本文選題：多重PCR + 山羊痘病毒��； 參考：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：山羊痘病毒、綿羊痘病毒、羊口瘡病毒和小反芻獸疫病毒均可單一感染山羊和綿羊,或多重感染。為快速檢測(cè)并鑒別四種病毒引起的疾病,建立多重PCR檢測(cè)方法,并對(duì)建立方法進(jìn)行了實(shí)驗(yàn)條件的優(yōu)化和篩選。根據(jù)GenBank數(shù)據(jù)庫(kù)收錄的山羊痘病毒、綿羊痘病毒、羊口瘡病毒和小反芻獸疫病毒的基因序列,用Primer Primer 5.0設(shè)計(jì)四對(duì)引物用于擴(kuò)增四種病毒,其中山羊痘病毒擴(kuò)增產(chǎn)物長(zhǎng)度152bp,綿羊痘病毒擴(kuò)增產(chǎn)物長(zhǎng)度246bp,羊口瘡病毒擴(kuò)增產(chǎn)物長(zhǎng)度355bp,小反芻獸疫病毒擴(kuò)增產(chǎn)物長(zhǎng)度538bp,優(yōu)化后的最佳試驗(yàn)條件為：變性溫度94℃C,退火溫度53℃,循環(huán)次數(shù)為35次。所建立PCR方法對(duì)牛病毒性腹瀉-黏膜病病毒、豬瘟病毒和牛傳染性鼻氣管炎病毒進(jìn)行擴(kuò)增均呈陰性,說(shuō)明該試驗(yàn)有較好的特異性；該P(yáng)CR反應(yīng)體系可檢測(cè)出山羊痘病毒、綿羊痘病毒、羊口瘡病毒和小反芻獸疫病毒的最小量分別是0.05ng、8.36ng、0.8ng、1.3ng。結(jié)果表明本試驗(yàn)具有快速、特異性強(qiáng)的特點(diǎn),可用于臨床鑒別診斷山羊痘、綿羊痘、羊口瘡和小反芻獸疫。
[Abstract]:Goat pox virus, sheep mouth sore virus and small ruminant zoonotic virus can be single infected goat and sheep, or multiple infections. In order to quickly detect and identify the diseases caused by four viruses, a multiplex PCR method was established, and the experimental conditions were optimized and screened. According to the gene sequence of goat pox virus, sheep mouth sore virus and ruminant virus collected in GenBank database, four pairs of primers were designed with Primer Primer 5.0 to amplify the four viruses. The length of goat pox virus amplification product is 152 BP, sheep pox virus amplification product length is 246 BP, sheep mouth sore virus amplification product length is 355 BP, small ruminant virus amplification product length is 538 BP, the optimized test conditions are as follows: denaturation temperature 94 鈩，

本文編號(hào)：2071500