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H3N2亚型犬流感病毒RT-PCR检测方法的建立

发表时间:2016-11-15  浏览量:1241  下载量:330
全部作者: 王晨曦,王 倩*,胡君宜,孙怡朋
作者单位: 中国农业大学动物医学院
摘 要: 为加强对H3N2亚型犬流感病毒在犬群中流行情况的监测,设计了一对针对H3N2亚型犬流感病毒血凝素(hemagglutinin,HA)基因序列保守区域的引物,并建立了能够鉴别诊断H3N2亚型犬流感病毒的反转录PCR(reverse-transcriptase PCR,RT-PCR)方法。经过验证,建立的方法特异性好,仅能够特异性地检测出H3N2亚型犬流感病毒,与其他感染犬的流感病毒无交叉反应;该方法灵敏度高,最低检出量为1×101 TCID50/100 μL. 为评估其临床应用的潜能,分别用建立的RT-PCR检测方法和传统的病毒分离方法检测了420份犬的鼻咽拭子,二者的阳性检出率完全一致。因此,本方法可作为H3N2亚型犬流感病毒的临床诊断和流行病学调查快速实用的检测工具。
关 键 词: 家畜病毒学;犬流感病毒;H3N2;反转录PCR
Title: Development of RT-PCR assay for detection of avian-origin H3N2 canine influenza viruses
Author: WANG Chenxi, WANG Qian*, HU Junyi, SUN Yipeng
Organization: College of Veterinary Medicine, China Agricultural University
Abstract: In order to strengthen the monitoring of avian-origin H3N2 canine influenza viruses, a pair of specific primers were designed according to the hemagglutinin (HA) genes conserved region of avian-origin H3N2 canine influenza viruses, and a reverse-transcriptase PCR (RT-PCR) assay was developed. The high specificity of this assay showed that there was no cross-reaction with other subtypes canine influenza viruses. This RT-PCR assay was also shown to have high sensitivity and the detection limit was 1×101 TCID50/100 μL. To assess the application of this assay, 420 clinical swabs of dogs were tested using the RT-PCR assay and the conventional virus isolation assay, and the positive rates were completely consistent. Therefore, this RT-PCR assay can serve as a rapid useful tool for clinical diagnosis and epidemiological investigation of avian-origin H3N2 canine influenza viruses.
Key words: livestock virology; canine influenza virus; H3N2; reverse-transcriptase PCR
发表期数: 2016年11月第21期
引用格式: 王晨曦,王 倩*,胡君宜,等. H3N2亚型犬流感病毒RT-PCR检测方法的建立[J]. 中国科技论文在线精品论文,2016,9(21):2260-2264.
 
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