為實(shí)現(xiàn)袋鼠臨床樣本中皰疹病毒1型（Macropodid herpesvirus 1，MaHV-1）的出入境快速檢測(cè)，基于MaHV-1的gB基因序列設(shè)計(jì)特異引物和TaqMan探針，建立了一種MaHV-1熒光PCR檢測(cè)方法。試驗(yàn)結(jié)果顯示，該方法只對(duì)MaHV-1 gB基因呈現(xiàn)特異性擴(kuò)增，與禽傳染性喉氣管炎病毒、偽狂犬病毒和牛傳染性鼻氣管炎病毒不發(fā)生交叉反應(yīng)，對(duì)陽(yáng)性標(biāo)準(zhǔn)質(zhì)粒對(duì)照（pCR-MaHV-1-gB）的最低檢測(cè)限為8個(gè)拷貝數(shù)/反應(yīng)。該方法的組內(nèi)和組間試驗(yàn)的Ct值變異系數(shù)介于0.17%~0.96%之間，具有良好的重現(xiàn)性。試驗(yàn)結(jié)果表明，本研究建立的實(shí)時(shí)熒光PCR方法可用于袋鼠MaHV-1的病原學(xué)檢測(cè)。
Development of a Real-time PCR for Detection ofMacropodid Herpesvirus 1
For rapidly detecting macropodid herpesvirus 1（MaHV-1）in clinic samples from kangaroos at entry-exit ports，a real-time PCR was developed by designingprimers and probes based on the sequence of gB gene. The results showed themethod could only amplify the MaHV-1 gB gene and no cross-reaction with otherkinds of virus was observed，including infectious bovine bronchitis virus，avian laryngotracheitis virus andpseudorabies virus. The detection limit was 8 copies/reaction for the positivestandard plasmid control（pCR-MaHV-1-gB）. The coefficients of variation（CV）of intra-assay and inter-assay were between 0.17 % to 0.96 %，showing good repeatability. In conclusion，the established method was applicable forpathogenic detection of MaHV-1 from the kangaroos.
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