[1]赵国清,尹斐斐,安泓霏,等.犬瘟热与犬副流感病毒双重一步法RT-PCR检测方法的建立[J].中国预防兽医学报,2018,(09):818-821.[doi:0.3969/j.issn.1008-0589.201712001]
 ZHAO Guo-qing,YIN Fei-fei,AN Hong-fei,et al.Establishment of the duplex one-step reverse transcription PCR for detection of canine distemper virus and canine parainfluenza virus[J].Chinese journal of preventive veterinary medicine,2018,(09):818-821.[doi:0.3969/j.issn.1008-0589.201712001]
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犬瘟热与犬副流感病毒双重一步法RT-PCR检测方法的建立()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年09
页码:
818-821
栏目:
诊断和检测技术
出版日期:
2018-09-17

文章信息/Info

Title:
Establishment of the duplex one-step reverse transcription PCR for detection of canine distemper virus and canine parainfluenza virus
文章编号:
1008-0589(2018)9-0818-04
作者:
 

赵国清12尹斐斐4安泓霏1王贵升3*胡永浩1*

 (1. 甘肃农业大学,甘肃 兰州730070;2. 威海市畜牧兽医局,山东 威海 264200;3. 山东省动物疫病预防
与控制中心,山东 济南 250022;4. 青岛市动物疫病预防控制中心,山东 青岛 266071)
Author(s):
 

ZHAO Guo-qing12 YIN Fei-fei4 AN Hong-fei1 WANG Gui-sheng3* HU Yong-hao1*

 (1.Gansu Agricultural University, Lanzhou 730070, China; 2. Weihai Animal Husbandry and Veterinary Bureau, Weihai 264200, China;
3. Shandong Provincical Center for Animal Disease Control and Prevention, Jinan 250022, China;
4. Qingdao Center for Animal Disease Control and Prevention, Qingdao 266071, China)
关键词:
犬瘟热病毒犬副流感病毒RT-PCR检测
Keywords:
 canine distemper virus  canine parainfluenza virus  RT-PCR  detection
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201712001
文献标志码:
A
摘要:
 为建立可以同时检测犬瘟热病毒(CDV)和犬副流感病毒(CPIV)的检测方法,依据 GenBank 中登录的CDV的H基因和CPIV的NP基因保守序列,设计合成扩增序列分别为593 bp (CDV H基因)、1 530 bp (CPIV NP基因)的2对引物,通过优化反应条件,建立一种能够同时检测CDV和CPIV的双重一步法RT-PCR检测方法。特异性和敏感性试验显示,该方法能特异地检测CDV和CPIV,其最低检测限分别为3.58×106 拷贝/μL 和1.74×106 拷贝/μL。对51份临床疑似病料样品进行检测,同时分别采用商品化的CDV和CPIV抗原检测试剂盒进行检测,结果二者符合率为100 %。该结果表明本实验所建立的双重一步法RT-PCR方法的灵敏度及特异性均较好,可用于临床相关疫病的诊断。
Abstract:
To develop a method for simultaneous detection of canine distemper virus (CDV) and canine parainfluenza virus (CPIV), a duplex one-step reverse transcription PCR (RT-PCR) was established with two pairs of specific primers designed according to the H gene of CDV and NP gene of CPIV. Under the optimized conditions of the duplex one-step RT-PCR, two special fragments of CDV (593bp) and CPIV (1,530bp) were amplified from the genomic RNAs of CDV and CPIV. It was showed that the duplex one-step RT-PCR assay was specificity and sensitivity with a detection limit of 3.58×106copies/μL and 1.74×106copies/μL. Fifty-one suspected clinical samples were detected by the duplex one-step RT-PCR assay and commercial CDV and CPIV antigen test kits, respectively. It was showed that the coincidence rate of the two detection methods was 100%. The results showed that the sensitivity and specificity of the duplex one-step RT-PCR method could be application in clinical diagnoses of the related diseases.

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(本文编辑:李  娜)

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更新日期/Last Update: 2018-10-19