Multi-Fluorescence Real-Time PCR Assay for Detection of RIF and INH Resistance of M. tuberculosis

Failure to early detect multidrug-resistant tuberculosis (MDR-TB) results in treatment failure and poor clinical outcomes, and highlights the need to rapidly detect resistance to rifampicin (RIF) and isoniazid (INH). In Multi-Fluorescence quantitative Real-Time PCR (MF-qRT-PCR) assay, 10 probes labe...

Full description

Saved in:
Bibliographic Details
Published inFrontiers in microbiology Vol. 7; p. 618
Main Authors Peng, Jingfu, Yu, Xiaoli, Cui, Zhenling, Xue, Wenfei, Luo, Ziyi, Wen, Zilu, Liu, Minghua, Jiang, Danqing, Zheng, Heping, Wu, Hai, Zhang, Shulin, Li, Yao
Format Journal Article
LanguageEnglish
Published Switzerland Frontiers Media S.A 29.04.2016
Subjects
Drug resistanc
Isoniazid
Mycobacterium tuberculosis
Public Health
quantitative real-time PCR
rifampicin
rifampicin
isoniazid
drug resistance
Mycobacterium tuberculosis
quantitative real-time PCR
Online AccessGet full text

Cover

More Information
Summary:Failure to early detect multidrug-resistant tuberculosis (MDR-TB) results in treatment failure and poor clinical outcomes, and highlights the need to rapidly detect resistance to rifampicin (RIF) and isoniazid (INH). In Multi-Fluorescence quantitative Real-Time PCR (MF-qRT-PCR) assay, 10 probes labeled with four kinds of fluorophores were designed to detect the mutations in regions of rpoB, katG, mabA-inhA, oxyR-ahpC, and rrs. The efficiency of MF-qRT-PCR assay was tested using 261 bacterial isolates and 33 clinical sputum specimens. Among these samples, 227 Mycobacterium tuberculosis isolates were analyzed using drug susceptibility testing (DST), DNA sequencing and MF-qRT-PCR assay. Compared with DST, MF-qRT-PCR sensitivity and specificity for RIF-resistance were 94.6 and 100%, respectively. And the detection sensitivity and specificity for INH-resistance were 85.9 and 95.3%, respectively. Compared with DNA sequencing, the sensitivity and specificity of our assay were 97.2 and 100% for RIF-resistance and 97.9 and 96.4% for INH-resistance. Compared with Phenotypic strain identification, MF-qRT-PCR can distinguish 227 M. tuberculosis complexes (MTC) from 34 Non-tuberculous mycobacteria (NTM) isolates with 100% accuracy rate. MF-qRT-PCR assay was an efficient, accurate, reliable, and easy-operated method for detection of RIF and INH-resistance, and distinction of MTC and NTM of clinical isolates.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
Edited by: Pere-Joan Cardona, Universitat Autònoma de Barcelona, Spain
Reviewed by: Thomas Dandekar, University of Wuerzburg, Germany; Paras Jain, Albert Einstein College of Medicine, USA; Fredy Gamboa, Pontificia Universidad Javeriana, Colombia
This article was submitted to Infectious Diseases, a section of the journal Frontiers in Microbiology
First authors.
ISSN:1664-302X
1664-302X
DOI:10.3389/fmicb.2016.00618