residence, and unwashed vegetable/fruit consumption were collected by an applied questionnaire. Out of the 87 patients with colorectal cancer, eight (9.2%) were found positive for C. belli and C. cayetanensis infections, with five (5.74%) and three (3.44%) patients positive for C. belli and C. cayetanensis infections, respectively. Results also showed that sex and unwashed vegetable/fruit consumption were significantly associated with the prevalence of these parasites (p < 0.01). The findings revealed the considerable prevalence of C. belli and C. cayetanensis in patients with colorectal cancers. It is therefore essential for gastrointestinal specialists to pay special attention to the prevalence of coccidian parasites in patients with colorectal cancer. [ABSTRACT FROM AUTHOR]

Introduction: In developing countries, diagnosis of pulmonary tuberculosis is primarily based on microscopy owing to its simplicity, less cost and rapidity. Various ways to improve the sensitivity with multiple samples and concentration of sputum have been used. Simple methods which can be used in microscopic centers are desirable. Aim: This study was planned to evaluate the sensitivity of sputum smear examination by Pot's technique and Sodium hypochlorite concentration technique over conventional Revised National Tuberculosis Program (RNTCP) method. Materials and Methods: A cross-sectional analytical study was carried out in all non-repetitive sputum smear negative samples obtained in Mycobacteriology laboratory after obtaining the Institutional Ethics Committee clearance with waiver of consent during a time period from 1st July to 31st August 2018. Two sputum samples (one morning and one spot) received from 47 outdoor and indoor patients in Mycobacteriology department of tertiary care hospital were included in study. All samples were first processed by routine Ziehl-Neelsen (ZN) staining. Sputum smear negative were subjected to Pot's technique and Sodium hypochlorite concentration technique to evaluate the efficacy of these concentration techniques. All data was maintained in MS Excel and analysed using test of proportion and test of significance. Results: A total of 94 samples received from 47 patients were analysed. 04/94 (4.25%) samples were positive for AFB by routine RNTCP staining. With use of Sodium hypochlorite concentration method and Pots method, 05/94 (5.31%) and 06/94 (6.38%) samples were positive indicating a rise of 1.06% and 2.1% respectively over conventional. Additional two cases were detected compared to RNTCP, amounting to 100% rise in case detection which otherwise would have been missed. Conclusion: Concentration methods are easy to perform, safe for handlers without requirement of additional human resource. Good smear quality with less debris in microscopic field aids in improving detection rate over conventional methods. [ABSTRACT FROM AUTHOR]

Background: A fast and accurate diagnosis is necessary to control and eliminate tuberculosis (TB). There have been various studies evaluating the efficacy of polymerase chain reaction (PCR) testing in clinical practice. Most of these studies have found PCR as a useful tool to diagnose TB and more so for the extra-pulmonary cases. Aims and Objectives: The aim of the study was to evaluate the results of TB by real-time PCR versus the conventional methods of diagnosis in a tertiary care center in Punjab. Materials and Methods: This study was done in a tertiary care center of Punjab to look for the results of clinical samples tested for TB using PCR, Ziehl–Neelsen staining and culture on Lowenstein–Jensen medium. 16S rRNA gene was used for Mycobacterium tuberculosis detection in PCR. This study included all samples tested for TB from July 2015 to October 2015. Results: A total of 214 samples were tested for TB using smear microscopy, culture and PCR (including 194 [90.6%] extra-pulmonary and 20 [9.3%] pulmonary samples). These included 76 cerebrospinal fluid, 30 pleural fluid, 42 tissues, 17 ascitic fluid, 13 urine, 15 bronchoalveolar lavage (BAL), 16 pus, and 5 sputum samples. Smear microscopy was positive in a total of 4 samples out of 214 (1.8%), which included 2 sputum, 1 tissue, and 1 ascitic fluid samples, while culture was positive for 6 samples (2.8%) which included 2 sputum, 1 pus, 1 BAL, 1 tissue, and 1 ascitic fluid sample. The TB PCR results were positive in a total of 71 (33.1%) samples out of 214. In addition, three samples also tested positive for Mycobacteria other than TB. Conclusion: PCR is a very rapid and accurate diagnostic tool for early detection of TB in particularly for extrapulmonary TB.

Background: Sputum smear microscopy is the main tool for the diagnosis of pulmonary tuberculosis (TB), especially in low- and middle-income countries (LMICs). Limited sensitivity of smear microscopy and patient dropouts (PDs) are the important obstacles of national TB control programs. Objectives: (1) To assess the diagnostic utility of the same day (SS2) approach (2) To compare the smear results of the spot morning (SM) and the SS2 approaches. Materials and Methods: The study was conducted in the Department of Microbiology, GSL Medical College, Rajahmundry, Andhra Pradesh, India from January 2011 to February 2015. Three sputum samples were collected [spot (S), second spot (S2) 1 h after S, and morning sample (M)] from the volunteers. The sputum smears were stained by Ziehl-Neelsen (ZN), modified ZN (MZN), and fluorescent staining (FS) techniques and the results were pooled and compared under SM and SS2 approaches. Results: Of the 3,186 study participants, sputum smear positivity (SSP) for SM approach was 9.6% and 10.8% and for SS2 approach, it was 9.4% and 10.6%, respectively, with ZN and FS and the results were statistically insignificant (Mann-Whitney U test, P > 0.05). Conclusion: Technically SSP was similar for both the approaches and no improvement was observed with the SS2 approach. Hence, there is an urgent need to improve SSP.

Granulomatous prostatitis is a rare inflammatory condition of prostate. The diagnosis of this entity continues to remain a clinical surprise for the treating urologist. They represent a clinical diagnostic dilemma as some of the cases mimic malignancy. The radiological findings and serum prostate specific antigen levels are non- specific in their diagnosis. The definitive diagnosis can be established only by histopathological examination. It is essential to typify the granulomatous prostatitis histologically, so as to guide the management. The presence of Langhans type and foreign body type giant cells with the presence of necrosis does not confirm the diagnosis of tubercular prostatitis, confirmation in fact requires Ziehl-Neelsen staining and tubercular polymerase chain reaction. Nonspecific granulomatous prostatitis represents the majority of the cases of granulomatous prostatitis.

The conventional methods for diagnosis of tubercular lymphadenitis (TBLN) such as - fine needle aspiration cytology, Ziehl-Neelsen staining and culture have limitations of low sensitivity and/or specificity. So, it becomes essential to develop a rapid, sensitive, and specific method for an early diagnosis of TBLN. Therefore, the present study was conducted to evaluate nested multiplex polymerase chain reaction (nMPCR) targeting MTP40 and IS6110 gene sequences of Mycobacterium tuberculosis and Mycobacterium tuberculosis complex, respectively in 48 successive patients of TBLN and 20 random patients with non-tubercular lymph node lesions. Out of the 48 cases of TBLN, 14 (29.2%) were found to be positive by Ziehl-Neelsen staining, 15 (31.2%) were positive by culture and 43 (89.6%) cases were positive after first round of PCR while 48 (100%) cases were positive by nMPCR assay. The sensitivity and specificity of nMPCR was found to be 100% for the diagnosis of TBLN. The results thus obtained indicate that nMPCR assay is a highly sensitive and specific tool for the diagnosis of TBLN. [ABSTRACT FROM AUTHOR]

Background and Methods: Paratuberculosis is a chronic disease of ruminant, caused by Mycobacterium avium subsp. paratuberculosis (Map), clinically infected animals produce high level of antibodies in blood and shed detectable amount of Map organisms in feces. Several serological and molecular tests are utilized for detection of antibodies or DNA of the organism in clinical samples. Present study indicates the status of paratuberculosis in two distinct cattle farms with different organizational set-ups viz. organized and unorganized. We used agar gel immunodiffusion (AGID) assay for the detection of antibodies in blood. Ziehl-Neelsen (ZN) staining of fecal smears was done to observe acid-fast bacilli and Nested PCR targeted to IS900 and f57 sequences, was performed to confirm the pathogen.Results: Sera samples of cattle, from organized farm, did not show any visible precipitating band with AGID assay. However, fecal smears of few cattle (3.57%) were positive for acid-fast bacilli. When confirmed with nested PCR, only one fecal sample (0.71%) was found positive for Map. In case of unorganized farm, a large number of cattle (38.75%) showed precipitating antibodies with AGID assay and the percentage of fecal smears that showed acid-fast bacilli was 26.62%. Nevertheless, fecal samples containing Map DNA was confirmed in 14.37% of fecal sample by nested PCR.Conclusions: An organized farm, with better hygiene and management practices, showed lesser occurrence of paratuberculosis in cattle in comparison to unorganized farm. Not all AGID assays positive cattle might be an efficient shedder of Map and mare detection of acid-fast bacilli in fecal smears did not always indicate the presence of Map organism. Cattle infected with JD were mostly in the age group of six years and above.

Background : Tuberculosis is the second leading cause of death worldwide, killing nearly two million people each year. Sputum decontamination with N-acetyl-L-cysteine-sodium hydroxide (NALC-NaOH) is expected to improve detection of Mycobacterium tuberculosis (M. tb) by culture better than that with modified Petroff′s; which is widely used in laboratories. In this study, sputum samples collected from suspected cases of pulmonary tuberculosis (TB) were cultured directly on Lowenstein-Jensen (LJ) medium and after decontamination by both the methods and the results of smear and culture positivity were evaluated to assess whether the NALC-NaOH treatment method improves smear and culture. Materials and Methods : For each decontamination method, 30 samples were obtained from suspected cases of Pulmonary TB, from Pad. Dr. D.Y. Patil Medical College and Hospital. Two sputum samples from each patient were collected on day 1 and 2. These samples then underwent decontamination process by performing the 4% NaOH, NALC-2% NaOH treatment methods and direct inoculation. After each process a smear was made and culture was done on LJ medium. Results: The modified Petroff′s and NALC-NaOH treatment methods did not significantly affect the AFB smear positivity of the sputum samples (66% and 72.3%, respectively). (However, the culture positivity for M. tb on LJ medium was significantly different by the three processes. With NALC-NaOH and modified Petroff′s it was 63% and 46%, respectively, while with direct culture it was 23%. Conclusion: NALC-NaOH treatment is better than modified Petroff′s treatment for the detection of M. tb by culture. However, AFB microscopy does not seem to be significantly different by either process.

Abstract Background and objectives: Direct smear microscopy, because of its simplicity, rapidity, low cost and relatively sensitive is a suitable method to detecting pulmonary tuberculosis. This experiment was aimed at determining the best method for detecting Mycobacterium tuberculosis among three kinds of staining methods: Fluorochrome, Ziehl Neelsen and Tan Thiam Hok . Material and Methods: The sputum specimens (N=714) were obtained from people with suspected pulmonary tuberculosis and identified by three staining procedures. We determined the sensitivity, specificity, and the positive and negative predictive value of them and compared results by growth on Lowenstein-Jensen medium as gold standard. Results: Ninety-three (13%) of 714 sputum specimens were positive in culture method. The sensitivity of Tan Thiam Hok, Ziehl Neelsen and Fluorochrome are 89.2%, 91.3% and 95.6%, respectively, while their specificity and positive predictive value were 100%. Their negative predictive values were 98.3%, 98.7 %, 99.3 %, respectively. Conclusion: We conclude that Ziehl Neelsen can still be a reliable procedure for detecting AFB in sputum specimens because it has the appropriate sensitivity and specificity in comparison with another method of staining. Key words: pulmonary tuberculosis, Tan Thiam Hok staining, Ziehl-neelsen staining , Flurochrome staining

Ziehl-Neelsen (Z-N) staining of cerebrospinal fluid (CSF) for acid-fast bacilli (AFB) is the cornerstone of the laboratory diagnosis of tuberculous meningitis (TBM). However, the sensitivity of conventional Z-N staining for the detection of AFB in CSF specimens is suboptimal. The present study aimed to compare the practicality of modified Z-N staining with light microscopy and fluorescence microscopy in the same smear without auramine O. A total of 155 patients with 223 CSF specimens were enrolled and grouped according to the uniform case definition. The smears of each CSF specimen were subjected to modified Z-N staining and then observed using a light microscope under transmitted light and under fluorescence with a green-excitation wavelength in the same microscopic field. The results for different groups, inspection times, and prior to and following treatment were compared. Results indicated that the fuchsin-stained AFB were visible as bright orange-red fluorescing rods under fluorescence, or as red, lightly curved rods under transmitted light. The sensitivity of fluorescence microscopy was 96.2% while that of light microscopy was 84.6%. The positive rate of fluorescence microscopy was 79.2% prior to treatment compared with 61.7% post-treatment. In the same microscopic field, a greater number of AFB were observed using fluorescence compared with transmitted light, and AFB that were not visible under transmitted light were clearly observed under fluorescence. Furthermore, transmitted light and fluorescence could be interchanged directly when equivocal smears were encountered. The combination of modified Z-N staining and fluorescence microscopy without auramine O is sensitive and convenient for the diagnosis of TBM. [ABSTRACT FROM AUTHOR]

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Objective/background: Lack of rapid and accurate diagnostic testing is a critical obstacle to global tuberculosis (TB) control. Sensitivity of sputum smear microscopy (SSM) is not optimal; however, it remains the most prevalent tool for TB confirmation in poor countries. As a part of passive case finding of TB detection, this study was conducted to determine the clinical performance of PURE TB-LAMP assay using liquid culture medium as the gold standard. Methods: Centre Antituberculeux de Yopougon is one of the 17 intermediate Tuberculosis centers in Côte d’Ivoire. A standardized questionnaire was submitted to patients with signs and symptoms consistent with tuberculosis by a trained caregiver. After obtaining signed consent forms, sputum samples were collected according to National TB Control Programme guidelines (spot-morning). SSM after Ziehl–Neelsen staining and TB-LAMP assay were blindly performed on the first sample. Samples transported to Institut Pasteur de Côte d'Ivoire were decontaminated according to the N-acetyl-L-Cystein method. In Mycobacteria Growth Indicator Tube (MGIT), 500 mL of pellets were inoculated and incubated in the MGIT 960 system. MPT64 antigen was detected in positive cultures. Results: Of the 500 patients enrolled, 469 (232men and 239 women) patients were included. The mean ages of men and women were 36.9 (15–86) and 37.3 (15–37.3) years, respectively. There were 56 (12.2%) HIV-infected patients, including 14 women. Clinical isolates of M. tuberculosis complex were detected for 157 (33.5%) patients. Compared with culturing, the overall sensitivity and specificity of SSM were 86% (95% confidence interval [CI] = 81–91) and 96% (95% CI = 94–98), respectively. The overall sensitivity and specificity for TB-LAMP was 92% (95% CI = 0.88–0.96) and 94% (95% CI = 0.91–0.97), respectively. Positive likelihood ratios for TB-LAMP and SSM were 15.3 and 21.5, respectively, and negative likelihood ratios for TB-LAMP and SSM were 0.09 and 0.15, respectively. Among the 469 patients, active tuberculosis was detected using TB-LAMP assay and SSM in 162 (34.5%) and 147 (31.3%) patients, respectively. Conclusion: For accurate diagnostic of pulmonary TB, TB-LAMP could be used as a tool of the first intention.