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.

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 Prevalence and Molecular Characteristics of Drug-Resistant Mycobacterium tuberculosis in Hunan, China

2014 ◽  
Vol 58 (6) ◽  
pp. 3475-3480 ◽  
Author(s):  
Li-li Zhao ◽  
Yan Chen ◽  
Zhong-nan Chen ◽  
Hai-can Liu ◽  
Pei-lei Hu ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Drug Susceptibility ◽  
Drug Susceptibility Testing ◽  
Hunan Province ◽  
Molecular Characteristics ◽  
Drug Resistant ◽  
Content Type ◽  
Resistant Tuberculosis ◽  
Frequent Mutations ◽  
Almost All

ABSTRACTTo determine the prevalence and molecular characteristics of drug-resistant tuberculosis in Hunan province, drug susceptibility testing and spoligotyping methods were performed among 171M. tuberculosisisolates. In addition, the mutated characteristics of 12 loci, includingkatG,inhA,rpoB,rpsL, nucleotides 388 to 1084 of therrsgene [rrs(388–1084)],embB,pncA,tlyA,eis, nucleotides 1158 to 1674 of therrsgene [rrs(1158–1674)],gyrA, andgyrB, among drug-resistant isolates were also analyzed by DNA sequencing. Our results indicated that the prevalences of isoniazid (INH), rifampin (RIF), streptomycin (SM), ethambutol (EMB), pyrazinamide (PZA), capreomycin (CAP), kanamycin (KAN), amikacin (AKM), and ofloxacin (OFX) resistance in Hunan province were 35.7%, 26.9%, 20.5%, 9.9% 15.2%, 2.3%, 1.8%, 1.2%, and 10.5%, respectively. The previously treated patients presented significantly increased risks for developing drug resistance. The majority ofM. tuberculosisisolates belonged to the Beijing family. Almost all the drug resistance results demonstrated no association with genotype. The most frequent mutations of drug-resistant isolates werekatGcodon 315 (katG315),inhA15,rpoB531,rpoB526,rpoB516,rpsL43,rrs514,embB306,pncA96,rrs1401,gyrA94, andgyrA90. These results contribute to the knowledge of the prevalence of drug resistance in Hunan province and also expand the molecular characteristics of drug resistance in China.

scholarly journals Drug susceptibility testing and mortality in patients treated for tuberculosis in high-burden countries

2018 ◽  
Author(s):  
Kathrin Zürcher ◽  
Marie Ballif ◽  
Lukas Fenner ◽  
Sonia Borrell ◽  
Peter M. Keller ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Susceptibility Testing ◽  
Drug Susceptibility ◽  
Drug Susceptibility Testing ◽  
Reference Laboratory ◽  
Drug Resistant ◽  
Hiv Status ◽  
Resistant Tuberculosis ◽  
Resistant Strains ◽  
Drug Resistant Strains

ABSTRACTBackgroundDrug resistance and HIV co-infection are challenges for the global control of tuberculosis.MethodsWe collected Mycobacterium tuberculosis isolates from adult patients in Côte d’Ivoire, Democratic Republic of the Congo, Kenya, Nigeria, South Africa, Peru, and Thailand, stratified by HIV status and tuberculosis drug resistance. Molecular or phenotypic drug susceptibility testing (DST) was done locally and at the Swiss tuberculosis reference laboratory. We examined mortality during treatment according to DST results and treatment adequacy in logistic regression models adjusting for sex, age, sputum microscopy and HIV status.Findings634 tuberculosis patients were included; median age was 33.2 years, 239 (37.7%) were female, 272 (42.9%) HIV-positive and 69 (10.9%) patients died. Based on the reference laboratory DST, 394 (62.2%) strains were pan-susceptible, 45 (7.1%) mono-resistant, 163 (25.7%) multidrug-resistant (MDR-TB), and 30 (4.7%) had pre-extensive or extensive drug resistance (pre-XDR/XDR-TB). Results of reference and local laboratories were discordant in 121 (19.1%) cases, corresponding to a sensitivity of 84.3% and a specificity of 90.8%. In patients with drug-resistant tuberculosis, discordant results were associated with increased mortality (risk ratio 1.81; 95% CI 1.07-3.07). In logistic regression, compared to adequately treated patients with pan-susceptible strains, the adjusted odds ratio for death was 4.23 (95% CI 2.16-8.29) for adequately treated patients with drug-resistant strains and 21.54 (95% CI 3.36-138.1) for inadequately treated patients with drug-resistant strains. HIV status was not associated with mortality.InterpretationUsing a reference laboratory standard, inaccurate DST leading to inappropriate treatment of drug-resistant tuberculosis, but not HIV infection, contributed to mortality.

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.

scholarly journals Evaluation of Nitrate Reductase Assay (NRA) for rapid detection of druc resistant tuberculosis at National Tuberculosis Centre, Nepal

1970 ◽  
Vol 7 (1) ◽  
pp. 26-30
Author(s):  
PK Mandal ◽  
S Basnyat ◽  
DK Khadka ◽  
DR Bhatta
Keyword(s):  
Drug Resistance ◽  
Nitrate Reductase ◽  
Sensitivity And Specificity ◽  
Drug Susceptibility ◽  
Drug Resistant ◽  
Drug Resistant Tuberculosis ◽  
Resistant Tuberculosis ◽  
National Tuberculosis ◽  
Nitrate Reductase Assay ◽  
Hiv Aids

Background: Treatment of drug-resistant tuberculosis is often based on drug susceptibility testing results. Thus simple, rapid and economic test is very important for diagnosis of drug resistant tuberculosis and such method aids in TB control effectively. One such method is a Nitrate Reductase Assay (NRA). Objective: To evaluate feasibility and performance of Nitrate Reductase Assay in the screening of drug-resistant tuberculosis. Setting: National Tuberculosis Centre and SAARC TB and HIV/AIDS Centre, Thimi, Bhaktapur, Nepal from April 2008 to March 2009. Methods: A prospective study comparing the sensitivity and specificity of the Nitrate Reductase Assay with the gold standard Lowenstein Jensen proportion method in determining drug susceptibility pattern to four primary anti-tubercular drugs i.e. isoniazid, rifampicin, streptomycin and ethambutal among clinical isolates.Results: Among 121 specimens , the sensitivity and specificity of the Nitrate Reductase Assay for detection of Isoniazid resistance was 100% and 91%, for rifampicin was 100% and 98.95%, for streptomycin was 96% and 91.66% and for ethambutal was 100% and 98% respectively.Conclusions: The Nitrate Reductase Assay is sensitive and specific enough for the detection of drug resistant tuberculosis. It is rapid, easy to use and inexpensive, making it suitable for developing countries. Its usefulness for national drug resistance surveys should be assessed. Keywords: diagnosis; drug resistance; sensitivity; specificity; NRADOI: 10.3126/saarctb.v7i1.3960SAARC J. TUBER. LUNG DIS. HIV/AIDS 2010 VII(1) 26-30

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 Evaluation of MTBDRplusand MTBDRslin Detecting Drug-Resistant Tuberculosis in a Chinese Population

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Wei Lu ◽  
Yan Feng ◽  
Jianming Wang ◽  
Limei Zhu
Keyword(s):  
Drug Resistance ◽  
Chinese Population ◽  
Sequencing Analysis ◽  
Drug Resistant ◽  
Line Probe Assay ◽  
Drug Resistant Tuberculosis ◽  
Additional Tool ◽  
Resistant Tuberculosis ◽  
Probe Assay ◽  
Mycobacteria Tuberculosis

Background. This study aims to evaluate GenoType MTBDRplusand GenoType MTBDRslfor their ability to detect drug-resistant tuberculosis in a Chinese population.Methods. We collected 112Mycobacteria tuberculosisstrains from Jiangsu province, China. The conventional DST and line probe assay were used to detect drug resistance to rifampicin (RFP), isoniazid (INH), ofloxacin (OFX), kanamycin (Km), and ethambutol (EMB).Results. The sensitivity and specificity were 100% and 50% for RFP and 86.11% and 47.06% for INH, respectively. The most common mutations observed in MTBDRpluswererpoBWT8 omission + MUT3 presence,katGWT omission + MUT1 presence, andinhAWT1 omission + MUT1 presence. For drug resistance to OFX, Km, and EMB, the sensitivity of MTBDRslwas 94.74%, 62.50%, and 58.82%, respectively, while the specificity was 92.59%, 98.81%, and 91.67%, respectively. The most common mutations weregyrAWT3 omission + MUT3C presence,rrsMUT1 presence,embBWT omission + MUT1B presence, andembBWT omission + MUT1A presence. Sequencing analysis found several uncommon mutations.Conclusion. In combination with DST, application of the GenoType MTBDRplusand GenoType MTBDRslassays might be a useful additional tool to allow for the rapid and safe diagnosis of drug resistance to RFP and OFX.

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