2012 ; 22(9):
|Author||Li Juan Jiang, Wen Juan Wu, Hai Wu, Son Sik Ryang, Jian Zhou, Wei Wu, Tao Li, Jian Guo, Hong Hai Wang, Shui Hua Lu, Yao Li|
|Affiliation||State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, P.R. China|
|Title||Rapid Detection and Monitoring Therapeutic Efficacy of Mycobacterium tuberculosis Complex Using a Novel Real-Time Assay|
J. Microbiol. Biotechnol.2012 ; 22(9):
|Abstract||We combined real-time RT-PCR and real-time PCR (R/P)
assays using a hydrolysis probe to detect Mycobacterium
tuberculosis complex (MTBC)-specific 16S rRNA and its
rRNA gene (rDNA). The assay was applied to 28 nonrespiratory
and 207 respiratory specimens from 218
patients. Total nucleic acids (including RNA and DNA)
were extracted from samples, and results were considered
positive if the repeat RT-PCR threshold cycle was ≤35 and
the ratio of real-time RT-PCR and real-time PCR load
was ≥1.51. The results were compared with those from
existing methods, including smear, culture, and real-time
PCR. Following resolution of the discrepant results
between R/P assay and culture, the overall sensitivity,
specificity, positive predictive values (PPV), and negative
predictive values (NPV) of all samples (including nonrespiratory
and respiratory specimens) were 98.2%, 97.2%,
91.7%, and 99.4%, respectively, for R/P assay, and 83.9%,
89.9%, 72.3%, and 94.7%, respectively, for real-time
PCR. Furthermore, the R/P assay of four patient samples
showed a higher ratio before treatment than after several
days of treatment. We conclude that the R/P assay is a
rapid and accurate method for direct detection of MTBC,
which can distinguish viable and nonviable MTBC, and
thus may guide patient therapy and public health decisions.|
|Keywords||Mycobacterium tuberculosis complex, TaqMan PCR, therapeutic efficacy monitoring|
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