化脓隐秘杆菌TaqMan实时荧光定量PCR检测方法的建立

doi:10.16431/j.cnki.1671-7236.2015.05.009

›› 2015, Vol. 42 ›› Issue (5): 1093-1097.doi: 10.16431/j.cnki.1671-7236.2015.05.009

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Establishment of TaqMan Real-time PCR Assay for Detection of Arcanobacterium pyogenes

LIU Er-long¹, LU Li¹, TANG Jie², LV Ying-zi¹, ZHENG Gao-bin¹, JIANG Xiang¹

1. Guangdong Entry-exit Inspection and Quarantine Bureau, Guangzhou 510730, China;
2. Shaanxi Institute of Zoology, Xi'an, Shaanxi 710032, China

Received:2014-11-10 Online:2015-05-20 Published:2015-06-02

Abstract

Abstract: The study was aimed to establish a specific and sensitive TaqMan Real-time PCR assay for detection of Arcanobacterium pyogenes (A.pyogenes).Based on the conservative sequence of pyolysin (PLO) gene of A.pyogenes published in GenBank, specific primers and TaqMan probes were designed. The TaqMan Real-time PCR assay was established, and the specificity and sensitivity were tested.The specificity test results showed that only A.pyogenes exhibited typical curves.The detection sensitivity of this assay was 77.6 fg genomic DNA per 20 μL reaction, and 63 CFU/mL for pure cultures.16 out of 23 clinical samples were positive detected by the TaqMan Real-time PCR assay, which were consistent with API Coryne identification.The TaqMan Real-time PCR assay developed in this study was specific and sensitive for detection of A.pyogenes, and it could be used for identification and epidemiological investigation of A.pyogenes.

Key words: Arcanobacterium pyogenes(A.pyogenes); TaqMan Real-time PCR; pyolysin (PLO)

CLC Number:

S852.61

LIU Er-long, LU Li, TANG Jie, LV Ying-zi, ZHENG Gao-bin, JIANG Xiang. Establishment of TaqMan Real-time PCR Assay for Detection of Arcanobacterium pyogenes[J]. , 2015, 42(5): 1093-1097.

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Establishment of TaqMan Real-time PCR Assay for Detection of Arcanobacterium pyogenes

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