Line Probe Assay for Improved Tuberculosis Diagnosis and Detection of Isoniazid and Rifamicin Resistance

Chaburte, Chala (2015) Line Probe Assay for Improved Tuberculosis Diagnosis and Detection of Isoniazid and Rifamicin Resistance. Masters thesis, Addis Ababa University.

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Abstract

Introduction: A sensitive, rapid and accurate laboratory diagnostic method is essential for detection of tuberculosis and its drug resistance. However, using conventional techniques for detection of acidfast bacilli and mutations conferring resistance to anti-TB drugs have a number of draw backs. Objective: To evaluate the diagnostic ability of Genotype MTBDRplus version 2.0 line probe assay for diagnosis of Mycobacterium tuberculosis complex (MTBC) and detection of drug resistant tuberculosis from direct sputum specimens. Methods: A cross-sectional study was conducted in Addis Ababa regional laboratory from December 2013 to September 2014. A total of 96 drug resistant TB suspected patients were recruited from four hospitals and twenty eight health centers found in Addis Ababa. The specimens were analyzed in Addis Ababa regional laboratory and Armauer Hansen Research Institute. Culture by Lowenstein–Jensen medium, Genotype MTBDRplus version 2.0 line probe assay, pyrosequencing, spoligo-rifampicinisoniazid-typing and Gene Xpert MTB/RIF assay were performed. Results: The sensitivity and specificity of Genotype MTBDRplus version 2.0 line probe assay for detection of MTBC were 89.3% and 90.5%, respectively using culture method as a gold standard. The sensitivity and specificity of MTBDRplus were equivalent to that of Gene Xpert MTB/RIF assay in detecting MTBC in sputum samples. However, the performance of MTBDRplus in detecting resistance to rifampicin (47/66) was much higher than that of Gene Xpert MTB/RIF assay (30/66). The majority of mutations conferring resistance to rifampicin and isoniazid were found in codon S531L and S315T, respectively. The performances of MTBDRplus and spoligo-RIF-INH-typing were 96.2% concordant in detecting resistance to RIF, INH and multi-drug resistant tuberculosis. Conclusion and Recommendation: Genotype MTBDRplus version 2.0 line probe assay improves diagnosis of tuberculosis and detection of drug resistance TB in sputum specimen regardless of its smear status. The method is effective, time saving and relatively simple method for detection of drug resistance to INH and RIF provided that the weak or faint appearance of wild type 8 band without the presence of related mutation band is considered as normal during interpretation of the results for rifampicin resistance. Further studies should be conducted to assess factors affecting wild type 8. The manufacturer should evaluate and review the manual for interpretation of Genotype MTBDRplus version 2.0 line probe assay results.

Item Type: Thesis (Masters)
Uncontrolled Keywords: Drug resistance, Genotype MTBDRplus version 2.0 line probe assay, M. tuberculosis complex, Tuberculosis
Subjects: Q Science > Q Science (General)
Q Science > QH Natural history > QH301 Biology
Q Science > QR Microbiology
R Medicine > RA Public aspects of medicine > RA0421 Public health. Hygiene. Preventive Medicine
Divisions: Africana
Depositing User: Selom Ghislain
Date Deposited: 26 Sep 2018 10:45
Last Modified: 26 Sep 2018 10:45
URI: http://thesisbank.jhia.ac.ke/id/eprint/5593

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