Absolute quantitative real-time RT-PCR assay for rapid detection of viral hemorrhagic septicemia virus (VHSV) with Taqman MGB probe

被引：0
|
作者：
Xu J. [1 ]
Zhang N. [1 ]
Jiang Y. [1 ]
Zhang L. [2 ]
Xia C. [1 ]
机构：
[1] Key Laboratory of Preventive Veterinary Medicine of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University
[2] Beijing Entry-Exit Inspection and Quarantine Bureau
来源：
Gaojishu Tongxin/Chinese High Technology Letters | 2010年 / 20卷 / 02期
关键词：
Quantitative detection; Real-time RT-PCR; Taqman MGB probe; Viral hemorrhagic septicemia virus (VHSV);
D O I：
10.3772/j.issn.1002-0470.2010.02.018
中图分类号：
学科分类号：
摘要：
An absolute quantitative (AQ) real-time RT-PCR (AQ-RT-PCR) method is established and developed for viral hemorrhagic septicemia virus (VHSV) detection. Firstly, the Taqman MGB probe and the primers are designed from highly conserved regions of nucleoprotein (N) gene of VHSV. Secondly, the AQ-RT-PCR method is established using the quantitative standard samples obtained from the in vitro transcripted VHSV N gene. The comparison of the AQ-RT-PCR with the conventional RT-PCR shows that the AQ-RT-PCR is more specific and there are no cross reactions with other fish Rhabdoviridae viruses. The linear range of the AQ-RT-PCR assay is from 1010 copies/reaction to 100 copies/reaction. The low quantitative detection limit is 100 copies/reaction. The sensitivity of the AQ-RT-PCR is higher than the conventional RT-PCR by five orders of magnitude and also by one order of magnitude than the nested PCR. The AQ-RT-PCR method will be efficiently used in entr-exit detection of VHSV.
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页码：208 / 213
页数：5
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