分枝杆菌分型三重PCR检测方法的建立及应用

doi:10.16431/j.cnki.1671-7236.2016.01.002

Abstract

Abstract: This study was aimed to establish a triple PCR method to rapidly identify Mycobacterium species, and evaluate its testing reliability.Three pairs of primer that were respectively specific to rv 3036c, rv 1970f and pncA genes of Mycobacterium were designed to establish a triple PCR for preliminary identification of Mycobacterium tuberculosis(M.tuberculosis), Mycobacterium bovis(M.bovis) and other Mycobacterium spp.PCR products were the expected sizes of 500(rv3036c), 125(rv1970f) and 249 bp(pncA), and contained two DNA bands(500 and 125 bp) with M.tuberculosis DNA template, two DNA bands(500 and 249 bp) with M.bovis DNA template.No band or non-specific band appeared with Mycobacterium spp.except M.tuberculosis and M.bovis DNA templates.The sensitivity of the triple PCR was calculated to 50 pg/μL template of genomic DNA.86 acid-fast bacteria were detected by the triple PCR, 16S rDNA and ITS gene sequencing, growth test and biochemical test, and the results were consistent between triple PCR and 16S rDNA and ITS gene sequencing.The detecting accuracy of triple PCR was 100%, and higher than growth test and biochemical test.

Key words: Mycobacterium; Mycobacterium tuberculosis; Mycobacterium bovis; triple PCR; preliminary identification

CLC Number:

S852.61⁺8

LOU Zhong-zi, LIU Cong-nuan, JIA Wan-zhong, ZHOU Ji-zhang, YAN Hong-bin, CAO Xiao-an, LI Li, LI Zhao-cai, FU Bao-quan. Establishment and Application of Mycobacterium Typing Triple PCR Detection Method[J]. , 2016, 43(1): 8-15.

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Establishment and Application of Mycobacterium Typing Triple PCR Detection Method

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