Analysis of the application of a gene chip method for detecting Mycobacterium tuberculosis drug resistance in clinical specimens: a retrospective study

AbstractMost Mycobacterium tuberculosis (Mtb) resistant to rifampicin (RIF) has mutations in the rpoB gene, while most Mtb resistant to isoniazid (INH) has mutations in the katG gene or inhA promoter. We used gene chip technology to detect mutations in these genes to determine the resistance of Mtb to RIF and INH. A total of 4148 clinical specimens with sputum smear positivity for acid-fast bacilli (AFB) were detected. Then, taking the results of the drug sensitivity test (DST) as the reference standard, the detection efficiency of sputum samples from different grades of positive smears was compared in detail. We found that the sensitivity of the gene chip method for detecting sputum samples with a grade ≥ AFB 2 + was higher than that of sputum samples with a grade ≤ AFB 1 + (P < 0.05). When the grade of the sample was ≤ AFB 1 +, the sensitivity of the gene chip method was 72.6% for RIF, 67.3% for INH, and 60.0% for MDR-TB. When the grade of the sample was ≥ AFB 2 +, the sensitivity of the gene chip method was 84.5% for RIF, 78.2% for INH, and 73.9% for MDR-TB. The results show that gene chip technology can be directly used to diagnose drug-resistant tuberculosis in clinical specimens, and the diagnostic efficiency for the detection of sputum specimens with a grade ≥ AFB 2 + is better than that of other sputum specimens.

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Multicenter Study of the Accuracy of the BD MAX Multidrug-resistant Tuberculosis Assay for Detection of Mycobacterium tuberculosis Complex and Mutations Associated With Resistance to Rifampin and Isoniazid

Clinical Infectious Diseases ◽

10.1093/cid/ciz932 ◽

2019 ◽

Vol 71 (5) ◽

pp. 1161-1167 ◽

Cited By ~ 5

Author(s):

Maunank Shah ◽

Sonia Paradis ◽

Joshua Betz ◽

Natalie Beylis ◽

Renu Bharadwaj ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Sensitivity And Specificity ◽

High Sensitivity ◽

Drug Susceptibility ◽

Multidrug Resistant ◽

Drug Susceptibility Testing ◽

Sputum Smear ◽

Smear Microscopy ◽

Resistant Tuberculosis ◽

Mdr Tb

Abstract Background Tuberculosis (TB) control is hindered by absence of rapid tests to identify Mycobacterium tuberculosis (MTB) and detect isoniazid (INH) and rifampin (RIF) resistance. We evaluated the accuracy of the BD MAX multidrug-resistant (MDR)-TB assay (BD MAX) in South Africa, Uganda, India, and Peru. Methods Outpatient adults with signs/symptoms of pulmonary TB were prospectively enrolled. Sputum smear microscopy and BD MAX were performed on a single raw sputum, which was then processed for culture and phenotypic drug susceptibility testing (DST), BD MAX, and Xpert MTB/RIF (Xpert). Results 1053 participants with presumptive TB were enrolled (47% female; 32% with human immunodeficiency virus). In patients with confirmed TB, BD MAX sensitivity was 93% (262/282 [95% CI, 89–95%]); specificity was 97% (593/610 [96–98%]) among participants with negative cultures on raw sputa. BD MAX sensitivity was 100% (175/175 [98–100%]) for smear-positive samples (fluorescence microscopy), and 81% (87/107 [73–88%]) in smear-negative samples. Among participants with both BD MAX and Xpert, sensitivity was 91% (249/274 [87–94%]) for BD MAX and 90% (246/274 [86–93%]) for Xpert on processed sputa. Sensitivity and specificity for RIF resistance compared with phenotypic DST were 90% (9/10 [60–98%]) and 95% (211/222 [91–97%]), respectively. Sensitivity and specificity for detection of INH resistance were 82% (22/27 [63–92%]) and 100% (205/205 [98–100%]), respectively. Conclusions The BD MAX MDR-TB assay had high sensitivity and specificity for detection of MTB and RIF and INH drug resistance and may be an important tool for rapid detection of TB and MDR-TB globally.

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Outcomes of Community-Based Systematic Screening of Household Contacts of Patients with Multidrug-Resistant Tuberculosis in Myanmar

Tropical Medicine and Infectious Disease ◽

10.3390/tropicalmed5010002 ◽

2019 ◽

Vol 5 (1) ◽

pp. 2

Author(s):

Nang Thu Thu Kyaw ◽

Aung Sithu ◽

Srinath Satyanarayana ◽

Ajay M. V. Kumar ◽

Saw Thein ◽

...

Keyword(s):

Chest Radiography ◽

Multidrug Resistant ◽

Sputum Smear ◽

Community Based ◽

Household Contacts ◽

Resistant Tuberculosis ◽

Symptom Screening ◽

Multidrug Resistant Tuberculosis ◽

Screening Algorithm ◽

Mdr Tb

Screening of household contacts of patients with multidrug-resistant tuberculosis (MDR-TB) is a crucial active TB case-finding intervention. Before 2016, this intervention had not been implemented in Myanmar, a country with a high MDR-TB burden. In 2016, a community-based screening of household contacts of MDR-TB patients using a systematic TB-screening algorithm (symptom screening and chest radiography followed by sputum smear microscopy and Xpert-MTB/RIF assays) was implemented in 33 townships in Myanmar. We assessed the implementation of this intervention, how well the screening algorithm was followed, and the yield of active TB. Data collected between April 2016 and March 2017 were analyzed using logistic and log-binomial regression. Of 620 household contacts of 210 MDR-TB patients enrolled for screening, 620 (100%) underwent TB symptom screening and 505 (81%) underwent chest radiography. Of 240 (39%) symptomatic household contacts, 71 (30%) were not further screened according to the algorithm. Children aged <15 years were less likely to follow the algorithm. Twenty-four contacts were diagnosed with active TB, including two rifampicin- resistant cases (yield of active TB = 3.9%, 95% CI: 2.3%–6.5%). The highest yield was found among children aged <5 years (10.0%, 95% CI: 3.6%–24.7%). Household contact screening should be strengthened, continued, and scaled up for all MDR-TB patients in Myanmar.

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Phase I, Single-Dose, Dose-Escalating Study of Inhaled Dry Powder Capreomycin: a New Approach to Therapy of Drug-Resistant Tuberculosis

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02346-12 ◽

2013 ◽

Vol 57 (6) ◽

pp. 2613-2619 ◽

Cited By ~ 78

Author(s):

Ashwin S. Dharmadhikari ◽

Mohan Kabadi ◽

Bob Gerety ◽

Anthony J. Hickey ◽

P. Bernard Fourie ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Single Dose ◽

Phase I ◽

Plasma Concentrations ◽

Dry Powder ◽

Content Type ◽

Resistant Tuberculosis ◽

Drug Concentrations ◽

Mdr Tb ◽

Dose Escalating

ABSTRACTMultidrug-resistant tuberculosis (MDR-TB) threatens global TB control. The lengthy treatment includes one of the injectable drugs kanamycin, amikacin, and capreomycin, usually for the first 6 months. These drugs have potentially serious toxicities, and when given as intramuscular injections, dosing can be painful. Advances in particulate drug delivery have led to the formulation of capreomycin as the first antituberculosis drug available as a microparticle dry powder for inhalation and clinical study. Delivery by aerosol may result in successful treatment with lower doses. Here we report a phase I, single-dose, dose-escalating study aimed at demonstrating safety and tolerability in healthy subjects and measuring pharmacokinetic (PK) parameters. Twenty healthy adults (n= 5 per group) were recruited to self-administer a single dose of inhaled dry powder capreomycin (25-mg, 75-mg, 150-mg, or 300-mg nominal dose) using a simple, handheld delivery device. Inhalations were well tolerated by all subjects. The most common adverse event was mild to moderate transient cough, in five subjects. There were no changes in lung function, audiometry, or laboratory parameters. Capreomycin was rapidly absorbed after inhalation. Systemic concentrations were detected in each dose group within 20 min. Peak and mean plasma concentrations of capreomycin were dose proportional. Serum concentrations exceeded 2 μg/ml (MIC forMycobacterium tuberculosis) following the highest dose; the half-life (t1/2) was 4.8 ± 1.0 h. A novel inhaled microparticle dry powder formulation of capreomycin was well tolerated. A single 300-mg dose rapidly achieved serum drug concentrations above the MIC forMycobacterium tuberculosis, suggesting the potential of inhaled therapy as part of an MDR-TB treatment regimen.

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Association of Socio-Demographic Profile with Prevalence of Multi Drug Resistant Tuberculosis among Retreated Pulmonary Tuberculosis Patients in North India

SAARC Journal of Tuberculosis Lung Diseases and HIV/AIDS ◽

10.3126/saarctb.v16i1.23238 ◽

2018 ◽

Vol 16 (1) ◽

pp. 1-5

Author(s):

A. Chaudhary ◽

T. Mahmood ◽

A.D. Shukla ◽

A. Shreenivasa ◽

Verma Arvind ◽

...

Keyword(s):

Cross Sectional Study ◽

North India ◽

Sputum Smear ◽

Cross Sectional ◽

Pulmonary Tb ◽

Demographic Profile ◽

Resistant Tuberculosis ◽

Multi Drug Resistant Tuberculosis ◽

Mdr Tb ◽

Education Status

Methods: An observational cross sectional study, which includes 116 patients of sputum smear positive pulmonary TB of age 18 or above. Further, detailed history taking regarding different demographic profile was done. Also, they were subjected to CB-NAAT and rifampicin resistant cases were considered as MDR-TB. Results: Proportion of MDR-TB was 31.89% among retreatment TB cases. 56.75 % (n=21) of MDR-TB cases were between the age group of 21-40 years. Proportion of MDR-TB was higher among males (75.67%), married (59.45%) and rural dwellers 59.45%. Proportion was 61.76% among patients with BMI <16; 31.57% with BMI 16-16.99, 28.57% with BMI 17-18.49 and 5.7% with BMI 18.50-24.99. MDRTB proportion was less 29.72% (n=11) in patients with history of TB contact in family. Pulmonary TB including MDR-TB was more common among illiterates (37.83% among MDR). Conclusion: Proportion of MDR-TB was high among retreatment cases in north India. Among them low BMI and education status are modifiable factor and this study signifies that MDR-TB burden can be reduced by improving health and education status of patient.

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Prevalence and Factors Associated with Multidrug-Resistant Tuberculosis (MDR-TB) among Presumptive MDR-TB Patients in Tigray Region, Northern Ethiopia

Canadian Journal of Infectious Diseases and Medical Microbiology ◽

10.1155/2019/2923549 ◽

2019 ◽

Vol 2019 ◽

pp. 1-8 ◽

Cited By ~ 4

Author(s):

Kibriti Mehari ◽

Tsehaye Asmelash ◽

Haftamu Hailekiros ◽

Tewolde Wubayehu ◽

Hagos Godefay ◽

...

Keyword(s):

Odds Ratio ◽

Multidrug Resistant ◽

Sputum Smear ◽

Northern Ethiopia ◽

Crude Odds Ratio ◽

Factors Associated ◽

Resistant Tuberculosis ◽

Multidrug Resistant Tuberculosis ◽

Regional State ◽

Mdr Tb

Background. Tuberculosis (TB) is one of the major public health problems. There are alarming reports of increasing multidrug-resistant tuberculosis (MTR-TB) from various parts of the globe, including Ethiopia. This study was designed to determine the prevalence and factors associated with MDR-TB among presumptive MDR-TB cases in Tigray Regional State, Ethiopia. Methods. A cross-sectional study was conducted in Tigray Regional State from 2015 to 2016. Two hundred sputum samples were collected, transported, processed using 2% N-acetyl-L-cysteine-sodium hydroxide, and cultured in LJ medium. Besides, the microscopic examination was performed after ZN staining. Moreover, drug susceptibility test was done using molecular line probe assay. Descriptive statistics and binary and multivariable logistic regression were done. A statistical test was regarded as significant when the P value was <0.05. Results. The prevalence of MDR-TB was found to be 18.5%. About one-fourth (26.5%) of the study participants had sputum smear positive for acid-fast bacilli (AFB). TB culture was positive in 37% of the samples, and rifampicin mono-resistant cases accounted for 3.5% of the presumptive MDR-TB cases. Three (1.5%) were new MDR-TB cases, while the rest had been treated previously for TB. Most (63.5%) of the MDR-TB cases were from 15 to 44 years of age. Age was associated with MDR-TB with a crude odds ratio of 1.06 (CI: 1.02–1.10) and adjusted odds ratio of 1.06 (CI: 1.00–1.11). Conclusions. The prevalence of MDR-TB was found to be high. Preventive measures should be taken to prevent the transmission of MDR-TB in the community.

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Molecular Characterization of Multidrug-Resistant Mycobacterium tuberculosis Isolated in Nepal

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.06418-11 ◽

2012 ◽

Vol 56 (6) ◽

pp. 2831-2836 ◽

Cited By ~ 38

Author(s):

Ajay Poudel ◽

Chie Nakajima ◽

Yukari Fukushima ◽

Haruka Suzuki ◽

Basu Dev Pandey ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Promoter Region ◽

Multidrug Resistant ◽

Content Type ◽

Molecular Tests ◽

Resistant Tuberculosis ◽

Multidrug Resistant Tuberculosis ◽

Mdr Tb ◽

Substitution Mutations ◽

Drug Resistant Strains

ABSTRACTDespite the fact that Nepal is one of the first countries globally to introduce multidrug-resistant tuberculosis (MDR-TB) case management, the number of MDR-TB cases is continuing to rise in Nepal. Rapid molecular tests applicable in this setting to identify resistant organisms would be an effective tool in reversing this trend. To develop such tools, information about the frequency and distribution of mutations that are associated with phenotypic drug resistance inMycobacterium tuberculosisis required. In the present study, we investigated the prevalence of mutations inrpoBandkatGgenes and theinhApromoter region in 158M. tuberculosisisolates (109 phenotypically MDR and 49 non-MDR isolates collected in Nepal) by DNA sequencing. Mutations affecting the 81-bp rifampin (RIF) resistance-determining region (RRDR) ofrpoBwere identified in 106 of 109 (97.3%) RIF-resistant isolates. Codons 531, 526, and 516 were the most commonly affected, at percentages of 58.7, 15.6, and 15.6%, respectively. Of 113 isoniazid (INH)-resistant isolates, 99 (87.6%) had mutations in thekatGgene, with Ser315Thr being the most prevalent (81.4%) substitution. Mutations in theinhApromoter region were detected in 14 (12.4%) INH-resistant isolates. The results from this study provide an overview of the current situation of RIF and INH resistance inM. tuberculosisin Nepal and can serve as a basis for developing or improving rapid molecular tests to monitor drug-resistant strains in this country.

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Genotypic diversity of multi- and pre-extremely drug-resistant Mycobacterium tuberculosis isolates from Morocco

PLoS ONE ◽

10.1371/journal.pone.0253826 ◽

2021 ◽

Vol 16 (7) ◽

pp. e0253826

Author(s):

Amal Oudghiri ◽

Ghizlane Momen ◽

Achraf Aainouss ◽

Amin Laglaoui ◽

My Driss El Messaoudi ◽

...

Keyword(s):

Genetic Diversity ◽

Mycobacterium Tuberculosis ◽

Genotypic Diversity ◽

Multidrug Resistant ◽

Drug Resistant ◽

Control Programs ◽

Resistant Tuberculosis ◽

Mdr Tb ◽

Database Clustering ◽

High Level

In Morocco, the prevalence of multidrug resistant tuberculosis (MDR-TB) continues to increase especially within previously treated cases; these MDR cases may evolve to extensively drug resistant tuberculosis (XDR-TB) raising major concern to TB control programs. From an epidemiological window, scarce informations are available about the genetic diversity of Mycobacterium tuberculosis (MTB) strains fueling these forms of resistance. The aim of this study was to assess to genetic diversity of MDR-MTB strains. Hence, this prospective study was conducted on patients diagnosed with MDR-TB at Pasteur Institute of Casablanca from 2010 to 2013. A total of 70 MDR-MTB isolates were genotyped by spoligotyping and 15-loci MIRU-VNTR methods. Spoligotyping generated four orphan patterns, five unique profiles whereas 61 strains were grouped in nine clusters (2 to 25 strains per cluster), the clustering rates being 87.1%. Subtyping by 15 loci MIRU-VNTR splitted all clusters already established by spoligotyping and generated 70 unique profiles not recognized in SITVIT2 database; clustering rate was equal to zero. HGDI analysis of 15 loci MIRU demonstrated that eight out of 15 loci were highly discriminant. Of note, all pre-XDR strains belongs to many clades, meaning that there no association between gyrA mutants and particular clade. Overall, the data generated by this study (i) describe the population structure of MDR MTBC in Morocco which is highly homogenous, (ii) confirm that TB in Morocco is almost exclusively transmitted by modern and evolutionary lineages with high level of biodiversity seen by MIRU, and (iii) validate the use of optimized 15-loci MIRU-VNTR format for future investigations in Morocco.

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Epidemiology of Rifampicin Resistant Tuberculosis and Common Mutations in rpoB Gene of Mycobacterium tuberculosis: A Retrospective Study from Six Districts of Punjab (India) Using Xpert MTB/RIF Assay

Journal of Laboratory Physicians ◽

10.4103/0974-2727.180789 ◽

2016 ◽

Vol 8 (02) ◽

pp. 096-100 ◽

Cited By ~ 4

Author(s):

Ramandeep Kaur ◽

Neerja Jindal ◽

Shilpa Arora ◽

Shajla Kataria

Keyword(s):

Mycobacterium Tuberculosis ◽

Surrogate Marker ◽

Rpob Gene ◽

Multidrug Resistant ◽

Resistance Pattern ◽

Resistance Mutations ◽

Resistant Tuberculosis ◽

Mdr Tb ◽

Previously Treated Patients ◽

User Friendly

ABSTRACT Background: Xpert MTB/RIF assay has revolutionized the diagnosis of tuberculosis (TB) by simultaneously detecting the bacteria and resistance to rifampicin (RIF), a surrogate marker for multidrug-resistant TB (MDR-TB) in <2 h. The RIF resistance pattern in Malwa region of Punjab, India, is not documented. Here, we report the epidemiology of RIF-resistant TB and mutations in rpoB gene of Mycobacterium tuberculosis (MTB). Materials and Methods: A total of 1612 specimens received between October 2013 and February 2015 were tested by Xpert MTB/RIF assay following manufacturer’s instructions. The results thus obtained were analyzed using SPSS version 20.0.0 (SPSS Inc., Chicago, IL, USA) statistical software. Result: RIF resistance was statistically higher in previously treated patients in comparison to the new patients (P = 0.006) and in patients with acid fast-Bacilli (AFB) positive smears to AFB-negative smears (P = 0.048). RIF resistance mutations in 130 specimens revealed frequency of E 73/130 (56%), B 28/130 (21.5%), D 18/130 (13.8%), A 11/130 (8.4%), and C 1/130 (0.7%) while in one specimen, mutation combination, i.e., mutations associated with more than one probe (A and B both) was present. Conclusion: Xpert MTB/RIF assay is a user-friendly screening tool for detection of MTB and RIF resistance from suspected TB/MDR cases in a shorter period of time. It could also serve as a useful technique to have simultaneous preliminary information regarding the mutation pattern of RIF resistance in MTB isolates.

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RAPID DETECTION OF MULTI DRUG RESISTANCE AMONG MULTI DRUG RESISTANT TUBERCULOSIS SUSPECTS USING LINE PROBE ASSAY

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2017.v10i1.14341 ◽

2016 ◽

Vol 10 (1) ◽

pp. 131

Author(s):

Sanjeev Saini ◽

Manoj Kumar Dubey ◽

Uma Bhardwaj ◽

M Hanif ◽

Chopra Kk ◽

...

Keyword(s):

Fluorescent Microscopy ◽

Multi Drug Resistance ◽

Sputum Smear ◽

Drug Resistant ◽

Line Probe Assay ◽

Drug Resistant Tuberculosis ◽

Resistant Tuberculosis ◽

Multi Drug Resistant Tuberculosis ◽

Mdr Tb ◽

Probe Assay

ABSTRACTObjective: GenoType MTBDRplus line probe assay (LPA) is developed for performing drug susceptibility testing (DST) for Rifampicin (RIF) andisoniazid in sputum specimens from smear-positive pulmonary tuberculosis (TB) patients and revised national TB control Programme (RNTCP)has endorsed LPA for the diagnosis of multi drug resistant TB (MDR-TB). This study was conducted to assess the potential utility of LPA for MDR-TBpatient management.Methods: MDR-TB suspects under RNTCP PMDT criteria C referred from different districts in Delhi state were included in the study January 2013 toDecember 2014. Sputum specimens found acid-fast bacilli positive by fluorescent microscopy were processed for LPA.Results: Out of 3062 specimens, 2055 (67.1%) MDR-TB suspects were read as positive and specimens from 1007 (32.9%) suspects were read asnegative in sputum smear microscopy. Out of 2019 specimens valid LPA results, 1427 were found to be pan-sensitive, 280 were MDR-TB, 40 were RIFmonoresistant, 183 were Isoniazid (INH) monoresistant, and 89 specimens were found negative for Mycobacterium tuberculosis.Conclusion: Routine use of LPA can substantially reduce the time to diagnosis of RIF and/or INH-resistant TB and can hence potentially enable earliercommencement of appropriate drug therapy and thereby facilitate prevention of further transmission of drug resistant strains.Keywords: Multi drug resistant tuberculosis, Line probe assay, Rifampicin, Isoniazid.

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Draft Genome Sequences of Two Drug-Resistant Mycobacterium tuberculosis Isolates from Myanmar

Genome Announcements ◽

10.1128/genomea.00850-16 ◽

2016 ◽

Vol 4 (5) ◽

Cited By ~ 1

Author(s):

Htin Lin Aung ◽

Thanda Tun ◽

Elizabeth Permina ◽

Wint Wint Nyunt ◽

Si Thu Aung ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Draft Genome ◽

Multidrug Resistant ◽

Drug Resistant ◽

Genome Sequences ◽

Health Concerns ◽

Resistant Tuberculosis ◽

Multidrug Resistant Tuberculosis ◽

Extensively Drug Resistant ◽

Mdr Tb

Multidrug-resistant tuberculosis (MDR-TB) and lately, extensively drug-resistant TB (XDR-TB) are increasing global health concerns. Here, we present the genome sequences of two MDR-TB isolates from Myanmar, one of 27 countries with a high MDR-TB burden, and describe a number of mutations consistent with these being XDR-TB isolates.

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