GeneXpert Research Articles

Diagnostic Value of Encode TB IgG and IgM Rapid Test to Support Pulmonary Tuberculosis Diagnosis

Diagnosis of tuberculosis can be established through the detection of antigens by Acid Fast Bacilli (AFB), microscopy,culture, and Polymerase Chain Reaction (PCR). The World Health Organization (WHO) 2012 issued a recommendation not touse antibody detection in the diagnosis of tuberculosis. However, there is high demand from clinicians to detectanti-tuberculosis antibody in patients who are challenging to do a bacteriological examination. The purpose of this researchwas to determine the diagnostic value of anti-M.tuberculosis IgG and IgM Encode TB to support lung tuberculosis diagnosis.This study was a cross-sectional by using consecutively sampling, which was performed in the Dr. Soetomo Hospital,Surabaya, Indonesia, from November 2017 until May 2018. A total of 52 patients were included and evaluated for clinical orbacteriological examination using AFB microscopy or PCR (Gene Xpert) as the gold standard and tested theanti-M.tuberculosis IgG and IgM with immunochromatography. Encode Tuberculosis (TB) IgG was positive in 12 patientsfrom the tuberculosis group and one false-positive in the non-tuberculosis group. The diagnostic sensitivity, specificity,positive predictive value, negative predictive value, and accuracy of Encode TB IgG dan IgM were 35%, 94%, 92%, 43% and55.7%, respectively. The specificity was high that the positive result was considered as TB; the sensitivity was low that thenegative results were not excluded from TB. Encode TB IgG/IgM rapid test was not recommended to use as a singlediagnostic test and must be combined with other diagnostic tests to increase the sensitivity.

Forty-Year-Old Man With Abdominal Pain 4 Years Post–Renal Transplant: A Case Report

Tuberculosis (TB) is an opportunistic infection 20 to 74 times more frequent in immunocompromised patients compared to the general population. The prevalence with renal transplant had a 0.5% to 15% incidence. The infection could be pulmonary or extrapulmonary (EPTB). The EPTB accounts for almost 20% of TB cases in immunocompetent people and 50% in positive human immunodeficiency virus cases. In this case report, we present a patient who attended the emergency room because of chronic diarrhea, abdominal pain, loss of weight, nocturne diaphoresis, and intermittent fever. A computed tomography scan showed retroperitoneal ganglionic conglomeration. He got into an exploratory laparotomy for histopathology specimens and paraganglionic fluid culture to a Gene Xpert MTB-RIF Assay G4, positive for rifampicin resistance tuberculosis. After an individualized treatment, trying to protect the graft’s remaining function, the patient returned with acute abdominal pain and pancreatic enzymes elevation; the antibiotic management had to be suspended until the return of renal function.

The Role of IL-6 Gene Polymorphism in Multidrug-Resistant Tuberculosis Patients in Iraq

Background: Tuberculosis (TB) remains to be a leading cause of morbidity and mortality in developing countries, and the incidence of the disease is increasing in developed. One-third of the world’s population is infected by Mycobacterium tuberculosis (Mtb). Still, only about 5% of infected individuals develop the disease within the first year of infection, and another 5% develop the disease later in life. Aim of the study: Is to investigate the relationship between the levels of Interleukin 6 (IL-6) and detection of multidrug-resistant tuberculosis (MDR-TB) in Al-Diwaniyah population. Patients and methods: The current study included 120 patients with tuberculosis who were classified into two groups. The first group included 60 TB patients who were sensitive to anti TB drugs and 60 patients with multi-drug resistance (MDR) based on gene Xpert. The study also included 60 healthy individuals serving as a control group. Demographic characteristics of study and control groups were retrieved. IL-6 (-572G/C) genotype polymorphism was carried out using polymerase chain reaction (PCR) and according to the instruction of the providing company. Serum level of IL-6 was also measured by ELISA according to the instruction of the providing company. Results: It has been observed that genotype CC was more frequent in sensitive TB patients than both MDR TB patients and control subjects, 73.3 % versus 60 % and 61.7 %, respectively. Serum IL-6 level was significantly higher in MDR TB patients than in control group (p = 0.007), and there was no significant difference in its level between sensitive and MDR TB patients (p = 0.284). Conclusion: IL-6 gene polymorphism is not associated with multidrug resistance in TB patients; however, Serum IL-6 level was significantly higher in MDR TB patients than in the control group.

A perspective of Mycobacterium Tuberculosis resistance on patients with retreat in Madagascar

Introduction: Despite advances in diagnostic and therapeutic way, drug-resistanttuberculosis is an increasingly worrying threat. Several studies suggest that one ofthe risk factors for carrying resistant Mycobacterium tuberculosis is the historyof anti-tuberculosis treatment. The objective of our study is to describe theResistance Profile of Mycobacterium Tuberculosis of patients in retreatment inMadagascar. Methods: This is a descriptive and retrospective study, carried outfrom January 2014 to September 2018 (45 months). All the cases of re-treatementrecorded in the database of the National Tuberculosis Control Program inMadagascar were included in the study.Results: We gathered 1456 cases, the average age was 40 years, there was a malepredominance(sex ratio of 2.32). In the 1302 cases where the category wasspecified, there was a predominance of relapse (76.57%). According to theculture, the incidence of rifampicin resistance was 05.36% (78/1456) including83.33% of cases of multi-resistant tuberculosis (MDR-TB) (65/78). The incidenceof isoniazid monoresistance was 05.49% (80/1456). Carrying MDR-TB is much moreimportant for patients who have failed (69.43%). The GeneXpert mtb/RIF enabledus to detect more cases of Rifampicine monoresistance (97 cases).Conclusion: cases of retreatment is a real source of the emergence of MDR-TB inMadagascar, especially cases of failure and relapse. Which is consistent with the data inthe literature. The treatment effectiveness of new cases must be maximum to preventthese cases of retreatment.Key words: drug resistance–Madagascar–profil–tuberculosis

Factors Contributing to Low Tuberculosis Case Detection in West Akim Municipality of the Eastern Region of Ghana: A Community and Health Facility Based Study

This research has a keen interest into assessing health and community related factors that contribute and influences low TB case detection rate in the West Akim Municipality of the Eastern region of Ghana. Data was collected from a target group of community members and health workers using a well-structured questionnaire. Data from the field was collected using Epi Info 7 and then, exported to Microsoft Excel and finally transferred to SPSS version 24 for the analysis. Some statistical techniques such as descriptive statistics, reliability of scale, t-test of independence, chi-square and analysis of variance was used to obtain results from the data analysis. Community related factors that has been identified as possible contributors to low TB detection rate are as follows, the availability of alternative means of health care (traditional healers, prayer camps, and herbs to manage suspected cases of TB. This turns to keep people away from the health facilities. The long time (2 days) it takes for an individual to get tested and to receive his/her tested results (time of diagnostics). The high levels of experienced stigma associated with psychological distress on suspected people with TB and on friends and relatives living with TB patients. the low knowledge of TB among some reasonable number of community members. The study also identified the possible health related factors influencing low TB case detection rate such as, Low TB trained personals and low TB education during health talks in creating awareness that TB is preventable, curable and treatable and that suspected people with TB (cough for more than two weeks) should seek medical attention. Community health nurses not including TB screening as part of their home visitation program and low level of contact tracing could also contribute to the low case detection rate. The highly inadequate number Gene Xpert machines, DOT centres and laboratories for the diagnostics of TB. The study also found that, TB stigma even existed among health care providers against suspected patients. This negatively influence the health seeking behavior among suspected patients. This study became necessary to generate information for the Ghana national TB control Programme for appropriate remedial actions to be taken to forestall any catastrophe that may result from undetected and untreated TB cases in the West Akim municipality. Keywords: Gene Xpert machines, DOT Centres, Stigmatization, Psychological Distress, TB screening, Low TB Case Detection. DOI: 10.7176/JHMN/72-11 Publication date: March 31 st 2020

Comparison Study of GENEXPERT versus TB MGIT Culture in Extra Pulmonary Tuberculosis

Aims and objectives: To compare the diagnostic yield of Extra Pulmonary Tuberculosis, in terms of MTB isolation & detection of drug resistance via GENEXPERT & MGIT-DST culture methods. Materials and Methods: This was a prospective observational study carried out in Department of Pulmonary medicine, D.Y. Patil Hospital, Navi Mumbai. Study participants were patients above 18 years of age who had extra pulmonary tuberculosis and who were not already on Anti tuberculosis treatment in D.Y. Patil Hospital, Nerul, Navi Mumbai and willing to participate in the study and meeting all the inclusion criteria of the study. Study duration was from the date of approval by institutional ethics committee to October 2019.Sample size was 150 patients satisfying the inclusion criteria. Results: In our study, 28 % of all extra pulmonary samples were tested positive for TB MGIT culture out of which 35.2 % showed Resistance to 1st line ATT drugs on DST where as 39.33% was tested positive with Gene expert out of which 22.47% showed Rifampicin Resistance. Sensitivity of MGIT was 28.00 % and specificity was 39.50 %.Sensitivity of Genexpert was 39.33% and specificity was 26.5%. Conclusion: Our findings suggest that Gene Xpert may have a role in EPTB diagnosis in addition to PTB, particularly in low income/high-burden settings, where facilities for mycobacterial culture are limited. But Gene Xpert can detect only Rifampicin resistance where as DST by BACTEC MGIT AFB Culture detects other ATT drugs sensitivity too. Also the test results of MTB detected in GeneXpert in extra pulmonary samples was shown as low and very low based on the CT range (high, 28) as most of the extra pulmonary samples are pauci bacillary; hence the result is not totally reliable. Therefore, it should be confirmed by phenotypic DST by BACTEC MGIT Culture.

Diagnostic Accuracy of Gene Xpert vs Acid Fast Smear in HIV Patients Suspected for Tuberculosis

Diagnostic Accuracy of Gene Xpert vs Acid Fast Smear in HIV Patients Suspected for Tuberculosis

The use of GeneXpert Mycobacterium tuberculosis/rifampicin assay in diagnosis of pulmonary tuberculosis in children

Introduction Rapid diagno sis of tuberculosis (TB) is essential for early disease management. GeneXpert (GX) test is a novel rapid diagnostic modality. In this study, we aimed to evaluate the diagnostic role of GX test in diagnosing pulmonary tuberculosis (PTB) in children as well as detection of rifampicin (RIF) resistance. Patients and methods This was a cross-sectional study that included 80 patients suspected to have PTB based on clinical data together with at least one of the followings: a positive tuberculin skin test or QuantiFERON-TB Gold In-Tube test result, a positive blood TB PCR result, or a positive Ziehl–Neelsen staining result. Results PTB was more common in older females. Extrapulmonary manifestations and hilar lymphadenopathy were more frequent in younger age. Both positive tuberculin skin test and positive GX were more frequent in older age. Positive GX was found in 30 (37.5%) cases, and RIF resistance was detected in two (6.6%) of them. Positive GX results were more frequent in sputum than gastric aspirate samples. Positive GX results were more frequent with absent Bacillus Calmette–Guerin scar, and with hemoptysis, failure to thrive, cavitary lung lesions, and lower mean hemoglobin. Conclusion GX detects many more cases than blood PCR and Ziehl–Neelsen stain. GX had 70% sensitivity and 95% specificity in diagnosing PTB, with overall diagnostic accuracy of 86.67%, with better performance in older children who could expectorate. RIF resistance was ∼6.6%.

A Study on Diagnostic Evaluation of Two Different Rapid DNA Polymerase Chain Reaction Techniques Namely Gene Xpert Mycobacterium Tuberculosis/Rifampin (MTB/RIF) and Mycoreal Polymerase Chain Reaction in the Diagnosis of Endometrial Tuberculosis Considering Culture as Gold Standard.

Context:The study involves the evaluation of two polymerase chain reaction (PCR) techniques one of which has been endorsed by the WHO for their diagnostic capabilities.Aims:The aim of this study is to evaluate the diagnostic accuracy of GeneXpert mycobacterium tuberculosis/Rifampin (MTB/RIF) and mycoreal PCR techniques in the diagnosis of endometrial tuberculosis (TB) considering culture as the gold standard.Settings and Design:A retrospective study conducted at Gunasheela surgical and maternity hospital. Patients who attended the outpatient department between January 2013 and August 2016, satisfying the eligibility criteria, were included in the study.Methodology:Women included in the study underwent endometrial pipelle sampling premenstrually after ruling out pregnancy in that cycle. Endometrial samples were tested for TB by Mycoreal PCR, Gene Xpert and BACTEC culture.Statistical Analysis Used:Statistical analysis was done using the R software version 3.6.1. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of test were calculated.Results:A total of 3229 samples were analyzed, of which 1754 were evaluated by Mycoreal TB PCR and 1475 were evaluated by Gene Xpert MTB/RIF assay. The sensitivity of mycoreal TB PCR technique was 34.78%, specificity was 99.08%, PPV was 33.33%, NPV was 99.13%, and accuracy was 98.23%. The sensitivity of GeneXpert MTB/RIF technique was 6.90%, specificity was 99.79%, PPV was 40.00%, NPV was 98.16%, and accuracy was 97.97%.Conclusions:MYCOREAL seemed to be more sensitive than Gene Xpert (MTB/RIF) considering culture as the gold standard in the diagnosis of endometrial TB.

Role of Gene Xpert in the Detection of Genital Tuberculosis in Endometrial Tissue among Women with Infertility.

Objective:The objective of the study is to evaluate the clinical utility of Gene Xpert compared with other available diagnostic modalities in prompt diagnosis of female genital tuberculosis (TB) causing infertility.Material and Methods:This was a prospective, cross-sectional analytical study. Premenstrual endometrial biopsy specimens were collected from 176 infertile women of reproductive age group suspected of having genital TB. Samples were processed for acid-fast bacilli, culture, histopathology, polymerase chain reaction (PCR), and Gene Xpert. Patients detected positive on Gene Xpert and PCR were subjected to laparoscopy to look for affirmative findings of genital TB. The results were analyzed using composite gold standard consisting of patients positive with culture, histopathology, and laparoscopy.Results:A total of 18 patients were found positive using composite gold standard. Laparoscopy was positive in 15 patients, whereas culture and histopathology were positive in three and two patients, respectively. Gene Xpert was positive in two patients. None of them was detected with rifampicin resistance. The sensitivity of Gene Xpert was 11.11% whereas the specificity was 100%.Conclusion:Since genital TB is a paucibacillary disease, multiple diagnostic modalities are needed for diagnosis. Gene Xpert appears to be a useful modality in diagnosis of genital TB, owing to its high specificity, and can be recommended in conditions where microscopy, culture, and histopathology are negative; however, further randomized studies are required to support our hypothesis.

ASSESSMENT OF THE MICROSCOPIC– OBSERVATION- DRUG - SUSCEPTIBLITY ASSAY (MODS) AND GENEXPERT ASSAY FOR DIAGNOSIS OF PULMONARY TUBERCULOSIS

Background: Tuberculosis (TB) is considered the most common and highly contagious disease worldwide. Early and accurate diagnosis of TB with detection of multidrug-resistant Mycobacterium tuberculosis (M.TB) is of primary importance for both patient management and infection control. Objectives: To evaluate the performance of Microscopic-Observation Drug-Susceptibility (MODS) assay and GeneXpert MTB/RIF( GeneXpert ) assay in diagnosis of TB and detection of MDR-TB directly from sputum specimens in comparison to the convential diagnostic methods (direct Ziehl-Neelsen "Z-N" stained smear and culture on Lowenstein-Jenson media "LJ"). Patients and Methods: After approval of institutional ethical committee a total of 120 suspected pulmonary TB patients admitted to chest hospitals and dispensaries in Egypt (Giza, Cairo, Alexandria, Dakahlia, Suez, Beheira, Monufia, Qena). Their age was more than 16 years; all of them did not receive any anti tubercular drugs. Sputum samples were collected from each patient and subjected to direct ZN smear, culture and sensitivity of M.TB on LJ and Middlebrook 7H9 medium for MODS assay. Finally Genexpert M.TB/RIF assay was done directly for all sputum samples. Results: Infection rate among males was higher than female (81.7% & 18.7%). Also, it was observed that 56.7% of sputum samples gave positive direct Z-N smear, while 64.2% were positive for both LJ culture &MODS assay. On the other hand, by Genexpert, 68% of cases were positive. The sensitivity and specificity of the various diagnostic methods were studied in relation to LJ culture (gold standard). It was found that MODS assay was rapid that results were obtained in a median of 8 days ranged from (7– 21 days), while median time of LJ was 21dayes ranged from (21– 60). MODS had 100% sensitivity and specificity. Direct smear was less sensitive (88.3%), and GeneXpert was less specific (88.4%), but more rapid results were obtained in 2 hours. Finally, detection of MDR-TB using various methods for 77 isolated M.TB strains ranged from 16.9-19.5% to isoniazid (INH) and 9.8%-18.2% to rifampicin (RMP) by various diagnostic methods with statistically insignificant difference between them. Conclusion: MODS was an optimal alternative rapid diagnostic technique for TB and detection of M.TB and MDR-TB in resource limited settings, results were obtained in a median of 8 days. On the other hand, Gene Xpert was a novel integrated diagnostic PCR analysis in a single hand free step for rapid diagnosis of TB and detection of RMP resistance in clinical specimens in 2 hours but it was expensive.

ASSESSMENT OF THE MICROSCOPIC– OBSERVATION- DRUG - SUSCEPTIBLITY ASSAY (MODS) AND GENEXPERT ASSAY FOR DIAGNOSIS OF PULMONARY TUBERCULOSIS

Background: Tuberculosis (TB) is considered the most common and highly contagious disease worldwide. Early and accurate diagnosis of TB with detection of multidrug-resistant Mycobacterium tuberculosis (M.TB) is of primary importance for both patient management and infection control. Objectives: To evaluate the performance of Microscopic-Observation Drug-Susceptibility (MODS) assay and GeneXpert MTB/RIF( GeneXpert ) assay in diagnosis of TB and detection of MDR-TB directly from sputum specimens in comparison to the convential diagnostic methods (direct Ziehl-Neelsen stained smear and culture on Lowenstein-Jenson media LJ). Patients and Methods: After approval of institutional ethical committee a total of 120 suspected pulmonary TB patients admitted to chest hospitals and dispensaries in Egypt (Giza, Cairo, Alexandria, Dakahlia, Suez, Beheira, Monufia, Qena). Their age was more than 16 years; all of them did not receive any anti tubercular drugs. Sputum samples were collected from each patient and subjected to direct ZN smear, culture and sensitivity of M.TB on LJ and Middlebrook 7H9 medium for MODS assay. Finally Genexpert M.TB/RIF assay was done directly for all sputum samples. Results: Infection rate among males was higher than female (81.7% & 18.7%). Also, it was observed that 56.7% of sputum samples gave positive direct Z-N smear, while 64.2% were positive for both LJ culture &MODS assay. On the other hand, by Genexpert, 68% of cases were positive. The sensitivity and specificity of the various diagnostic methods were studied in relation to LJ culture (gold standard). It was found that MODS assay was rapid that results were obtained in a median of 8 days ranged from (7– 21 days), while median time of LJ was 21dayes ranged from (21– 60). MODS had 100% sensitivity and specificity. Direct smear was less sensitive (88.3%), and GeneXpert was less specific (88.4%), but more rapid results were obtained in 2 hours. Finally, detection of MDR-TB using various methods for 77 isolated M.TB strains ranged from 16.9-19.5% to isoniazid (INH) and 9.8%-18.2% to rifampicin (RMP) by various diagnostic methods with statistically insignificant difference between them. Conclusion: MODS was an optimal alternative rapid diagnostic technique for TB and detection of M.TB and MDR-TB in resource limited settings, results were obtained in a median of 8 days. On the other hand, Gene Xpert was a novel integrated diagnostic PCR analysis in a single hand free step for rapid diagnosis of TB and detection of RMP resistance in clinical specimens in 2 hours but it was expensive.

18F-FDG PET/CT as a modality for the evaluation of persisting raised infective markers in patients with spinal tuberculosis

Aims: The aim of the study was to investigate the differences in participant characteristics between positive and negative, positron emission tomography with 2-deoxy-2-[fluorine-18]fluoro-D-glucose integrated with computed tomography (18F-FDG PET/CT) activity at the spinal tuberculosis (TB) site following 12 months of the appropriate chemotherapy therapy for spinal TB. A secondary aim of the study was to determine whether erythrocyte sedimentation rate (ESR) levels could be used as a reliable marker of TB activity and/or treatment success of spinal TB, especially in a high HIV-positive population. Patients and methods: All patients who were treated for spinal TB and underwent an 18F-FDG PET/CT scan were considered for inclusion. PET/CT positive patients underwent a spinal biopsy which was sent for microscopy, Gram staining, Gene Xpert (GXP) polymerase chain reaction (PCR) and histology. Patients in the PET/CT positive group underwent a repeat MRI scan and biopsy at the completion of treatment to investigate the potential presence of resistance or ongoing active spinal TB. Results: A total of 18 patients were included in the study: five patients were allocated to the PET/CT positive group and 13 to the PET/CT negative group. The PET/CT negative group was significantly older (p=0.016) and had significantly fewer TB-infected vertebrae (p=0.010) than the PET/CT positive group. Two patients, one in each group, were found to have drug-resistant spinal TB. At the 12-month follow-up visit, two patients (40%) in the PET/CT positive group and three patients (30%) in the PET/CT negative group were still complaining of back pain. All smear microscopy results of the PET/CT positive patients who underwent a repeat biopsy were negative after the conclusion of treatment; culture results (n=4/4) were also negative. GXP PCR results were positive in four and negative in one case. Only one of four samples showed classic TB signs on histology. Conclusion: This study is the first to report on biopsies done from a PET/CT positive site, after 12 months of anti-tubercular treatment. It is not unlikely that PET/CT is over-sensitive and can show metabolic activity in areas of sterile inflammation, and future studies are necessary to evaluate this. Level of evidence: Level 3

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.

Cartridge-based nucleic acid amplification test (Xpert Mycobacterium tuberculosis/Rifampicin Assay): An essential molecular diagnostic test for early diagnosis and initiation of treatment in childhood tuberculous meningitis and primary multidrug-resistant cases

Background: Tuberculosis (TB) still remains the major public health threat in India. Early diagnosis, so as to initiate early treatment is a priority as any delay, may complicate the prognosis further leading to the failure of an effective control. India accounts for 6% incidence of pediatric TB cases in a population that has 40% as estimated latent TB cases. Pediatric samples were considered as the key population in the study. TB remains the most common cause of childhood meningitis in high burden countries. India accounted for the highest number of TB deaths among children in the year 2015, followed by Nigeria and China. Of which, tuberculous meningitis (TBM) stands to be the most common cause of morbidity. Extrapulmonary TB accounts for 25% of the total TB cases reported among which the most severe form being TBM. Although severe and difficult to diagnose due to lack of specific tests to get confirmed, cartridge-based nucleic acid amplification test (CBNAAT) is the only solution for microbiological confirmation of the disease for early treatment initiation and reduction of morbidity and mortality. Previously, due to lower case detection rates owing to inadequate diagnostic facilities of higher analytical sensitivity, the mortality and morbidity continued to be in the higher range, and empirical treatment modality was the mainstay till the CBNAATs came into existence. The increase in the case detection for tuberculous meningitis at a manageable stage of the disease would help in decreasing the mortality rates. The study explores the increased case detection rates among pediatric population suffering from tuberculous meningitis from referral units to Intermediate Reference Laboratory (IRL), Kolkata. Cerebrospinal fluid (CSF) from the pediatric patients has not been very difficult procedure though invasive, especially in the pediatric age group, and can be practiced at peripheral unit having CBNAAT facility. Methods: CSF from pediatric patients showing obvious clinical signs of meningitis was collected from tertiary unit linked to IRL, Kolkata, and tested on CBNAAT by Xpert Mycobacterium tuberculosis/ rifampicin (RIF) (Cepheid) along with all the pulmonary and extrapulmonary samples based on the manufacturer's instruction. The data were captured in the Inbuilt software of the GeneXpert MTB/RIF (Xpert) and Genotype MTBDRplus (DRplus) (Cepheid , USA) of the cartridge based nucleic acid amplification test machines automatically during the tests and exported to the Microsoft Excel sheets for further analysis. A defined study design against each and every objective was setup to analyze the effective detection of tuberculous meningitis case and their resistance to RIF along with the other pulmonary and extrapulmonary samples. Results and Conclusions: The study concluded the confirmation of tuberculous meningitis cases by the help of CBNAAT. The detection of microbiologically confirmed TB by these specimens sent from the referral unit linked with IRL, Kolkata, for pulmonary and extrapulmonary cases along with tuberculous meningitis cases is significant and facilitates the early initiation of treatment for TB and more so to those who are newly detected as multidrug-resistant TB especially below the 6 months of age. About 5% prevalence and around 4.9% incidence are an alarming situation in the TB control scenario of West Bengal. The positive case detection among the CSF samples was found to be more in the 0.6–1-year group followed by the age group of 1–5 years. The percentage positivity of CSF is recorded as 2.38%, and the percentage of RIF resistance among positive cases from CSF was found to be 13.79% (n = 28).

The Outcome and Hepatic Function of an Extended Category II Treatment in a HIV-Seronegative Suspected Extensively Drug-Resistant TB Patient

A 25 year old HIV- seronegative client was diagnosed with pulmonary tuberculosis and placed on Category I (CAT I) treatment. After two months, acid fast bacilli (AFB) persisted in sputum. Rifampicin (Rif) resistance was detected with the GeneXpert MTB-Rif Assay® (Xpert; Cepheid, Sunnyvale, CA, USA), leading to a change in treatment to CAT II after completing CAT I in the third month. Continuous detection of Rif resistant species and persisting ‘Scanty’ results (1-9 AFB found in 100 microscopic fields) from sputum smear microscopy (SSM) after one (1) year was suggestive of treatment failure to CAT II [possible extensively drug-resistant (XDR) species]. Category II treatment was continued for the next eight months as we anticipated the arrival of XDR medicines. Before commencement of XDR medication, SSM and GeneXpert were repeated. The SSM result was negative (no AFB seen), while Rif resistance persisted. The controversy was eventually clarified with the Bactec Mycobacteria Growth Indicator Tube (MGIT) liquid culture, which indicated no growth for M. tuberculosis. Since the rapid molecular GeneXpert will persistently detect Rif resistance even after months of treatment, we support the use of MGIT to confirm when treatment should be halted. This case study also showed tolerance to prolonged treatment with CAT I and II drugs and confirms evidence in existing literature.

Pattern of Childhood Tuberculosis in the Outpatient Department of a Tertiary Level Hospital in Dhaka City

Introduction: Tuberculosis (TB) in children is increasingly becoming an important cause of global child morbidity and mortality. Objective: The objective of this study was to evaluate the clinical spectrum of TB in children under the age of 15 years and document any changes that occur over time. Materials & Methods: This observational study was conducted in the pediatric outpatient department (OPD) of 250 Bedded TB Hospital, Shyamoli, Dhaka, from October’2016 to January’2017. A total 71 children of both sex up to 15 years of age, who were diagnosed as having TB and attended the pediatric OPD of TB hospital during the four months study period were enrolled. The data was analyzed on the basis of patient’s age, gender, socioeconomic status, mode of presentation, BCG vaccination status, history of contact with smear positive TB patient, clinical findings, investigations and associated co-morbidities. Results: This study revealed that among the 71 cases of TB, (72%) had extra-pulmonary TB (EPTB) and (28%) had pulmonary TB (PTB), the commonest age group was 6 to 15 years with male preponderance (51%). Pulmonary TB was diagnosed mostly clinically (60%) followed by positive sputum smear result (20%), gastric aspirate for acid-fast bacilli AFB (5%) and sputum for Gene Xpert (5%). Distribution of extra-pulmonary TB (EPTB) according to the organ involvement was TB lymphadenitis (49%), osteoarticular TB (19.6%) and abdominal TB (5.9%). Cervical lymphadenopathy was the commonest presentation (76%) among the EPTB cases. Tuberculin skin test (TST) was positive in (69%) cases. Among the total cases (90%) children had TB alone whereas (10%) had other co-morbid disease along with TB. Conclusion: Extra-pulmonary TB (EPTB) was more prevalent among the childhood TB cases in a tertiary level set up where cervical TB lymphadenitis was the commonest.

HPV-Positive Laryngeal Carcinomas: Epidemiological, Virological and Progressive Features

Objective: The aim of this work was to determine the molecular prevalence of the Human Papilloma Virus (HPV) in carcinomas of the larynx and to identify its circulating genotypes. Patients and Methods: This was a descriptive, retrospective study of 20 years which included all the patients followed in otolaryngology department for laryngeal carcinoma. The virological analysis was done on the laryngeal biopsy pieces included in paraffin using the Gene Xpert technique. This technique uses real-time PCR to identify oncogenic HPV genotypes. Results: A total of 108 patients with laryngeal carcinomas were collected. Among them, 21 samples were associated with oncogenic HPV, a molecular prevalence of 19.4%. These were the HPV 18/45 (14.28%), HPV-16 (28.57%) and the group of HPVs with high oncogenic risk other than 18/45 and 16 (57.15%). All of these patients were male, mean age was 35.71 ± 3.17, and 85.7% of them were under 40 compared to patients with carcinomas not associated with HPV (p = 0.0003). Oral-genital contact was the main risk factor for contamination in all of these patients (p = 0.0002). The HPV-positive laryngeal carcinomas were all micro-invasive and the patients had better survival compared to those who had HPV-negative carcinomas (p = 0.0001). Conclusion: HPV-positive laryngeal carcinomas are most often observed in subjects under 40 years of age with good survival at 12 months. The circulating genotypes in Brazzaville are 16, 18/45 and the group of oncogenic HPVs other than 16 and 18/45.

Vitamin D level and it is association with the severity of pulmonary tuberculosis in patients attended to Kosti Teaching Hospital, Sudan.

Globally, tuberculosis is one of the major causes of morbidity and mortality in many countries. Previous studies suggest that the incidence and severity of tuberculosis are associated with low levels of vitamin D (Vit D). Therefore, this study aimed to determine the occurrence and associated factors of vitamin D3 deficiency in pulmonary tuberculosis patients at White Nile State, Sudan. 101 individuals of diagnosed pulmonary tuberculosis patients (71 males and 30 females) and 100 non-TB controls (58 males and 42 females) were included in this study. Sputum samples were obtained from TB patients and subjected to examination for acid-fast bacilli (AFB) using Ziehl-Neelsen (ZN) stain and Gene Xpert analysis. Blood samples were collected from both groups and Serum 25(OH)-vitamin D3 was determined by an Enzyme-Linked Immunosorbent Assay. HIV infection in Tuberculosis (TB) group was also investigated using the immunochromatographic test. In our study, the majority of TB patients were suffered from TB relapse (36.6%); non-HIV infected individuals (99.1%) or showed a positive result for AFB (61.4%) in Gene Xpert analysis. Moreover, there is a significant difference in microscopy findings and bacillary levels of AFB, and Rifampicin (RIF) susceptibility pattern of M. tuberculosis strain among sputum samples of TB patients, P-values less 0.0001. Furthermore, we found that TB patients were suffered from vitamin D deficiency. In particular, the mean of vitamin D level was significantly much lower in TB patients (26.7 ± 1.6) compared to non-TB controls (117.3 ± 3.2), P-value equal 0.0001. Likewise, it's much lower in females, individuals of 21–40 years old, and patients with high bacillary levels or those infected by Rifampicin resistance strain. Accordingly, our study was highlighted the TB and Vit D deficiency relationship and showed the need for further studies to a better understanding of the impact of TB on Vit D level and investigate whether vitamin D supplementation can have a role in the prevention and treatment of tuberculosis.

Isolated Hepatosplenic Tuberculosis: A Rare Case Report

Background: Tuberculosis is common in Bangladesh. It rarely involves the liver and spleen and when it does so, it is usually associated with disseminated disease. Method: We report a patient with isolated tuberculosis of the liver and spleen. Results: A 55 years old female presented with fever with chill and rigor and upper abdominal pain. USG revealed multiple hypoechoic lesions with central echogenicity in liver and spleen. CT scan showed multiple hypo density lesions in the liver and spleen. AFP, CEA and CA-19-9 were normal. As temperature was not subsided with antibiotics, pus from liver and spleen was aspirated and sent for gram and AFB stain, C/S, malignant cell, and Gene Xpert. All but Gene Xpert reports were negative. A detailed workup failed to identify other focus of tuberculosis. CAT-1 Anti tubercular started and the fever subsided. The patient gained weight on the subsequent follow up. Conclusion: Tuberculosis should also be considered in the differential diagnosis for multiple nodules in the liver and spleen along with other diagnosis, though it is rare. Gene X pert is a good diagnostic tool.