PREVALANCE OF GENETIC MUTATION ASSOSIATED WITH (RPOB) RIFAMPICIN RESISTANT AMONG THE TREATED TUBERCULOSIS PATIENTS

2020 ◽  
Vol 11 (12) ◽  
pp. 235-268
Author(s):  
Girija Vasudevan ◽  
Muthuraj M ◽  
Antony V samrot
Keyword(s):  
Drug Susceptibility ◽  
Genetic Mutation ◽  
Drug Susceptibility Testing ◽  
Nucleic Acid Amplification ◽  
Drug Resistant ◽  
Tuberculosis Patients ◽  
Resistant Tuberculosis ◽  
Resource Limited ◽  
Probe Assay ◽  
Solid Liquid

Tuberculosis (TB) is an infectious disease caused predominantly by Mycobacterium tuberculosis. India accounts for one fourth of the global TB burden. Drug-resistant tuberculosis (TB) undermines control efforts and its burden is poorly understood in resource-limited settings. Drug resistant TB is a laboratory based diagnosis and is performed either by phenotypic drug susceptibility testing using solid/liquid culture or genotypic testing for detection of resistance by Line Probe Assay/Cartridge based Nucleic Acid Amplification tests.

scholarly journals Compatibility of a novel filter paper-based bio-safe sputum transport kit with Line Probe Assay for diagnosing drug-resistant tuberculosis: a single-site evaluation study

2021 ◽  
pp. 00137-2021
Author(s):  
Divya Anthwal ◽  
Rakesh Kumar Gupta ◽  
Ritu Singhal ◽  
Manpreet Bhalla ◽  
Ajoy Kumar Verma ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Dna Extraction ◽  
Cost Effective ◽  
Evaluation Study ◽  
Drug Susceptibility ◽  
Drug Susceptibility Testing ◽  
Drug Resistant ◽  
Resistant Tuberculosis ◽  
Site Evaluation ◽  
Probe Assay

BackgroundNear-patient access to appropriate tests is a major obstacle for the efficient diagnosis of Tuberculosis (TB) and associated drug resistance.MethodsWe recently developed the “TB Concentration & Transport” kit for bio-safe, ambient-temperature transportation of dried sputum on Trans-Filter, and the “TB DNA Extraction” kit for DNA extraction from Trans-Filter for determining drug resistance by DNA sequencing. In the present study, we evaluated the compatibility of Kit-extracted DNA with Hain's Line Probe Assays (LPAs), which are endorsed by National TB programmes for the detection of drug resistance in sputum collected from presumptive Multi-drug resistant TB patients (n=207).ResultsTrans-Filter-extracted DNA was seamlessly integrated with the LPA protocol (Kit-LPA). The sensitivity of Kit-LPA for determining drug resistance was 83.3% for rifampicin (95% Confidence Interval [CI]: 52, 98%), 77.7% for isoniazid (95% CI: 52, 94%), 85.7% for fluoroquinolones (95% CI: 42, 100%) and 66.6% for aminoglycosides (95% CI: 9, 99%), with a specificity range of 93.7% (95% CI: 87, 97) to 99.1% (95% CI: 95, 100) using phenotypic drug susceptibility testing (DST) as a reference standard. A high degree of concordance was noted between results obtained from Kit-LPA and LPA [99% to 100% (κ value: 0.83–1.0)].ConclusionsThis study demonstrates successful integration of our developed kits with LPA. The adoption of these kits across Designated Microscopy Centres in India can potentially overcome the existing challenge of transporting infectious sputum at controlled temperature to centralised testing laboratories and can provide rapid near-patient cost-effective “Universal DST” services to TB subjects residing in remote areas.

scholarly journals Drug Resistant Tuberculosis in the Northern Region of Zambia: A Retrospective Study

2021 ◽  
Vol 2 ◽  
Author(s):  
Ngula Monde ◽  
Mildred Zulu ◽  
Mathias Tembo ◽  
Ray Handema ◽  
Musso Munyeme ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Susceptibility Testing ◽  
Drug Susceptibility ◽  
Drug Susceptibility Testing ◽  
Multi Drug Resistance ◽  
Drug Resistant ◽  
Drug Resistant Tuberculosis ◽  
Tuberculosis Patients ◽  
Resistant Tuberculosis ◽  
Multi Drug Resistant Tuberculosis

BackgroundZambia like many countries in sub-Saharan Africa is affected with drug resistant tuberculosis. However, the drug resistant tuberculosis situation over the years has not been described in various regions of the country. Consequently, this study aims to determine the drug resistant tuberculosis burden in northern regions of Zambia over a four-year period based on data generated from a Regional Tuberculosis Reference Laboratory.MethodTwo hundred and thirty two (232) Tuberculosis Drug Susceptibility Testing results over a four-year period (2016-2019) were reviewed. Data was collected from tuberculosis registers and patient request forms and entered into a pre-tested standardized checklist and later entered in Excel Computer software. Double blinded checking was done by two independent data clerks to minimize duplication of cases. Cleaned data was then imported in R programme for analysis. Bivariant and descriptive statistics were performed and reported.ResultsOf 232 Drug Susceptibility Testing results, 90.9% were drug resistant TB while 9% were drug susceptible. Fifty three percent (53%) of these were multi-drug resistant Tuberculosis and 32% were confirmed as Rifampicin Mono-resistance. Only 1.7% of the Multi-drug resistant Tuberculosis patients were Pre-extensively drug-resistant Tuberculosis. Copperbelt province had the largest proportion (46.0%) of multi-drug resistant tuberculosis patients followed by Luapula (8.1%) and North-Western (4.7%) provinces. In new and previously treated patients, the proportion of Multi-drug resistant tuberculosis was 71.8% and 28.7% respectively. History of previous anti-tuberculosis treatment and treatment failure were associated with multi-drug resistance TB.Conclusion and RecommendationThis study has shown a small increase in the proportions of drug resistant tuberculosis cases over the four years under review with high rates being recorded on the Copperbelt Province. Previous treatment to first line TB treatment and treatment failure were associated with development of Multi-drug resistance. We therefore recommend strengthened routine laboratory surveillance and improved case management of multi-drug resistant tuberculosis patients in the region.

scholarly journals Rapid Detection of Rifampicin and Isoniazid Resistant Mycobacterium tuberculosis Using Genotype MTBDRplus Assay in Nepal

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Bijay Kumar Sharma ◽  
Shiva Bhandari ◽  
Bhagwan Maharjan ◽  
Bhawana Shrestha ◽  
Megha Raj Banjara
Keyword(s):  
Drug Susceptibility ◽  
Multidrug Resistant ◽  
Drug Susceptibility Testing ◽  
Drug Resistant ◽  
Mutational Frequency ◽  
Resistant Tuberculosis ◽  
Mdr Tb ◽  
Proportion Method ◽  
Genotype Mtbdrplus ◽  
Probe Assay

Rapid line probe assay (LPA) can be a practical and rapid alternative to the slow conventional phenotypic drug susceptibility testing (DST) for detection of drug resistant tuberculosis (TB). The purpose of this study is to determine the diagnostic accuracy of Genotype MTBDRplus, LPA for TB, and compare its performance with conventional DST. A total of 54 culture samples were analyzed for DST using both conventional proportion method and MTBDRplus, where conventional DST identified 43 isolates (79.6%) as drug resistant. Among these 43 drug resistant isolates, 30 isolates (69.7%) were found to be multidrug resistant (MDR). Of all observed mutations using MTBDRplus, codon 531 of rpoB gene and codon 315 of katG gene were found to have highest mutational frequency for RIF resistance (64.7%) and INH resistance (96.8%), respectively. In the present study, MTBDRplus assay was shown to have excellent specificity (100%) for both RIF and INH resistance while sensitivity of the assay was little lower with value of 89.4% for RIF resistance and 91.4% for INH resistance. Therefore, the assay can be a rapid, reliable, and promising molecular test for early detection of MDR-TB in Nepal.

Uptake of universal drug susceptibility testing among people with TB in a south Indian district: How are we faring?

2021 ◽  
Author(s):  
Rajani Ranganath ◽  
Hemant Deepak Shewade ◽  
Abdul K Bahadur ◽  
Venkatesh Naik ◽  
Sharath Burugina Nagaraja ◽  
...  
Keyword(s):  
Susceptibility Testing ◽  
Secondary Data ◽  
Drug Susceptibility ◽  
Drug Susceptibility Testing ◽  
Nucleic Acid Amplification ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Public Facilities ◽  
South Indian ◽  
Probe Assay

Abstract Background India implements universal drug susceptibility testing (UDST) using rapid genotypic tests (cartridge-based nucleic acid amplification test CBNAAT - and line probe assay - LPA). to bridge the gap of diagnosis of multidrug/rifampicin-resistant TB. There is limited evidence assessing the implementation of UDST in India. We assessed the implementation among people with pulmonary TB notified from public facilities in October 2019 from Raichur (Karnataka), India. Methods A cohort study involving secondary data in routine programme settings was conducted. All people with TB underwent a rapid genotypic DST for rifampicin resistance followed by first line-LPA (FL-LPA) if sensitive and second line-LPA (SL-LPA) if resistant. Results Of 217 people, 15.7% (n=34) did not undergo rapid genotypic DST. Of 135 who were rifampicin-sensitive detected on CBNAAT, 68.1% (n=92) underwent FL-LPA, and out of the six rifampicin-resistant cases, 66.7% (n=4) underwent SL-LPA. Overall, 65.4% (142/217) completed the UDST algorithm. Children (aged <15 y) and people with bacteriological non-confirmation on microscopy were less likely to undergo rapid genotypic DST. Of 183 patients who underwent both rapid genotypic DST and sputum smear microscopy, 150 were bacteriologically confirmed and, of them, 9 (6%) were ‘rapid DST-negative’. Conclusion We found gaps at various steps. There were a significant number of ‘rapid DST-negative, smear-positive’ patients.

scholarly journals Speeding up the diagnosis of multidrug-resistant tuberculosis in a high-burden region with the use of a commercial line probe assay

2019 ◽  
Vol 45 (2) ◽  
Author(s):  
Angela Pires Brandao ◽  
Juliana Maira Watanabe Pinhata ◽  
Rosangela Siqueira Oliveira ◽  
Vera Maria Neder Galesi ◽  
Helio Hehl Caiaffa-Filho ◽  
...  
Keyword(s):  
Susceptibility Testing ◽  
Drug Susceptibility ◽  
Multidrug Resistant ◽  
Drug Susceptibility Testing ◽  
Reference Laboratory ◽  
Line Probe Assay ◽  
Resistant Tuberculosis ◽  
Multidrug Resistant Tuberculosis ◽  
Commercial Line ◽  
Probe Assay

ABSTRACT Objective: To evaluate the rapid diagnosis of multidrug-resistant tuberculosis, by using a commercial line probe assay for rifampicin and isoniazid detection (LPA-plus), in the routine workflow of a tuberculosis reference laboratory. Methods: The LPA-plus was prospectively evaluated on 341 isolates concurrently submitted to the automated liquid drug susceptibility testing system. Results: Among 303 phenotypically valid results, none was genotypically rifampicin false-susceptible (13/13; 100% sensitivity). Two rifampicin-susceptible isolates harboured rpoB mutations (288/290; 99.3% specificity) which, however, were non-resistance-conferring mutations. LPA-plus missed three isoniazid-resistant isolates (23/26; 88.5% sensitivity) and detected all isoniazid-susceptible isolates (277/277; 100% specificity). Among the 38 (11%) invalid phenotypic results, LPA-plus identified 31 rifampicin- and isoniazid-susceptible isolates, one isoniazid-resistant and six as non-Mycobacterium tuberculosis complex. Conclusions: LPA-plus showed excellent agreement (≥91%) and accuracy (≥99%). Implementing LPA-plus in our setting can speed up the diagnosis of multidrug-resistant tuberculosis, yield a significantly higher number of valid results than phenotypic drug susceptibility testing and provide further information on the drug-resistance level.

scholarly journals Sensititre MycoTB Plate Compared to Bactec MGIT 960 for First- and Second-Line Antituberculosis Drug Susceptibility Testing in Tanzania: a Call To Operationalize MICs

2015 ◽  
Vol 59 (11) ◽  
pp. 7104-7108 ◽  
Author(s):  
Scott K. Heysell ◽  
Suporn Pholwat ◽  
Stellah G. Mpagama ◽  
Saumu J. Pazia ◽  
Happy Kumburu ◽  
...  
Keyword(s):  
Susceptibility Testing ◽  
Acquired Resistance ◽  
Drug Susceptibility ◽  
Drug Susceptibility Testing ◽  
Drug Resistant ◽  
Second Line ◽  
Drug Resistant Tuberculosis ◽  
Content Type ◽  
Resistant Tuberculosis ◽  
Extensively Drug Resistant

ABSTRACTMIC testing forMycobacterium tuberculosisis now commercially available. Drug susceptibility testing by the MycoTB MIC plate has not been directly compared to that by the Bactec MGIT 960. We describe a case of extensively drug-resistant tuberculosis (XDR-TB) in Tanzania where initial MIC testing may have prevented acquired resistance. From testing on archived isolates, the accuracy with the MycoTB plate was >90% for important first- and second-line drugs compared to that with the MGIT 960, and clinically useful quantitative interpretation was also provided.

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