Development of TaqMan-qPCR for Detection of Helicobacter muridarum

doi:10.3969/j.issn.1674-5817.2017.05.007

Abstract

Abstract: Objective To establish the sensitive and specific TaqMan-qPCR method for detection of Helicobacter muridarum. Method The specific primers were designed according to the Helicobacter muridarum gyrase B(GyrB) gene, specific fragment was amplified by PCR, and cloned into pMD-19T vector to construct the recombinant plasmid GyrB-19T, which was used as standard DNA of this qPCR method. The qPCR system was optimized and the sensitivity, specificity, repeatability and quantitative range of this method were evaluated. Result The quantitative standard curve from 2×10⁸copies/well to 2×10²copies/well of serial diluted plasmid DNAs showed that they had good liner correlation, the slope of the standard curve was -3.57, R²>0.996, and the lowest detection limit reached 2×10²copies/well. Conclusion This qPCR method showed high sensitivity, specificity and stability, which will be utilized for qualitative and quantitative detection of Helicobacter muridarum.

Key words: Helicobacter muridarum, TaqMan-qPCR

CLC Number:

S855.3Q95-33

WU Miao-li, ZHU Yu-jun, RAO Dan, YUAN Wen, WANG Jing, CONG Feng, HUANG Ren, CHEN Mei-li, GUO Peng-ju. Development of TaqMan-qPCR for Detection of Helicobacter muridarum[J]. Laboratory Animal and Comparative Medicine, 2017, 37(5): 378-382.

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