scholarly journals Genetic mutations at rifampicin resistance-determining region of RpoB gene in conferring resistance to isolated multidrug-resistant tuberculosis strains

2018 ◽  
Vol 2 (4) ◽  
pp. 295
Author(s):  
PrasantaKumar Das ◽  
SomtirthaB Ganguly ◽  
Bodhisatya Mandal
Keyword(s):  
Rpob Gene ◽  
Multidrug Resistant ◽  
Rifampicin Resistance ◽  
Genetic Mutations ◽  
Resistant Tuberculosis ◽  
Multidrug Resistant Tuberculosis

HYBRIDIZATION-BASED NUCLEIC ACID AMPLIFICATION TEST TERHADAP CARTRIDGE-BASED NUCLEIC ACID AMPLIFICATION TEST TERKAIT MULTIDRUG-RESISTANT TUBERCULOSIS

2016 ◽  
Vol 21 (3) ◽  
pp. 237
Author(s):  
Ivana Agnes Sulianto ◽  
Ida Parwati ◽  
Nina Tristina ◽  
Agnes Rengga I
Keyword(s):  
Nucleic Acid ◽  
Rpob Gene ◽  
Multidrug Resistant ◽  
Nucleic Acid Amplification Test ◽  
Nucleic Acid Amplification ◽  
Isoniazid Resistance ◽  
Resistant Tuberculosis ◽  
Multidrug Resistant Tuberculosis ◽  
Mdr Tb ◽  
Low Sensitivity

Indonesia has high burden of multidrug-resistant tuberculosis (MDR-TB). Cartridge-based nucleic acid amplification test (CB-NAAT), which is recommended as a diagnostic method of MDR-TB by World Health Organization, is faster in achieving the result. This method determines MDR-TB only from the rifampisin resistance, by detecting mutations that occur on the 81 bp hot-spot region of the rpoB gene. The isoniazid resistance is not included in the determination of MDR-TB by this method. Hybridization-based NAAT (HB-NAAT) detects MDR-TB not only from the rifampisin resistance (codon 526 and 531 rpoB gene), but also from the isoniazid resistance (codon 315 katG gene). The aim of this study was to know the validity of the HB-NAAT in detecting MDR-TB using sputum with CB-NAAT as the gold standard in a diagnostic study. All of 51 sputums were collected during June 2013 from patients suspected pulmonary MDR-TB at Dr. Hasan Sadikin General Hospital. The result of CB-NAAT were 16 MDR-TB, 12 TB non MDR, and 23 non TB. HB-NAAT examination results were 3 MDR-TB, 25 TB non MDR (3 RMR, 6 IMR, 16 susceptible) and 23 non TB. The sensitivity of HB-NAAT was 18.75% and specificity 100%. Low sensitivity values may due to the high mutation variations in the samples. So it could not be detected only by codons 526 and 531 for rifampisin resistance. For the detection of isoniazid resistance, HB-NAAT have optimal primer at low concentrations and it also need more than katG genes to detect isoniazid resistance. Based on this study, it can be conclued, that HBNAAT has low sensitivity but high specificity in the detecting MDR-TB.

scholarly journals Comparison of Interpretation between Pyrosequencing and Xpert MTB/RIF Assay in Multidrug-Resistant Tuberculosis

2020 ◽  
Vol 52 (4) ◽  
Author(s):  
Linda Choerunnisa ◽  
Ida Parwati ◽  
Coriejati Rita ◽  
Anna Tjandrawati ◽  
Lidya Chaidir
Keyword(s):  
Gene Mutations ◽  
Rpob Gene ◽  
Multidrug Resistant ◽  
Detection Methods ◽  
Isoniazid Resistance ◽  
Resistant Tuberculosis ◽  
Position Detection ◽  
Multidrug Resistant Tuberculosis ◽  
Mdr Tb ◽  
Codon Mutation

Indonesia is one of the countries with the highest multidrug-resistant tuberculosis cases in the world. Rapid molecular test using the Xpert MTB/RIF assay is one of the detection methods for MDR-TB. Early detection of MDR-TB is crucial for early initiation of treatment. However, Xpert MTB/RIF assay only detects the rpoB gene mutations associated with Rifampicin resistance. Recently, WHO recommends the use of Pyrosequencing, a DNA sequencing method that can detect not only the rpoB gene but also katG and/or inhA gene mutations associated with Isoniazid resistance. The aims of this study were to compare the interpretation between the two methods and to determine the differences in codon mutation position detection of the rpoB gene and mutation detection of the katG and/or inhA gene. This was a cross-sectional comparative observational study on patients ≥18 years old interpreted as RR-TB patients based on Xpert MTB/RIF assay results who had not received MDR-TB drugs at Dr. Hasan Sadikin General Hospital, Bandung, Indonesia. Results showed there were 40 Rifampicin-resistant TB subjects interpreted by Xpert MTB/RIF assay while Pyrosequencing interpreted 30 MDR-TB, 9 RR-TB and one Isoniazid-resistant TB subjects in January - February 2020. The detection of rpoB gene codon mutation position between Xpert MTB/RIF assay and Pyrosequencing methods was not significantly different (p=0.389). Pyrosequencing had detected 27 katG gene mutations, 3 inhA gene mutations, one katG and inhA gene mutation. To conclude, Pyrosequencing can be used for accurate detection of Rifampicin and Isoniazid resistance in MDR-TB.

APPLICATION OF REALTIME PCR FOR DETECTING M.TUBERCULOSIS RESISTANT TO RIFAMPICIN AND ISONIAZID

2015 ◽  
pp. 110-116
Author(s):  
Viet Quynh Tram Ngo ◽  
Thanh Quang Nguyen ◽  
Thi Hai Duong Huynh ◽  
Nu Xuan Thanh Le
Keyword(s):  
Rpob Gene ◽  
Multidrug Resistant ◽  
Katg Gene ◽  
Culture Methods ◽  
Realtime Pcr ◽  
Conventional Culture ◽  
Resistant Tuberculosis ◽  
Multidrug Resistant Tuberculosis ◽  
Resistant Strains ◽  
Recurrent Tuberculosis

Introduction: Conventional culture methods for antibiotic susceptibility testing for M. tuberculosis may take days to several weeks. Use of realtime PCR for rapid detection of drug and multidrug resistant tuberculosis could facilitate early initiation of appropriate anti-tubercular treatment regimen thereby interrupting transmission. Objectives: 1. Determine prevalence of rifampicin-, isoniazid- and multidrug-resistant tuberculosis strain among the patients with recurrent tuberculosis. Xác định tỷ lệ các đột biến kháng rifampicin và isoniazid trong các chủng đề kháng phenotype 2. Determine proportion of the mutations concern to tuberculosis strains resistant to rifampicin và isoniazid. Methods: Doing susceptibility test by MODS assay for 50 tuberculosis strains isolated from patients with recurrent tuberculosis, realtime PCR was performed by using MTB Real-TM Resistance 4 kit to detect mutations in codon 531 of the rpoB gene related to rifampicine and mutations in codon 315 of the katG gene or in codon 209 of the inhA gene related to isoniazid. Results: Among tuberculosis strains isolated from patients with recurrent tuberculosis, there were 8% strains monoresistant to rifampicin and 60% multidrug-resistant strains. The Sacace MTB Real-TM resistance 4 kit detected 80% tuberculosis strains resistant to rifampicine with the mutation in rpoB gene codon 531 and 86,1% tuberculosis strains resistant to isoniazid with the mutation in katG gene codon 315 among the strains determined by MODS assay. Conclusion: The Sacace MTB Real-TM resistance 4 kit can’t detect rifampicine-/ isoniazid - resistant tuberculosis strains which have mutations in other codons of the above gene. Key words: M. tuberculosis, rifampicine-resistance, isoniazid-resistance, MODS, realtime PCR

scholarly journals Proportion of Rifampicin Resistance in Sputum Smear Positive Pulmonary Tuberculosis with Diabetes Mellitus in Urban Bengaluru - A Cross-Sectional Study

2021 ◽  
Vol 8 (23) ◽  
pp. 1888-1893
Author(s):  
Srinivasa Kaligonahalli Venkataramanappa ◽  
Jeevan Basavaraj
Keyword(s):  
Diabetes Mellitus ◽  
Type 2 Diabetes ◽  
Pulmonary Tuberculosis ◽  
Multidrug Resistant ◽  
Rifampicin Resistance ◽  
Sputum Smear ◽  
Diabetic Patients ◽  
Resistant Tuberculosis ◽  
Multidrug Resistant Tuberculosis

BACKGROUND Pulmonary tuberculosis (PTB) still remains a global public health problem. Diabetes along with poor glycaemic control leads to an immune compromised state. Type 2 diabetes (DM) is a strong risk factor for tuberculosis (TB) and is associated with a slower response to TB treatment and a higher mortality rate. Objective of the study was to determine the prevalence of rifampicin resistance in pulmonary tuberculosis in patients with DM. METHODS The study was conducted in General Medicine Department, Dr. B.R. Ambedkar Medical College, Bengaluru, Karnataka, from July 2018 to December 2019. Sputum was collected from a total of 100 patients who were sputum smear positive for acid fast bacilli (AFB). Sample was collected and was subjected to GeneXpert testing for the evaluation of resistance against rifampicin. RESULTS In our study the incidence of rifampicin resistance was 45.16 % in diabetes and 5.79 % in non-diabetes which was statistically significant ( 2 -1.1; p-0.01) between rifampicin resistance and diabetic patients’ status with active TB in South Karnataka, India. This result supports previous research showing a higher multidrug-resistant tuberculosis risk in tuberculosis & diabetes patients relative to those with no diabetes. Additionally, correlation of other findings between type 2 diabetes & multidrug-resistant tuberculosis as well as delayed time to sputum smear conversion were confirmed. CONCLUSIONS The results showed correlation between diabetes & rifampicin resistance and probably suggest the necessity of integrated diabetes and tuberculosis surveillance programs in South Karnataka, India. KEYWORDS Rifampicin, Tuberculosis, Diabetes Mellitus

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