A novel method for diagnosis of smear-negative tuberculosis patients by combining a random unbiased Phi29 amplification with a specific real-time PCR - 16/06/15

Doi : 10.1016/j.tube.2015.03.011

Yu Pang ^a,^{^{1
These authors contributed equally to this paper.}}, Jie Lu ^b,^{^{1
These authors contributed equally to this paper.}}, Jian Yang ^c, Yufeng Wang ^a, Chad Cohen ^d, Xin Ni ^b,^⁎ , Yanlin Zhao ^a,^⁎
^a National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China
^b Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, Beijing, China
^c Shaanxi Tuberculosis Dispensary, Shaanxi, China
^d McGill International TB Centre, Montreal, Quebec, Canada

^∗Corresponding author. National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155, Changbai Road, Changping District, Beijing 102206, China. Tel.: +86 10 5890 0777; fax: +86 10 5890 0779.^∗∗Corresponding author. Beijing Children's Hospital, 56 Nanlishi Road, Xicheng, Beijing 100045, China. Tel.: +86 10 59616688; fax: +86 10 59718700.

Summary

In this study, we develop a novel method for diagnosis of smear-negative tuberculosis patients by performing a random unbiased Phi29 amplification prior to the use of a specific real-time PCR. The limit of detection (LOD) of the conventional real-time PCR was 100 colony-forming units (CFU) of MTB genome/reaction, while the REPLI real-time PCR assay could detect 0.4 CFU/reaction. In comparison with the conventional real-time PCR, REPLI real-time PCR shows better sensitivity for the detection of smear-negative tuberculosis (P = 0.015).

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Keywords : Tuberculosis, Diagnostics, Smear negative, REPLI real-time PCR

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Vol 95 - N° 4

P. 411-414 - juillet 2015 Retour au numéro

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A novel method for diagnosis of smear-negative tuberculosis patients by combining a random unbiased Phi29 amplification with a specific real-time PCR - 16/06/15

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