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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：1612 下载量：444
| 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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