[Analysis and evaluation of phage amplified biologically assay, DNA sequencing analysis and single-strand conformational polymorphism for the drug susceptibility testing of Mycobacterium tuberculosis].
OBJECTIVE
To rapidly identify drug-resistant Mycobacterium tuberculosis using phenotypic and genotypic methods and to evaluate the clinical significance of rapid phenotypic susceptibility test by phage amplified biologically assay (PhaB).


METHODS
PhaB, DNA sequencing and polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) were performed on the 91 rifampicin (RFP)-resistant strains (including 82 multi-drug resistant Mycobacterium tuberculosis strains), 42 RFP-susceptible strains, 75 ofloxacin (OFLX)-resistant strains and 40 OFLX-susceptible strains at the same time.


RESULTS
The results obtained using PhaB assay, DNA sequencing and PCR-SSCP were compared with the absolute concentration method. For RFP susceptibility, the accordance, sensitivity and specificity of PhaB were 93%, 92% and 95% respectively; the accordance, sensitivity and specificity of DNA sequencing were 93%, 90% and 100% respectively; those of PCR-SSCP were 90%, 86% and 100% respectively. For OFLX susceptibility, the accordance, sensitivity and specificity of PhaB assay were 95%, 95% and 95%; those of DNA sequencing were 80%, 71% and 98% respectively; those of PCR-SSCP were 75%, 63% and 98% respectively.


CONCLUSIONS
PhaB assay is a low-cost, rapid, and sensitive method and shows high accordance with absolute concentration technology. It can give drug susceptibility test results within 48 - 96 h, and is a promising technology in clinical laboratory.
Han
Xi-Qin
XQ
Department of Basic Research, Beijing Tuberculosis and Thoracic Tumor Research Insistute, Beijing 101149, China.
Ma
Yu
Y
Gao
Wei-Wei
WW
Li
Chuan-You
CY
Chen
Xiao-You
XY
Zhang
Zong-de
ZD
Gu
Shu-Xiang
SX
Jiang
Guang-Lu
GL
D000904:Antibiotics, Antitubercular / Q000494:pharmacology; D001483:Base Sequence; D015169:Colony Count, Microbial; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D008826:Microbial Sensitivity Tests / Q000379:methods; D009160:Mycobacteriophages; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods; D018807:Polymorphism, Single-Stranded Conformational; D012293:Rifampin / Q000494:pharmacology*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D000904:Antibiotics, Antitubercular; D012293:Rifampin