GeneXpert MTB Research Articles

A CLINICO-PATHOLOGICAL STUDY ON CYTOLOGICAL SCREENING OF BRONCHOALVEOLAR LAVAGE AND SUBSEQUENT CORRELATION WITH ADENOSINE DEAMINASE, GENEXPERT MTB IN SUSPECTED SPUTUM NEGATIVE PULMONARY TUBERCULOSIS

BACKGROUND: Sputum negative pulmonary tuberculosis is a world-wide burden on health budget. Clinico- radiologically suspected sputum negative pulmonary tuberculosis needs to be diagnosed using cytopathology along with ADA levels and GeneXpert MTB tests of BAL fluid obtained through bronchoscopy. AIM: To interpret the cytological screening of bronchoalveolar lavage and subsequent correlation with adenosine deaminase, genexpert mtb in suspected sputum negative pulmonary tuberculosis. METHODS: Patients visiting Pulmonary Medicine outdoor of R.GKAR Medical College Hospital, who are clinico radiologically being suspected as pulmonary tuberculosis but are sputum smear AFB negative on two consecutive samples. The study group includes these patients who are willing to undergo bronchoscopy, in whom there are neither any medical contraindications nor other miscellaneous restraints i.e. the study group contains cases of pulmonary TB as well as other infectious malignant non-infectious lung diseases. RESULT: Of the 61 patients, 41(67.2%) patients were male and 20(32.8%) patients were female. The average age of male patients was 51.71years and the average age of female patients was 39.35years.The mean ADA level of BAL fluid in patients with Pulmonary Tuberculosis was 22.54 U/L (Normal level – less than 30 U/L) and mean ADA level of BAL fluid in patients without Pulmonary Tuberculosis was 4.76 U/L (Normal level – less than 30 U/L). CONCLUSION: Cytology of bronchoalveolar lavage along with adenosine deaminase and GeneXpert MTB are important diagnostic and screening tools in suspected sputum negative pulmonary tuberculosis. As it can detect different kinds of inflammatory and other types of cells, it can also help in choosing the appropriate therapeutic procedure / modality for a disease.

Evaluation of Clinical background and yield of chest tube thoracotomy in hydropneumothorax , a pulmonologist point of view.

Introduction: The enlightenment of Hydro-pneumothorax arrived in former times of primitive Greece. It is a term which explains simultaneous existence of both free air and fluid (i.e., pneumothorax and hydrothorax) in the pleural space.The goal was to contribute and update the knowledge of clinical aspect as well as yield of diagnostic modalities in handling the cases of Hydropneumothorax. Materials and Methods: Diagnosed cases of hydropneumothorax from casualty and routine OPD are considered. Detail of clinical background entered. Patients were subjected for analysis of sputum, blood and pleural fluid. Radiological investigation was done. Patients underwent tube thoracotomy procedure and observed till resolution. Results: Total 97 subjects had participated. Among them 76(78.35%) were men and 21 (21.64%) women. 91(93.85%) subjects had shortness of breath. 34 subjects (35.05%) had tuberculosis in the past. Sputum smear AFB was positive in 19 subjects (19.58%). Plural fluid GeneXpert MTB was detected in 11 subjects (11.34 %) and MGIT Culture positive in 23(23.71%) subjects. Plural fluid gram stain culture was conclusive in 24 (24.74%) subjects. Pleural fluid ADA was raised in 74 (76.28%) subjects. Hypoxemia was observed in 46 (47.42%). In 38 (39.19%) cases chest tube was removed between 31 to 60 days. Conclusion: Tuberculosis turns up as the most common aetiology of hydropneumothorax and duration required for resolution is uncertain. Evaluation of pleural fluid (cytology, microbiological culture, biochemical measure) and Computed tomography were essential to reach aetiology of condition. Chest tube thoracotomy remains most practiced modality of treatment in hydropneumothorax. However advanced techniques have promising outcome and it’s an opportunity to research further.

Lobektomi Life Saving pada Hemoptisis Berulang Pada Tuberkulosis Paru

Background. Hemoptysis is the most common complication of lung tuberculosis (TB). Its can occur repeated and massive. Reccurent massive hemoptysis resolved with conservative therapy but if conservative therapy fails, definitive therapies such as lobectomy surgery are used as alternative treatments especially life-threatening. Case. A 38-year-old man with chief complaint hemoptysis since 4 months pior to admission hospital. Hemoptysis 4-5 times daily with volume 200-600 cc per day and make him shortness of breath and anemia. Initial AFB (Acid Fast Bacilli) was positive and GeneXpert MTB (+) sensitive rifampisin. He got Anti Tuberculosis Drug (ATD) therapy 1st category since 3 months ago. He has been given conservative therapy for hemoptysis but failed. Discussion. Surgery is one of alternative therapy for reccurent massive hemoptysis. Surgery only if source of bleeding already known certainty. In this case patient were suggest for lobectomy superior right lung lobe, not a segmentectomy because there was adhesions on intrathoracic. After lobectomy he getting better and hemoptysis stops. He has continued ATD until 6 months without hemoptysis. Conclusion. Reccurent massive hemoptysis and life-threatening with failure of conservative therapy may given definitive therapy such as lobectomy

Evaluation of GeneXpert MTB for detection of Mycobacterium tuberculosis in Fayoum Governorate

Background Despite being one of the most ancient diseases, tuberculosis (TB) is still a major health problem globally. It is one of the most important causes of death worldwide. GeneXpert MTB/RIF assay is a fast nucleic acid amplification test used for detection of TB as well as identification of rifampicin resistance. Design This was a cross-sectional study. Setting The study was conducted at Fayoum Chest Hospital, Egypt, between December 2017 and December 2018. Objectives To evaluate GeneXpert MTB/RIF in detection of Mycobacterium tuberculosis in Fayoum Chest Hospital. Patients and methods One hundred participants were enrolled in this study with clinical or radiological susceptibility of pulmonary or extrapulmonary TB. All patients underwent the following: full history taking, clinical examination, plain chest radiography, examination of specimen by Ziehl–Neelsen stain, cultivation on Lowenstein–Jensen media, and processed in GeneXpert MTB/RIF. Results GeneXpert assay is a good, fast, noninvasive, and accurate diagnostic tool for detection of M. tuberculosis, showing sensitivity of 100.0%, specificity of 85.0%, positive predictive value of 96.4%, negative predictive value of 100%, and accuracy of 97%. On the contrary, Ziehl–Neelsen had sensitivity of 83.75%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 60.61%. Conclusion GeneXpert is a fast, accurate, and promising technique with good sensitivity and specificity, but it cannot be used alone as a diagnostic tool for TB.

GeneXpert's Performance in the Diagnosis of Pulmonary and Extrapulmonary Tuberculosis

The diagnosis of tuberculosis is based on the isolation of acid-fast bacilli (AFB) on direct examination or on the culture isolation of mycobacteria tuberculosis complex (MCT). Currently, the introduction of molecular biology, essentially the Xpert / MTB / Rif or GeneXpertR test has increased the sensitivity and speed of diagnosis. The objective of our study is the evaluation of the performance of the Xpert / MTB / Rif molecular technique in the diagnosis of pulmonary and extrapulmonary tuberculosis. This is a descriptive retrospective study over a period of 22 months (June 2016 - April 2018). The study concerned all pulmonary and extrapulmonary specimens sent to the laboratory of bacteriology-virology and molecular biology of the HMA Marrakech for suspicion of tuberculosis. Diagnosis was performed on all specimens by direct examination on Ziehl-Neelsen stained smears, culture on Loweinstein Jensen solid medium (LJ) and GeneXpert molecular technical. Our study included 305 patients, 69% were men and 31% women. The clinical samples included in the study consisted of different specimens with a prevalenceof sputum (n = 127). Positivity was higher for GeneXpert MTB / RIF than for Ziehl-Neelsen and culture with respectively 18% (n = 54) for GeneXpert against 13% (n = 40) for culture and 8% (n = 24) for Ziehl-Neelsen. On all positive direct examination specimens, GeneXpert was positive in 100% of cases, on negative direct examination samples, GeneXpert was positive in 5% of cases. The positivity rate of GeneXpert was 10.6% for samples with negative culture and 100% for positive culture samples. The performance of GeneXpert was variable depending on the nature of the samples: sputum (23%), biopsy (15%), pleural fluid (13%). Four cases of resistance to rifampicin have been detected. Our study shows the great contribution of GeneXpert for the diagnosis of pulmonary and extrapulmonary tuberculosis as well as its place in the determination of multidrug resistance.

The use of Gene-Xpert MTB RIF in the diagnosis of extrapulmonary tuberculosis in childhood and adolescence.

INTRODUCTION: Gene-Xpert MTB RIF (Xpert) is based on nucleic acid amplification by real-time polymerase chain reaction, which allows for the identification of Mycobacterium tuberculosis and rifampin resistance. We describe the use of Xpert for extrapulmonary tuberculosis (EPTB) in children and adolescents. METHODS: A case series of two reference centers in Rio de Janeiro from 2014-2019. RESULTS: The final diagnosis of EPTB was established in 11/36 (31%) patients, with five cases detectable by Xpert. For lymph node evaluation (9/11), diagnosis by Xpert occurred in 5/9 patients, all with caseous aspects. CONCLUSIONS: Xpert can facilitate the rapid diagnosis of lymph node tuberculosis.

Comparison of Led Fluorescent Microscopy and the Gene Xpert MTB/RIF Assay in Diagnosis of Pulmonary and Extrapulmonary Tuberculosis

Objectives: The objective of this study was to evaluate Gene Xpert MTB/RIF Assay and anid fast staining (AFB) for rapid detection of Mycobacterium tuberculosis in specimen of patients suspected of pulmonary tuberculosis (PTB) and extra pulmonary tuberculosis (EPTB).
 Methods: A comparative cross-sectional study of 400 samples (PTB-365 and EPTB-35) of patients visiting National Tuberculosis Centre (NTC) was conducted from July 2018 to December 2018. Gene Xpert MTB/ RIF Assay, smear microscopy were performed under standard guideline inside biosafety cabinet class II. The result obtained from both the tests were analyzed using SPSS 20.0 software and Excel 2019.
 Results: Of the total samples, 18% (72/400) and 39% (156/400) were positive by AFB smear microscopy and Xpert MTB/RIF assay respectively. Prevalence of MTB positive was highest in the age group 35-44 years, 33 cases (17.74%) were detected in total, with a male to female ratio of 2.3:1. Pleural fluid, pus, and CSF fluid also yielded positive results with the Gene Xpert MTB/RIF assay accounting 1.28%, 0.64% and 1.28% of MTB positive case respectively. Rifampicin resistance was observed in 1.28% of the cases.
 Conclusion: The key findings of this study suggest that Gene Xpert test should be implemented as primary diagnostic test for PTB and EPTB.

EFFICACY OF GENE XPERT MTB/RIF ASSAY AND AFB CULTURE IN DIAGNOSIS OF MYCOBACTERIUM TB COMPLEX ON EBUS TBNA

SESSION TITLE: Tuesday Abstract Posters SESSION TYPE: Original Investigation Posters PRESENTED ON: 10/22/2019 01:00 PM - 02:00 PM PURPOSE: Rationale – Tuberculosis and Sarcoidosis are two clinical entity that share same clinical, radiological and pathological features and differentiating between two might be an arduous task. As cultures for M. Tuberculosis in tissue specimens have prolonged turn-around times, rapid molecular methods such as MTB. Rif system has shown great promise in rapid diagnosis of tuberculosis through detection of DNA of M. Tuberculosis and concurrent identification of major mutation that confers rifampicin resistance. We aim to evaluate both efficacy and role of Xpert MTB / RIF Assay and Mycobacterium TB Culture for diagnosis of TB using EBUS TBNA samples in patients with intra-thoracic granulomatous lymphadenopathy. METHODS: This was a retrospective study of randomly selected 100 patients who underwent EBUS TBNA for intra-thoracic granulomatous lymphadenitis with Gene Xpert MTB / RIF Assay and diagnosed with either Tuberculosis or Sarcoidosis RESULTS: EBUS TBNA procedures were performed and a diagnosis of Sarcoidosis (n 17) or Tuberculosis (n 83) was made in 100 patients. Histological results with grades I to II were seen in 74 patients and grade III in 26 patients respectively. Necrotising granulomas were found in 26 / 83 (31.32%) while Non-necrotising granulomas were found in 57 / 83 (68.67%) patients diagnosed with Tuberculosis. Mycobacterium TB Culture was performed in 72 patients diagnosed with Tuberculosis and was positive in 11 patients – 09 / 26 Necrotising Granuloma vs 02 / 57 Non-necrotising Granuloma (34.61% vs 3.5%, P = 0.010). Gene Xpert MTB / RIF Assay were performed in 20 / 83 patients diagnosed with Tuberculosis and was positive in 4 patients – 01 / 07 Necrotising Granuloma vs 03 / 13 Non-necrotising Granuloma (14.30% vs 23.10%, P = 0.547). The Diagnostic Accuracy of Xpert MTB / RIF Assay in diagnosis of Tuberculosis was 70% (95% CI 45.72% to 88.10%), Sensitivity 40%, Specificity 100%, Disease Prevalence 50%, PPV 100%, NPV 62.50%. Of 2 patients finally diagnosed with Sarcoidosis also showed a positive result of Xpert MTB / RIF Assay which possibly suggests casual link between TB and Sarcoidosis CONCLUSIONS: Gene Xpert MTB / RIF Assay can be useful in patients with suspected Pulmonary TB either AFB smear negative or positive due to its rapid and simultaneous detection of Rifampicin resistance and especially beneficial in patients with MDR TB and HIV associated TB. Xpert MTB / RIF Assay has good sensitivity and PPV in context of culture positive intra-thoracic TB lymphadenopathy and can provide immediate diagnosis of likely drug resistance specially in highly endemic area like India CLINICAL IMPLICATIONS: Positive Xpert MTB / RIF Assay with very low MTB detection but culture negative results need to be read cautiously and should be well correlated with clinical and treatment history of patient since Gene Xpert Assay amplifies any DNA, of live or dead bacilli and required to avoid false positive result DISCLOSURES: No relevant relationships by UMANG SHAH, source=Web Response

DIAGNOSTIC ACCURACY OF XPERTMTB/RIF ASSAY IN DETECTION OF PULMONARY TUBERCULOSIS AND RIFAMPICIN RESISTANCE IN COMPARISON WITH CONVENTIONAL METHODS..

Introduction: Tuberculosis is one of the oldest diseases with high morbidity and mortality. India accounts for one- fourth of the global TB burden (2.7million cases). Material and methods: A total 1056 patients were included in this study. All patients were requested to give two sputum samples, spot sputum sample (at the time of visit) and early morning sputum for mycobacterial testing. All spot samples tested with smear microscopy and Xpert MTB/Rif assay. Rifampicin resistant samples compared with conventional method. Results: Out of 1056 sputum samples GeneXpert MTB Rif assay detected positive 204 (19.3%), not detected 827 (78.31%). There is significant difference founded for positivity in spot samples and early morning sputum samples (P =0.0026). Rifampicins resistant were 8 (3.9%) andshown 100% sensitivity, specificity with Conventional method. Conclusion: For diagnosing tuberculosis and detecting Rifampicin resistance GeneXpert MTB/RIF has been especially recommended.

DIAGNOSTIC ACCURACY OF XPERTMTB/RIF ASSAY IN DETECTION OF PULMONARY TUBERCULOSIS AND RIFAMPICIN RESISTANCE IN COMPARISON WITH CONVENTIONAL METHODS.

Introduction: Tuberculosis is one of the oldest diseases with high morbidity and mortality. India accounts for one- fourth of the global TB burden (2.7million cases). Material and methods: A total 1056 patients were included in this study. All patients were requested to give two sputum samples, spot sputum sample (at the time of visit) and early morning sputum for mycobacterial testing. All spot samples tested with smear microscopy and Xpert MTB/Rif assay. Rifampicin resistant samples compared with conventional method. Results: Out of 1056 sputum samples GeneXpert MTB Rif assay detected positive 204 (19.3%), not detected 827 (78.31%). There is significant difference founded for positivity in spot samples and early morning sputum samples (P =0.0026). Rifampicins resistant were 8 (3.9%) andshown 100% sensitivity, specificity with Conventional method. Conclusion: For diagnosing tuberculosis and detecting Rifampicin resistance GeneXpert MTB/RIF has been especially recommended.

Bacteriologically confirmed extra pulmonary tuberculosis and treatment outcome of patients consulted and treated under program conditions in the littoral region of Cameroon

BackgroundExtra-pulmonary tuberculosis (EPTB) is defined as any bacteriologically confirmed or clinically diagnosed case of TB involving organs other than the lungs. It is frequently a diagnostic and therapeutic challenge with paucity of data available. The aim of this study was to assess the prevalence of bacteriologically confirmed EPTB; to determine the most affected organs and to evaluate the therapeutic outcome of EPTB patients treated under program conditions in the littoral region of Cameroon.MethodsA descriptive cross-sectional laboratory-based epidemiological survey was conducted from January 2016 to December 2017 and 109 specimens from 15 of the 39 diagnosis and treatment centers in the littoral region were obtained.Two diagnostic methods (Gene Xpert MTB and culture (LJ and MGIT) were used for EPTB diagnosis. Determine HIV1/2 and SD Biolinewere used for HIV diagnosis. Confirmed EPTB cases were treated following the national tuberculosis guide.ResultsThe prevalence of bacteriologically confirmed EPTB was 41.3% (45). All 45 cases were sensitive to rifampicin. Males were predominately more infected [26 (57.8%)] likewise the age group 31–45 years with 15 (33.3%) cases. The overall prevalence for HIV was 33.6% (36). HIV infection was present in 28.9% (13) of patients with EPTB. The most affected sites with EPTB were: Lymph nodes (66.5%), pleural cavity (15.6%), abdominal organs (11.1%), neuromeningeal (2.2%), joints (2.2%) and heart (2.2%). Overall, 84.4% of the study participants had a therapeutic success with males responding better 57.9% (p = 0.442). Therapeutic success was better (71.7%) in HIV negative EPTB patients (p = 0.787).ConclusionThe prevalence of bacteriologically confirmed EPTB patients treated under program conditions in the littoral region of Cameroon is high with a therapeutic success of 84.4% and the lymph nodes is the most affected site.

Missed pulmonary tuberculosis: a cross sectional study in the general medical inpatient wards of a large referral hospital in Ethiopia

BackgroundEvery year around 4 million people with tuberculosis (TB) are not detected. Thus may not get the medical care that they need and deserve from their respective health systems. Ethiopia is included in the 12 countries who contribute 75% of the globally estimated “missed” cases. This study assessed if there are missed Pulmonary TB (PTB) cases among inpatients of a large referral hospital in Ethiopia.MethodA cross sectional survey was conducted in the general medical wards of the large referral hospital from June to August 2015. Inpatients not diagnosed with TB were screened for TB symptoms and requested to submit a morning sputum sample for smear microscopy and molecular testing by GeneXpert MTB/RIF assay. The results of the symptom screening, smear and GeneXpert testing were analyzed as the main outcome characteristics for “missed” PTB cases.ResultOver the 3-month period, 300 inpatients were evaluated for TB. The patients median age was 38 years (IQR 26–51.5), 41% were female, median reported duration of sickness before admission was 30 days (IQR 14–240), and median body mass index (BMI) was 21.5 (IQR 20–22.67). HIV status was documented for 198/300 (66%) of patients, 37 (18.7%) were found to be HIV positive, with a median CD4 count of 176 (IQR 52–400). All 300 inpatients submitted a sputum sample and 10 (3.3%) were found to be GeneXpert MTB positive, with 4/10 also being smear positive. All GeneXpert positive inpatients reported having a cough of > 2 weeks duration. Eight had at least 3 common symptoms of TB (i.e. cough, fever, weight loss or night sweat). Co-morbidity with Diabetes Mellitus (DM) and HIV was found in 1/10 and 4/10 cases respectively.ConclusionBacteriological confirmed TB cases were found to have been “missed” amongst the general medical ward inpatients in the hospital. The identified TB cases all reported typical signs and symptoms of TB. Basic clinical practices were not being followed (i.e. history taking/documentation and requesting of appropriate laboratory tests) by the attending health care workers (HCWs) in the hospital. The index of suspicion for TB disease needs to improve and the use of more sensitive technologies, such as GeneXpert could assist the diagnosis of TB. However, the findings of the study need to be investigated in other hospital settings in Ethiopia.

Multidrug-Resistant Tuberculosis Disease in North-Kivu Province, Democratic Republic of Congo

Introduction: The emergency of Mycobacterium tuberculosis resistant to the first line drug reduced access possibility to second line drugs for appropriate treatment and required for urgent action especially in le Democratic Republic of Congo (DRC), which counts among the highest tuberculosis (TB) burden countries in Africa. Objective: To present prevalence and describe multidrug-resistant tuberculosis cases in North-Kivu Province identified by using Genexpert technology. Methods: We conducted an observational prospective study on multidrug-resistant tuberculosis (MDR-TB) cases in North-Kivu Province, DRC from 2017 to 2018. All cases of MDR-TB identified by Genexpert MTB/ RIB were included in this series. Result: Of 15,544 tuberculosis cases registered during the study period, 19 cases of MDR-TB were identified. 57.9% was male, 89.5% was retreatment cases and 5.3% was coinfection HIV/TB cases. Conclusion: This new molecular technology diagnostic facilitates multidrug-resistance tuberculosis detection and improves the reporting of data lack.

Assessment of GeneXpert MTB/RIF Performance by Type and Level of Health-Care Facilities in Nigeria.

Setting:Nigeria adopted GeneXpert MTB Rif as a primary diagnostic tool were available and accessible since 2016. The current geographical coverage of GeneXpert machines by LGAs stands at 48%, with a varied access and utilization.Objectives:To assess the association between the type and level of health facilities implementing GeneXpert MTB/Rif and performance outcome of the machines in Nigeria.Study Design:Retrospective secondary data analysis of GeneXpert performance for 2017 from GXAlert database. The independent variables were type and levels of health care facilities, and dependent variables were GeneXpert performance (utilization, successful test, error rates, MTB detected, and Rifampicin resistance detected).Results:Only 366 health care facilities are currently implementing and reporting GeneXpert performance, the distribution is 86.9% and 13.1% public and private health care facilities respectively, and only 6.3% of the facilities are primary health care. Of 354,321 test conducted in 2017, 91.5% were successful, and among unsuccessful test 6.8% were errors. The yield was 16.8% MTB detected (54,713) among which 6.8% had Rif resistance. The GeneXpert utilization rate was higher among private health care facilities (55.8%) compared to 33.3% among public health care facilities. There was a statistically significant difference in the number of successful test between public and private health facility-based machines as determined by one-way ANOVA (F(1,2) = 21.81, P = 0.02) and between primary, secondary and tertiary level health facility-based machines (F(1,2) = 41.24, P < 0.01).Conclusion:Nigeria with very low TB coverage should rapidly scale-up and decentralize GeneXpert services to the private sector.

Coinfection with Staphylococcus aureus community-acquired pneumonia and mycobacterium turberculosis in a diabetic patient: a case report

Pulmonary tuberculosis is a common reason for hospital admission worldwide. The objective of this case report is to demonstrate the importance of the active search for M. tuberculosis along with Community-Acquired Pneumonia (CAP). The 36-yearold male patient, known as diabetic, hypertensive and obese, was admitted to the emergency department of the San Carlos Hospital with bronchopneumonia, mild respiratory distress and simple diabetic decompensation. Based on the radiological findings, the patient is treated with a presumptive diagnosis of a right basal CAP and pulmonary tuberculosis. The first smear analyzed by smear microscopy was found positive by exact number of ARAB (alcohol-resistant acid bacilli).To confirm the diagnosis of pulmonary tuberculosis, the molecular method GeneXpertMTB / RIF was used, finding a low load without resistance to Rifampicin. Through the analysis of this clinical case, we can reinforce the need to continue with the active search for M. tuberculosis even in acute pneumonias, since mixed infections are more common than is generally believed.

IDENTIFICATION GENE MUTATIONS RPOB CAUSE OF MULTIDRUG RESISTANCE TUBERCULOSIS IN HAJI ADAM MALIK HOSPITAL

Objective: The study aimed to investigate gene mutations in Mycobacterium tuberculosis causing resistance to rifampicin (RIF) in multidrug resistance TB patients in Haji Adam Malik Hospital and determined the suitability of the general pattern of mutations that occur in patients with multidrug-resistant TB (MDR-TB) in the hospital with global mutation pattern.Methods: This observational study was performed prospectively in 31 patients with MDR-TB who had data medical record results of GeneXpert MTB/ RIF positive for resistant to RIF period August–November 2016 in the hospital by taking sputum from patients. DNA extracted from sputum and then purified to be detected using polymerase chain reaction (PCR). PCR products were examined by electrophoresis.Results: The highest percentage of 81 bp rpoB gene mutation MTB is called as RIF -resistance determining region which was located at codon 516 (100%) and then was followed by mutation at codon 531 (96.77%) and codon 526 (90.32%), respectively, while the smallest percentage of mutation (12.90%) MTB at codon was 533.Conclusion: The rpoB gene shows a positive mutation-causing resistance to RIF.

Comparison of Gene Xpert MTB/RIF Assays with Conventional Standard Proportion Method for Determination of Drug Susceptibility In Multidrug Resistant TB Suspects

Background: Tuberculosis (TB) is a contagious disease and multidrug resistant tuberculosis is an emerging global issue. Rapid detection of such type of tuberculosis is necessary for timely control of the disease. GeneXpert test has already been implemented by World Health Organization to diagnose the infection on urgent basis. Objectives: This study was designed to apply GeneXpert MTB/RIF assays for the detection of rifampicin resistant tuberculosis and validation of assays by comparing with conventional standard drug proportion method. Additionally, to explore whether the assay can be utilized in treatment of Multidrug Resistant TB Settings: This study was undertaken in Pakistan Health Research Council TB Research Centre in collaboration with Department of Pulmonology, King Edward Medical University, Mayo Hospital, Lahore. Methods: Sputum samples from 125 patients were collected from confirmed pulmonary TB patients who were not responding to standard regimen of first line anti-tubercular treatment. Smears were stained by Ziehl-Neelsen method. All specimens were processed for culture and drug sensitivity by drug proportion method using Lowenstein Jensen medium as well as GeneXpert MTB/RIF assay. Results: A total of 125 subjects were registered in present study including 64 (51.2%) males and 61 (48.8%) females of age 15 years and above with mean age of 36.9±14.99. Sensitivity and specificity of the assay was observed as 92.1% and 93.5%, respectively. Association of rifampicin resistant by MTB/RIF assay and isoniazid resistance was found to be 88.1% and an agreement rate of rifampicin resistance by GeneXpert MTB/ RIF assay with isoniazid resistance was 81.25%. A total of 56 (44.8%) cases were found to be multidrug resistant patients and an agreement rate of 92.9% (52/56) was demonstrated in multidrug resistant patients which was found to be rifampicin resistant by GeneXpert in present study. Conclusion: GeneXpert MTB/RIF assay shown high sensitivity (96.7%) and specificity (98.6%). This most modern and latest technique, particularly in smear negative patients, helps rapid detection of TB and rifampicin resistance, which facilitates prompt diagnosis of multidrug resistant TB. These results propose that effective treatment can be initiated at an early stage, which will greatly help in reducing multidrug resistant TB.

Comparative Study between Genexpert and Smear Microscopy for the Diagnosis of Tuberculosis in Samples of Patients Suspected of Pulmonary Tuberculosis – the Case Study of General Hospital, Awo-Omamma, Imo State, Nigeria

Tuberculosis (TB), caused by Mycobacterium tuberculosis, has remained a major scourge of humanity all over the world, with the greatest mortality occurrences noted, in developing countries. The cannot-be-over- emphasized burden of TB in Nigeria is among the highest in Africa. The study on hand was therefore aimed at comparing Cepheid GeneXpert MTB/ RIF assay for direct detection of Mycobacterium tuberculosis Complex (MTBC) and Rifampicin (RIF) resistance with the traditional smear microscopy method-the ZN technique. Sensitivity and specificity of diagnostic yields were high points of comparison. A carefullydesigned cross-sectional study was drawn and executed at the General Hospital, Awo-Omamma, covering patients’ inflow from August, 2016 to May 2017. Amongst the numerous patients presenting, a total of 120 samples were collected from patients with highest pulmonary concerns, having been assessed prognostically. Sixty-two patients (51.67%) were males, fifty-eight (48.33%) were females and all having mean ages of 42.2+16 years. Thirty patients (25%) had chronic lung diseases. Out of the 120 samples examined, 36 samples (30.00%) were MTBC positive by Smear microscopy while 42 (35.00%) were positive by GeneXpert. Placing both methods (GeneXpert and Smear microscopy) side-by-side, GeneXpert gave 85% sensitivity and 98.5% specificity GeneXpert indeed detected 6 (7.2%) additional positive cases as compared to Smear microscopy. Only 5 clinical isolates of the entire patients were resistant to Rifampicin. The study therefore conduced that GeneXpert was a better and more reliable diagnostic tool compared to Smear microscopy and can significantly reduce false-negatives and very interestingly, rules out the unnecessary delays often experienced hitherto with Smear microscopy in treatment initiation.

Programmatic Impact of Implementing GeneXpert MTB/ RIF Assay for the Detection of Mycobacterium Tuberculosis in Respiratory Specimens from Pulmonary Tuberculosis Suspected Patients in Resource Limited Laboratory Settings of Eastern Nepal

Background:In Nepal, introduction of GeneXpert MTB/RIF assay (Xpert assay) as an initial confirmation test for tuberculosis (TB) has been considered to have impact as a significant decrease in number of clinically diagnosed pulmonary tuberculosis (PTB) cases than previous years. This study aims to find out the distribution profile of suspected tuberculosis cases according to patients age, gender, treatment history and HIV status as well as to evaluate the utility of the Xpert assay over conventional acid-fast bacilli (AFB) staining method for the proper diagnosis of M. Tuberculosis in respiratory specimens from the tuberculosis (TB) suspected patient samples. Methods:The prospective cross-sectional analytical study was conducted in National Anti-Tuberculosis Center (NATA) center- Biratnagar and Primary Healthcare Center (PHC) - Manglabare, Morang District, of eastern Nepal from January 2014 to August 2014. Laboratory investigation was done by conventional AFB staining followed by Xpert assay. Results:A total of 1549 sputum samples were initially analyzed. AFB staining resulted in 1441 AFB smear negative samples and 88 AFB smear positive samples, whereas 20 samples were directly processed for Xpert assay. The male: female smear positive ratio was 2.8:1 and was higher among age groups (21-40) years. Tuberculosis among HIV patients was found 22.22%. Xpert assay demonstrates that out of 1441 smear negative AFB cases, 258 were found to have TB positive, whereas out of 88 smears positive AFB cases 12 were found to have TB negative. The sensitivity of the Xpert assay in patients classified as AFB smear positive was found 85.4% and the specificity in smear negative patients was 81%. Conclusion:The study concluded that implementation of Gene Xpert MTB/RIF assay is a helpful tool for early and rapid detection of tuberculosis with greater sensitivity and specificity over traditional AFB staining techniques.

Comparative Study between Genexpert and Smear Microscopy for the Diagnosis of Tuberculosis in Samples of Patients Suspected of Pulmonary Tuberculosis – the Case Study of General Hospital, Awo-Omamma, Imo State, Nigeria

Tuberculosis (TB), caused by Mycobacterium tuberculosis, has remained a major scourge of humanity all over the world, with the greatest mortality occurrences noted, in developing countries. The cannot-be-over- emphasized burden of TB in Nigeria is among the highest in Africa. The study on hand was therefore aimed at comparing Cepheid GeneXpert MTB/ RIF assay for direct detection of Mycobacterium tuberculosis Complex (MTBC) and Rifampicin (RIF) resistance with the traditional smear microscopy method-the ZN technique. Sensitivity and specificity of diagnostic yields were high points of comparison. A carefullydesigned cross-sectional study was drawn and executed at the General Hospital, Awo-Omamma, covering patients’ inflow from August, 2016 to May 2017. Amongst the numerous patients presenting, a total of 120 samples were collected from patients with highest pulmonary concerns, having been assessed prognostically. Sixty-two patients (51.67%) were males, fifty-eight (48.33%) were females and all having mean ages of 42.2+16 years. Thirty patients (25%) had chronic lung diseases. Out of the 120 samples examined, 36 samples (30.00%) were MTBC positive by Smear microscopy while 42 (35.00%) were positive by GeneXpert. Placing both methods (GeneXpert and Smear microscopy) side-by-side, GeneXpert gave 85% sensitivity and 98.5% specificity GeneXpert indeed detected 6 (7.2%) additional positive cases as compared to Smear microscopy. Only 5 clinical isolates of the entire patients were resistant to Rifampicin. The study therefore conduced that GeneXpert was a better and more reliable diagnostic tool compared to Smear microscopy and can significantly reduce false-negatives and very interestingly, rules out the unnecessary delays often experienced hitherto with Smear microscopy in treatment initiation.