Sputum Smear Microscopy Research Articles

População em situação de rua: perfil dos casos de coinfecção tuberculose e HIV

OBJETIVO: analisar o perfil da População em Situação de Rua (PSR) com a coinfecção tuberculose (TB)/HIV em Porto Velho-RO, entre 2015 a 2018. MATERIAIS E MÉTODOS: estudo descritivo, transversal com abordagem quantitativa realizado a partir do levantamento das variáveis sociodemográficas, clínica e do tratamento dos casos no Sistema de Informação Nacional de Agravos de Notificação, e posteriormente análise por distribuição de frequência, após atender aos preceitos éticos. RESULTADOS: foram notificados sete casos de TB/HIV entre as PSR. A maioria era do sexo feminino, entre 31 a 38 anos, parda, baixa escolaridade, apresentavam doenças e agravos associados, não eram população especial, notificadas em 2016, na referência terciária, era caso novo ou reingresso após abandono, forma clínica pulmonar, radiografia de tórax suspeito, baciloscopia de escarro negativa, teste molecular rápido para a TB e cultura não realizadas, teste de sensibilidade e controle mensal em branco, até três contatos identificados e nenhum examinado, não realizaram Tratamento Diretamente Observado, tratamento na referência terciária, de 61 a 120 dias e mais do que 181 dias, e elevado percentual de abandono. CONCLUSÃO: torna-se essencial rever as estratégias de acesso aos serviços e ações ofertadas para atender as demandas de saúde da PSR, voltadas para ambos os agravos.

Determinants of coinfection tuberculosis and HIV in prisons in Brazil.

Prisons context has the potential for the spread of infectious diseases, like HIV and tuberculosis, which prevalence is higher in the people deprived of liberty compared to the general population. to analyze which are the determinants of coinfection tuberculosis and HIV in prisons. Case-control study conducted in the state of São Paulo, Brazil. New cases of tuberculosis in the population deprived of liberty in the period between 2015 and 2017 were considered. Data were obtained through the notification and monitoring system for tuberculosis cases in the state of São Paulo and included sociodemographic and clinical variables and diagnosis and treatment information. The data were analyzed through frequency distribution and bivariate analysis, testing the association of the dependent variable (tuberculosis/HIV coinfection vs. tuberculosis/HIV non-coinfection) with independent variables (sociodemographic, clinical and diagnostics variables) by calculating the odds ratio and p-value. Among the determinants of tuberculosis/HIV coinfection in prisons, we identified: age between 26-35, 36-55 and 56-84 years, notification in hospitals, negative sputum smear microscopy and culture, X-ray suggestive of another pathology, extrapulmonary and mixed clinical form, and alcoholism. A high percentage of death was also identified among coinfected people. identifying the determinants of the tuberculosis/HIV coinfected individual can assist in the development and implementation of guidelines aimed at controlling both infections in the prison environment.

Polyester functionalization for mycobacterial capture

AbstractTuberculosis (TB) is one of the deadliest diseases known to man and ranks among the top ten global causes of fatalities. Children often develop a paucibacillary form of this disease which makes diagnosis arduous even with the aid of sophisticated techniques. In these cases, the concentration of bacteria in their sputum falls below the lowest limit of detection for sputum smear microscopy, the technique predominantly used in developing countries. In this study we aim to test the hypothesis that modifying a polymer, often used in buccal swabs, could assist in creating a capturing and concentrating oral swab for Mycobacterium tuberculosis (Mtb), the pathogenic bacterial species responsible for TB. Such a device will assist in meeting the detection limit and allow for rapid detection of the disease. The polymer used was a micro fibrous form of poly(ethylene terephthalate) (PET) which was surface functionalized with Concanavalin A (Con A), a lectin based adhesin with an affinity for the mannose groups on the Mtb cell wall. The functionalization was mediated with glutaraldehyde as bioconjugate molecule and found to produce a surface uniformly covered with Con A. Affinity studies between the modified fibers and the Mycobacterium bovis bacillus Calmette‐Guérin (BCG), as Mtb mimic, were conducted to evaluate the capturing abilities of the substrate. The results indicate that the fibers avidly captured BCG and that the fibrous matrix aided in its function. Dilution studies showed successful capturing of the bacterial species at concentrations characteristic of paucibacillary cases. In this study a commercial polymeric material was successfully surface modified with a biological entity to create a substrate to which Mtb could adhere and accordingly be captured.

Rapid Testing for the Diagnosis of Pulmonary Tuberculosis and Rifampicin Resistance: A Review of Cost-Effectiveness


 The evidence regarding the cost-effectiveness of the Xpert Mycobacterium tuberculosis complex and resistance to rifampicin (Xpert MTB/RIF) test compared with smear microscopy in diagnosing tuberculosis is summarized in this report.
 Results from the included 6 studies showed that Xpert MTB/RIF testing is a cost-effective option compared with sputum smear microscopy. However, the generalizability of the results to the Canadian setting are unclear because of the clinical data source populations, willingness-to-pay thresholds, and assumptions used in the analyses.
 There is a lack of evidence regarding the cost-effectiveness of Xpert MTB/RIF testing compared with mycobacterial cultures or culture-based susceptibility testing.

Treatment success rates among tuberculosis patients notified from the private sector in West Bengal, India

In India, an estimated 50% of tuberculosis (TB) patients are diagnosed and managed by private providers (PPs). However, there is limited information on treatment outcomes among those treated by PPs largely because of a lack of systems for patient follow-up and documentation of the outcomes. Tuberculosis Health Action Learning Initiative (THALI) project implemented in six districts of West Bengal, supported PPs in managing TB patients and systematically documented the TB treatment outcomes of such patients. This provided us an opportunity to describe the treatment outcomes and assess factors associated with ‘unsuccessful outcomes’ among TB patients notified by PPs from January to April 2018. This was a retrospective cohort study using routinely collected data. During the study reference period, 2347 patients were notified from 389 PPs. The patient’s mean (SD) age was 39.9 (17.2) years and 61% were males. Of the total, about 86% had pulmonary TB, 95% were new cases, and 23% were microbiologically confirmed (either on sputum smear microscopy or Xpert assay). Among 2347 patients, 19% received free drugs from the National Tuberculosis Program (NTP) under the supervision of PPs while the rest purchased anti-TB drugs from the private pharmacies. The ‘successful outcomes’ (cured and treatment completed) were seen in 86.1% (95% confidence interval (CI), 84.6%-87.4%) patients and 8.6% were ‘not evaluated’. The patients who received free NTP drugs (adjusted relative risk-4.0 (95% CI: 3.1-5.0)) had a higher risk of ‘unsuccessful outcomes’ than those who availed of drugs from private pharmacies. The high ‘successful outcomes’ among TB patients treated by PPs are promising. However, higher ‘unsuccessful outcomes’ among patients on free NTP drugs need to be explored, and suitable corrective actions have to be taken.

Factors associated with culture conversion among adults treated for pulmonary extensively drug-resistant tuberculosis during 2018-2019 in the Russian Federation: an observational cohort study.

Treatment outcomes for Multidrug/Rifampicin-Resistant Tuberculosis (MDR/RR-TB) and Extensively Drug-Resistant Tuberculosis (XDR-TB) remain poor across the globe and in the Russian Federation. Treatment of XDR-TB is challenging for programmes and patients often resulting in low success rates and onward transmission of drug-resistant strains. Analysis of factors affecting culture conversion rate among XDR-TB patients may serve as a basis for optimization of treatment regimens. We conducted a retrospective cohort study using health records from 54 patients with pulmonary XDR-TB treated at a tertiary level facility in the Russian Federation. The study population included adult patients with culture-positive pulmonary XDR-TB who started treatment between 1 January 2018-30 June 2019. Culture conversion was defined as two consecutive negative cultures, collected at least 30 days apart. The date of sputum culture conversion was taken from the first of two consecutive negative sputum cultures fulfilling these criteria. We measured time to culture conversion using cumulative incidence functions accounting for competing risks and applied binary cause-specific Cox regressions to assess associated factors. Sputum culture conversion was recorded for 43 (79.6%) patients. Median time to culture conversion adjusted for competing risk of loss to follow up was 4 months [95% confidence interval (CI): 2-5]. The number of patients who had culture converted by treatment months 2, 4, and 6 were 12 (22%), 29 (54%) and 38 (70%) respectively. In unadjusted analysis, positive baseline sputum smear microscopy [hazard ratio (HR): 0.34, 95% CI: 0.18-0.66; p=0.001), hepatitis C (HR: 0.35, 95% CI: 0.14-0.89; p=0.023], and human immunodeficiency virus (HR: 0.30 95%, CI: 0.09-0.97; p=0.045), and receipt of fewer than 4 effective drugs in the treatment regimen (HR: 0.13, 95% CI: 0.03-0.60; p=0.009) were associated with delayed culture conversion. When compared to their combined use, patients receiving regimens with bedaquiline only (HR: 0.12, 95% CI: 0.03-0.49; p=0.003) orlinezolid only(HR: 0.21, 95% CI: 0.06-0.69; p=0.010) were less likely to achieve timely culture conversion. Factors delaying sputum culture conversion should be considered in the management of patients with XDR-TB and considered by clinicians for regimen design and treatment strategies. Our study outlines the importance of simultaneous inclusion of bedaquiline and linezolid in treatment regimens for patients with XDR-TB to reduce time to sputum conversion and increase treatment success.

Factors associated with time to sputum culture conversion of rifampicin-resistant tuberculosis patients in Klaipeda, Lithuania in 2016-2019: a cohort study.

The global proportion of successful treatment outcomes of Multidrug-Resistant/Rifampicin-Resistant Tuberculosis (MDR/RR-TB) remains unacceptably low. Time to culture conversion is important in making treatment-related decisions and is used as an interim predictor of pulmonary MDR/RR-TB treatment success. No previous studies have been conducted to assess determinants of time to culture conversion for MDR/RR-TB patients in Lithuania. Secondary analysis of data of culture-positive MDR/RR-TB patients, treated in Republican Klaipeda Hospital between 1st July 2016 and 1st July 2019 was performed. Culture conversion was defined as two consecutive negative cultures on solid media submitted at least 30 days apart. Factors associated with culture conversion were estimated by crude and multivariable Cox regression accounting for competing risks.In total, 115 consecutive patients starting treatment were included in the study. Of them, the majority was male (86/115; 74.8%) with a mean age of 48 (standard deviation (SD) ±12) years and Human Immunodeficiency Virus (HIV) negative (105/115; 91.3%). Nearly two-thirds (72/115; 62.6%) had XDR (extensive drug resistance) or MDR/RR-TB with additional resistance to second-line injectables or fluoroquinolones. Of 115 culture-positive patients at baseline, 103 (89.6%) patients achieved culture conversion during 12 months of treatment. The median time to culture conversion was 1.1 months (interquartile range: 0.9-1.8). Patients aged ≥60 years compared with <40 years [adjusted hazard ration (aHR): 0.40, 95% confidence interval (CI): 0.18-0.86], smokers (aHR: 0.39, 95% CI: 0.2-0.73), patients with positive sputum smear microscopy at baseline (aHR: 0.40, 95% CI: 0.25-0.63), cavities on initial chest X-ray (aHR: 0.56, 95% CI: 0.35-0.88) and resistance to at least one fluoroquinolone drug (aHR: 0.52, 95% CI: 0.32-0.84) were slower to culture convert. In conclusion, we recommend providing additional counseling, treatment adherence interventions and scale up the use of new and repurposed TB drugs to patient groups at risk of worse interim treatment outcome: patients aged 60 and above, with resistance to fluoroquinolones, smear-positive, smokers, or with signs of extensive disease evident on initial chest radiography.

How Much Do Smear-Negative Patients Really Contribute to Tuberculosis Transmissions? Re-Examining an Old Question with New Tools

Background: Sputum smear microscopy is a common surrogate for tuberculosis infectiousness. Previous estimates that smear-negative patients contribute 13-20% of transmissions and are, on average, 20-25% as infectious as smear-positive cases are understood to be high. Herein, we use an ideal real-world setting, a comprehensive dataset, and new high-resolution techniques to more accurately estimate the true transmission risk of smear-negative cases. Methods: We treated all adult culture-positive pulmonary TB patients as potential transmitters. Over a 15.5-year period we measured, as primary outcomes, the proportion of transmissions attributable to smear-negative sources and the relative transmission rate. First, we replicated previous studies by using only molecular (DNA) fingerprint clustering. Then, using a prospectively collected registry of TB contacts, we defined transmission events as active TB among identified contacts who either had a 100% DNA fingerprint match to the source case or a clinical diagnosis. To minimize omission of secondary cases who had not been identified as contacts, we undertook genome sequencing on temporally and geographically linked DNA fingerprint clusters. Findings: There were 1,176 cases, 563 smear-negative and 613 smear-positive, and 23,131 contacts. Replicating previous studies, the proportion of transmissions attributable to smear-negative source cases was 16% and the relative transmission rate was 0.19. With our combined approach, the proportion of transmission was 8% and the relative transmission rate became 0.10. Interpretation: After enhancing molecular data with conventional epidemiologic, spatial/temporal and genomic data, we found that smear-negative cases were approximately 50% less infectious than previously thought. The public health implications of this are substantial. Funding: Alberta Innovates Health Solutions Declaration of Interest: We declare no competing interests. Ethical Approval: The University of Alberta Health Research Ethics Board (HREB) Biomedical Panel for review of non-invasive studies involving humans provided approval for this study, protocol ID Pro00088408.

Factors associated with unfavorable outcome of tuberculosis treatment in people deprived of liberty: a systematic review.

to analyze factors associated with unfavorable outcome of tuberculosis treatment in people deprived of liberty. systematic review, carried out in March 2021 in seven databases, with no delimitation of period of publication. The selection process of publications and data extraction was carried out by two independent reviewers. a total of 1,448 publications was identified and nine were included in the study. Unfavorable outcome was higher among those who were men; had low level of education; were living in a rural area before detention; had longer prison time; received occasional visits; had been transferred between prisons; with no sputum smear microscopy or with a positive result at the diagnosis; with no follow-up sputum smear microscopy, previous history of tuberculosis; having both clinical forms of the disease, HIV/AIDS; alcoholics; smokers; low body weight; and self-administered treatment. Treatment default was associated with young people and death with older people. prison health managers and professionals are expected to establish mechanisms of surveillance and health actions innovation aimed at the population deprived of liberty, making efforts to reduce the unfavorable outcomes of tuberculosis treatment.

Therapeutic itinerary of people with leprosy: paths, struggles, and challenges in the search for care.

to understand how the therapeutic itineraries of people affected by leprosy are processed. this is a descriptive, qualitative study, conducted in April 2018 in Barão de Grajaú in Maranhão, with interviews in the form of narratives of seven patients who had a late leprosy diagnosis. the search for diagnosis is a major difficulty in accessing health services, resulting in a late diagnosis and, consequently, with the presence of visible deformities. It was noticed that the health units do not have a flow, nor protocols for comprehensive treatment, and these people are referred to a referral unit in another state to perform sputum smear microscopy. leprosy control actions need reformulations that seek the relationship between operational activities, epidemiological indicators and risk factors, in accordance with the real needs of each region, thus highlighting the gaps evidenced in the therapeutic itineraries.

Bleach-concentrated direct sputum smear microscopy procedure for diagnosis of pulmonary tuberculosis in poverty areas

ABSTRACT INTRODUCTION: Pulmonary tuberculosis caused by Mycobacterium tuberculosis is a serious public health problem affecting millions of people worldwide. The development of easy and low-cost diagnostic methods is crucial for disease control in rural remote and poverty areas and among vulnerable groups. OBJECTIVE: To evaluate the accuracy of laboratory methods for the diagnosis of Pulmonary tuberculosis. MATERIAL AND METHODS: Sputum samples from patients with clinical signs and symptoms were analyzed by microscopy after chemical treatment and spontaneous sedimentation and compared with methods employed routinely: direct sputum smear microscopy, culture, and GeneXpert®MTB/RIF. RESULTS: From the samples analyzed, 16% were positive by microscopy in the processed samples, 18% by both culture and Xpert®MTB/RIF, while 13% in the direct microscopy. The processed samples showed a 31% increase in positivity (57 samples) compared to conventional direct microscopy. In the analysis of the accuracy of the evaluated methods, all the results were statistically significant proving that they were not randomly positive or negative and confirming that there is a tendency for these results. CONCLUSION: Chemical treatment and spontaneous sedimentation of the sputum samples procedure represent an effective diagnostic tool in situations where more advanced technologies are not feasible. Besides the higher accuracy and greater detection of positive cases regarding the direct smear, the procedure strengthens biosafety by decreasing the risks of aerial contamination by Mycobacterium tuberculosis for laboratory professionals.

Increased meat consumption is associated with reduced tuberculosis treatment failure rate: A cohort study.

Information regarding the relationship between dietary meat intake and tuberculosis treatment outcomes among patients with tuberculosis is still limited. The aim of the present study was to investigate the relationship between meat consumption and tuberculosis treatment outcomes in patients with pulmonary tuberculosis. A pulmonary tuberculosis cohort study including 2,261 patients with pulmonary tuberculosis was conducted in Linyi, Shandong Province, China from 2009 to 2013. Dietary data were collected using a semi-quantitative food frequency questionnaire. Treatment outcomes were assessed using a combination of sputum smear microscopy and chest computerized tomography. In a multiple adjusted model, the higher quartile of total meat (OR=0.59, 95% CI, 0.38-0.91) was associated with a decreased failure rate of pulmonary tuberculosis treatment. In addition, higher consumption of chicken (OR=0.62; 95% CI, 0.44-0.87) and pork (OR=0.54; 95% CI, 0.31-0.95) was associated with a lower failure rate of tuberculosis treatment. Increased dietary intake of total meat, pork, and chicken are associated with a decreased failure rate of pulmonary tuberculosis treatment. A higher dietary meat intake may be beneficial in pulmonary tuberculosis treatment.

Study of quality of sputum being submitted for smear examination

Background: One of the key factors influencing the results of sputum smear microscopy is the quality of sputum being submitted. Methods: This is a prospective study of quality of sputum specimens being submitted for smear examination under Revised National Tuberculosis Control Programme and its impact on diagnostic yield of sputum smear microscopy. Based on sputum Gram's staining results, the submitted specimen was classified as 'sputum' and 'saliva'. Results: During the period 21 April to 20 June 2012, 238 sputum specimens submitted for evaluation were studied. Of these, 196 (82%) specimens were submitted for diagnosis and 42 (18%) specimens were obtained during follow-up evaluation. Overall, 42/238 (17.6%) specimens were in fact 'saliva' and not sputum. Among specimens submitted for diagnosis (n = 196), 28/167 (16.8%) specimens reported as 'sputum acid-fast bacilli (AFB) negative' were in fact saliva. Among the specimens submitted on follow-up examination (n = 42), 14 of the 38 (36.8%) specimens reported as 'sputum AFB negative' were actually saliva. A significantly higher proportion of sputum smears submitted on follow-up examination were actually 'saliva' as compared to specimens submitted for diagnosis (14/42 [33.3%] vs. 28/196 [14.3%], χ2 = 6.738; P = 0.014). None of the saliva samples were reported to be 'sputum positive'. Conclusions: The fact that a significant proportion of specimens that are reported smear negative are in fact saliva and not sputum highlights the importance of sputum quality in smear microscopy results. Follow-up specimens are more likely to be saliva possibly because directly observed therapy (DOTS) treatment would have resulted in clinical improvements and patients may not be producing significant sputum anymore.

Molecular bacterial load assay versus culture for monitoring treatment response in adults with tuberculosis.

The lack of rapid, sensitive, and deployable tuberculosis diagnostic tools is hampering the early diagnosis of tuberculosis and early detection of treatment failures. The conventional sputum smear microscopy or Xpert MTB/RIF assay cannot distinguish between alive and dead bacilli and the culture method delays providing results. Tuberculosis molecular bacterial load assay is a reverse transcriptase real-time quantitative polymerase chain reaction that quantifies viable tuberculosis bacillary load as a marker of treatment response for patients on anti-tuberculosis therapy. However, results are not synthesized enough to inform its comparative advantage to tuberculosis culture technique which is yet the gold standard of care. With this review, we searched electronic databases, including PubMed, Embase, and Web of Science, from March 2011 up to February 2021 for clinical trials or prospective cohort studies that compared tuberculosis molecular bacterial load assay with tuberculosis culture in adults. We included eight studies that meet the inclusion criteria. Tuberculosis molecular bacterial load assay surpasses culture in monitoring patients with tuberculosis during the first few weeks of anti-tuberculosis treatment. It is more desirable over culture for its shorter time to results, almost zero rates of contamination, need for less expertise on the method, early rate of decline, lower running cost, and reproducibility. Its rapid and specific tuberculosis treatment monitoring competency benefits patients and healthcare providers to monitor changes of bacillary load among isolates with drug-susceptible or resistance to anti-tuberculosis regimens. Despite of the high installing cost of the tuberculosis molecular bacterial load assay method, molecular expertise, and a well-equipped laboratory, tuberculosis molecular bacterial load assay is a cost-effective method with comparison to culture in operational running. To achieve maximum utility in high tuberculosis burden settings, an intensive initial investment in nucleic acid extraction and polymerase chain reaction equipment, training in procedures, and streamlining laboratory supply procurement systems are crucial. More evidence is needed to demonstrate the potential large-scale and sustainable use of tuberculosis molecular bacterial load assay over culture in resource-constrained settings.

Performance of Loop-Mediated Isothermal Amplification (LAMP)-Lateral Flow Detection and Xpert MTB/RIF Ultra for Diagnosis of Pulmonary Tuberculosis

Background: Due to the limitations of diagnosis by Xpert MTB/RIF assay, WHO suggests Loop—Mediated Isothermal Amplification (TB-LAMP) instead of sputum-smear microscopy for pulmonary TB diagnosis in patients. Dr. Thongchai Kaewphinit et al. invented a fast TB detection kit called TB d-tect (LAMP-LFD assay). There was no clinical trial to estimate the performance of TB d-tect. Objective: This study was aimed to find the performance of LFD assay and Xpert MTB/RIF ultra for pulmonary TB. Material and methods: A cross-sectional study was conducted. Suggestive pulmonary TB patients were enrolled from June 2020-28 February 2021. Respiratory specimens were collected from each patient and sent for AFB smear, LAMP-LFD assay, Xpert MTB/RIF ultra and culture TB. Result: 139 patients with suspected pulmonary TB were enrolled. 51% of patients were diagnosed with pulmonary tuberculosis. Based on culture TB as a gold standard, the sensitivity and specificity of LAMP-LFD assay were 85.4% (95% CI: 70.8% - 94.4%) and 87.8% (95% CI: 79.6 - 93.5), respectively. The sensitivity and specificity of Xpert MTB/RIF ultra were 95.1% (95% CI: 83.5% - 99.4%) and 74.5% (95% CI: 64.7 - 82.8), respectively. According to ROC curve, it was found that the areas under the curve of LAMP-LFD assay and Xpert MTB/RIF ultra were 0.866 and 0.848, respectively (p = 0.546). Conclusion: The diagnostic sensitivity and specificity of LAMP-LFD assay appeared to be comparable to those of Xpert MTB/RIF ultra. LAMP-LFD assay could be used in resource limiting laboratory.

A systematic review of utility of urine lipoarabinomannan in detecting tuberculosis among HIV-positive tuberculosis suspects.

Sputum smear microscopy (SSM), though regarded as an inexpensive and popular method for detecting tuberculosis (TB), lacks adequate sensitivity, specifically in adult people living with HIV/AIDS (PLHIV). Urine lipoarabinomannan (LAM) is a promising diagnostic tool among PLHIV with CD4 cell count < 200 cells/μl. We attempted to review all the studies undertaken in identifying the utility of urine LAM in diagnosing TB, especially among PLHIV. We searched PubMed, Google Scholar, and MEDLINE databases for studies reporting diagnostic utility of urine LAM status in PLHIV, published in the last 20 years till December 2019. The keywords used for searching were “Tuberculosis,” “HIV/AIDS,” “Diagnosis,” “Screening” “Lipoarabinomannan,” and “Urine.” Our search resulted in 137 shortlisted citations, of which 67 related manuscripts were identified for detailed study. Based on inclusion and exclusion criteria, 37 studies were reviewed in detail. Average sample size of these studies was 464 (range = 81–2528; SD = 427). Crude average sensitivity of urine LAM in culture-confirmed TB cases was 44.1% (range = 8.3–93) while that of SSM was 38.6% (range = 14–65). However, sensitivity of urine LAM + SSM was 60.4% (range = 38.3–92.7), demonstrating the utility of SSM + urine LAM combination for detecting TB. Specificity was similar between urine LAM and SSM with 92.7% (range = 76–100) and 97.9% (range = 93.9–100), respectively. Majority of the studies demonstrated higher sensitivity of urine LAM in those with lesser the CD4 count, with immunocompromised and with debilitation who cannot produce self-expectorated sputum. We conclude that urine LAM is a potential diagnostic test in the algorithms involving immunocompromised, debilitated patients and specifically in PLHIV whose CD4 count is ≤100 cells/μl.

Comparison of staining techniques– Ziehl Neelsen stain, Gabbet’s Stain, Fluorochrome stain for detection of mycobacterium tuberculosis in sputum

In developing countries, Ziehl-Neelsen (Z-N) sputum smear microscopy is widely used.The present prospective study was under taken to see the efficacy of Ziehl-Neelson method versus Gabbett and Fluorescent staining in the detection of mycobacteria in sputum sample. 80 sputum samples were collected from suspected tuberculosis patients attending RNTCP OPD at, Nerul, Navi Mumbai. Triplicate smears were prepared from all these samples and stained by the Z-N, Gabbet’s, and Fluorescent stain. Number of smears positive for Acid Fast Bacilli by, Z-N method 13(16%), Gabbett’s method 7(9%), Fluorescent method 26(33%). Combination of two staining methods yielded following results: Z- N + Fluorescent26 (33%) and Z- N + Gabbett 13(16%). The conclusion from this study is that when smear examination by Z-N is done, there are possibilities of false negative results which can be reduced by using Fluorescent staining. The rate of positivity can be improved by combination of Z-N + Fluorescent staining. So, it may be concluded that combination of ZN and Fluorescent Microscopy may be considered for diagnosis of pulmonary tuberculosis in any diagnostic set up along with culture which is the gold standard. Keywords: Fluorescent, Gabbett’s, Mycobacterium tuberculosis, Revised National Tuberculosis Control Programme (RNTCP), Zehil-Neelson (Z-N).

Risk factors for multiple drug resistant tuberculosis relapses

The objective of the study: to identify risk factors for relapses in patients after the effective course of chemotherapy for multiple drug resistant tuberculosis (MDR TB). Subjects and methods. Medical files of 346 adult MDR TB patients were analyzed, they all had the effective treatment as per regimen IV in Tomsk Phthisiopulmonology Medical Center in 2009-2011. They were divided into 2 groups: Group 1 included 35 patients who developed a relapse of tuberculosis over the next 5 years; Group 2 consisted of 311 patients who had no relapse of the disease over the next 5 years. Results. The relapse rate in effectively treated MDR TB patients made 10.1% (35/346 patients). The following social factors contributed to higher chances of the relapse: retirement age (p = 0.045; OR = 2.86 [1.14-7.20]) and disability due to somatic diseases (p &lt; 0.001; OR = 4.82 [2.13-10.90]); while biomedical factors were the following: HIV infection (OR = 19.19 [5.29-69.56]), mental illness (OR = 5.85 [2.27-15.03]), tobacco smoking (OR = 3.16 [1.08-9.20]). People with history of tuberculosis relapses (OR = 12.17 [4.19-35.34]) have higher chances of relapse, as well as those having the following characteristics of the tuberculosis disease during the effective chemotherapy course: destruction of lung tissue (OR = 7.48 [1.76-31.80]), positive results of sputum smear microscopy (OR = 2.91 [1.28-6.61]), persisting bacterial excretion (by culture) after 2 months of chemotherapy (OR = 4.98 [2.41-10.29]).

Positivity Rate of Pyrosequencing to Diagnose Drug-Resistant Tuberculosis Directly from Sputum with Different Bacterial Load

BACKGROUND: Molecular techniques, which detect mutations associated with drug resistance tuberculosis (TB), are promising technologies for rapid diagnosis and monitoring of drug-resistant TB. Pyrosequencing is a potential rapid and robust molecular technique to detect drug resistance but its performance in clinical samples is less investigated. This study aimed to determine the positivity rate of pyrosequencing to diagnose drug-resistant TB directly from sputum samples with different grades of sputum smear microscopy results.METHODS: Thirty-five sputum specimens from drug-resistant TB suspects were submitted for acid-fast bacilli (AFB) microscopy. All specimens were cultured using microscopic observation drug susceptibility (MODS) culture. Pyrosequencing was performed to DNA extracted from sputum of culture-positive patients.RESULTS: MODS culture was positive in 19/35 subjects (54.29%) samples; 16 smear-positive and three smear-negative. Using pyrosequencing, Mycobacterium tuberculosis was identified in all culture-positive samples, including smear-negative samples. A complete resistance profile for 16 (82.35%) samples could be generated. Pyrosequencing failed to show results for eis or gyrA promoter in three samples. Nine of 19 patients were multidrug resistant-TB (MDR-TB), 1/19 was rifampicin-resistance TB (RR-TB), and 4/19 were pre-extensively drug-resistant TB (pre-XDR-TB). Two novel mutations in rpoB and rrs (associated with rifampicin and aminoglycoside, respectively) were found in this study.CONCLUSION: The results of this study demonstrates high positivity rates of pyrosequencing to detect drug-resistant TB directly from sputum samples with different grades of smear microscopy, as the surrogate of bacterial load. The assay can be used as a first prediction test of drug resistance prior to confirmation by phenotypic tests.KEYWORDS: drug-resistant tuberculosis, pyrosequencing, direct sputum

Retrospective analysis of rifampicin-resistant pattern of Mycobacteria tuberculosis in secondary referral hospital Offa, Nigeria

Background: Multidrug-resistant tuberculosis as Pulmonary Tuberculosis (TB) caused by Mycobacteria tuberculosis is the leading cause of death among infectious agents and has constituted a threat to global health security. Despite major efforts to increase case detection, an estimated one-third of new tuberculosis cases are still being missed each year, due to unavailability of a rapid, low-cost, accurate diagnostic method as Nigeria health systems still rely heavily on out-dated tuberculosis diagnostic tests; including sputum smear microscopy and chest radiography. Methods and materials: This sought to conduct a retrospective review of presumptive TB register of patients screened for Mycobacterium tuberculosis (MTB) and rifampicin-resistant tuberculosis (RR-TB) using Xpert MTB/RIF assay at General Hospital Offa in Kwara State Nigeria between January to December 2018 in order to access the on-going efforts such as quarterly surveillance, improved infrastructural facilities and intense media advocacy on Tuberculosis by Kwara state government on prevention of Tuberculosis in Kwara State. Results: Overall, 63 (10.6%) of the sputum specimens were positive for Mycobacterium tuberculosis while 4 (6.3%) showed rifampicin resistance out 63 (10%) of patients positive for Mycobacteria tuberculosis. There is a high recovery rate of Mycobacteria tuberculosis among women than men as this study recovers 52.4% and 47.6% in male and female respectively. This study documents low frequency of rifampicin-resistant tuberculosis particularly within the female population using highly sensitive and specific Xpert MTB/RIF in Kwara State. Conclusion: Use of validated molecular method for MTB strain detection has proven the stronger epidemiological impetus for an enhanced care management outcome especially in resources limited regions.