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当前位置: 首页 > 文章 > 赤羽病病毒套式RT-PCR检测方法的建立与初步应用 畜牧与兽医 2010,42 (6) 1-4
Position: Home > Articles > Developmental and potential application of a nested RT-PCR for the detection of akabane virus Animal Husbandry & Veterinary Medicine 2010,42 (6) 1-4

赤羽病病毒套式RT-PCR检测方法的建立与初步应用

作  者：
张吉红;黄素文;闻伟刚;李如松
单  位：
宁波出入境检验检疫局
关键词：
赤羽病病毒;套式RT-PCR;S基因
摘  要：
根据GenBank中已发表的赤羽病病毒(AKAV)的S基因序列,设计了3条特异性引物,建立了检测AKAV的套式RT-PCR方法。特异性试验表明,该方法可以特异扩增出AKAV的S基因片段,但从牛病毒性腹泻病毒(BVDV)、传染性牛鼻气管炎病毒(IBRV)等13种对照病毒中均不能扩增出目的条带。敏感性试验表明,套式RT-PCR能够扩增10-5稀释度的病毒RNA(核酸含量约1.18 ng/μL),比普通RT-PCR高出1 000倍。用此方法检测130份奶牛血清样品与1份流产胎儿病料,均未检测到阳性样本,但AKAV在血清模拟样品中可被有效检出。研究结果表明,该方法灵敏、特异,为AKAV的检测提供了一个快速、有效的手段。
译  名：
Developmental and potential application of a nested RT-PCR for the detection of akabane virus
作  者：
ZHANG Ji-hong,HUANG Su-wen,WEN Wei-gang,LI Ru-song(Ningbo Entry-Exit Inspection and QuarantineBurean,Ningbo 315012,China)
关键词：
akabane virus(AKAV);nested RT-PCR;S gene
摘  要：
A nested RT-PCR assay for detecting akabane virus(AKAV) was established using 3 primers.The primers were designed based on S gene information of AKAV in GenBank.The nested RT-PCR could only amplify the S gene fragment from AKAV,but not from 13 other viruses such as bovine viral diarrhoea virus(BVDV),infectious bovine rhinotracheitis virus(IBRV),and so on.The lowest detection limit for AKAV RNA was 10-5 dilution(RNA concentration was 1.18 ng/μL).The assay was 1 000 times more sensitive than normal RT-PCR.One hundreds and thirty cow serum samples and one sample from aborted fetus of dairy cow were tested by the nested RT-PCR for the detection of AKAV.The results showed that the above specimens were negative for AKAV.But in the mimic serum specimens,positive results could be obtained.It suggested that the nested RT-PCR was easy,specific and sensitive for detecting AKAV.It could be used for a rapid detection of AKAV.

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