TaqMan荧光定量PCR监测野猪乙型脑炎病毒

doi:10.7668/hbnxb.2015.S1.064

华北农学报 ›› 2015, Vol. 30 ›› Issue (S1): 355-359. doi: 10.7668/hbnxb.2015.S1.064

所属专题： 畜牧；

TaqMan荧光定量PCR监测野猪乙型脑炎病毒

赵珊¹, 林华², 陈世界², 杨苗², 薛昌华², 郝中香¹, 王彬¹, 文彩芳¹, 颜其贵^1,3

1. 四川农业大学动物医学院, 四川成都 611130;
2. 四川出入境检验检疫局, 四川成都 610041;
3. 四川农业大学动物疫病与人类健康四川省重点实验室, 四川成都 611130

收稿日期:2015-09-10 出版日期:2015-12-28
作者简介:赵珊(1992-),女,四川雅安人,在读硕士,主要从事传染病病原分子生物学及野生动物疾病学研究.
基金资助:
教育部长江学者和创新团队发展计划创新团队项目(IRTO848);四川出入境检验检疫局科技项目(SK201403;SK201411)

Detection of Japanese Cephalitis Virus from Wild Boars by TaqMan-based Real Time PCR

ZHAO Shan¹, LIN Hua², CHEN Shi-jie², YANG Miao², XUE Chang-hua², HAO Zhong-xiang¹, WANG Bin¹, WEN Cai-fang¹, YAN Qi-gui^1,3

1. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;
2. Sichuan Enter-Exit Inspection and Quarantine Bureau, Chengdu 610041, China;
3. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, China

Received:2015-09-10 Published:2015-12-28

摘要/Abstract

摘要： 为了更有效地保护野猪资源,评估其作为种质资源的安全性及人类驯养的风险性,采用 TaqMan荧光定量PCR监测野猪乙型脑炎病毒的携带情况.结果显示,TaqMan荧光定量PCR敏感性是常规PCR检测方法的100倍,检测时间缩短了1/2.重复性试验中,批内及批间变异系数均低于2%.对40份家养野猪血液样品进行检测,TaqMan荧光定量PCR的阳性检出率为10%,国标(GB/T 22333-2008)中RT-PCR的阳性检出率为5%,灵敏度显著提高.表明该方法可用于野猪乙型脑炎的实时监测和流行病学调查,同时也为野猪群乙型脑炎的净化奠定基础.

关键词: 野猪, 乙型脑炎病毒, TaqMan荧光定量PCR

Abstract: The Japanese cephalitis virus of domesticated boars was detected by the method of TaqMan-based Real time PCR in order to protect wild boars more effectively and assess its security as germplasm resources and risk as domestication.The results showed that the sensitivity of TaqMan-based Real time PCR was 100 times than PCR and the test time reduced by almost 1/2.The coefficient variation(CV) of intra/inter-assay for the same sample was less than 2%.Further detection of the 40 blood samples of domesticated boars,the positive rate of TaqMan-based Real time PCR was 10%,more sensitive than RT-PCR which it was 5%.The results proved that the method could support Japanese cephalitis detection and molecular epidemiological investigation,which also could lay the foundation for eradicating Japanese cephalitis from wild boars.

Key words: Wild boars, Japanese encephalitis virus, TaqMan-based Real-time PCR

中图分类号:

赵珊, 林华, 陈世界, 杨苗, 薛昌华, 郝中香, 王彬, 文彩芳, 颜其贵. TaqMan荧光定量PCR监测野猪乙型脑炎病毒[J]. 华北农学报, 2015, 30(S1): 355-359. doi: 10.7668/hbnxb.2015.S1.064.

ZHAO Shan, LIN Hua, CHEN Shi-jie, YANG Miao, XUE Chang-hua, HAO Zhong-xiang, WANG Bin, WEN Cai-fang, YAN Qi-gui. Detection of Japanese Cephalitis Virus from Wild Boars by TaqMan-based Real Time PCR[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(S1): 355-359. doi: 10.7668/hbnxb.2015.S1.064.

http://www.hbnxb.net/CN/Y2015/V30/IS1/355

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