真鲷虹彩病毒聚合酶链式反应检测方法的建立

Establishment of polymerase chain reaction detection method for red sea bream iridovirus

目的:真鲷虹彩病毒一直都被列入在世界卫生组织(OIE)水生动物疫病病原的名录中,故需要对其检测方法进行深入研究,建立操作简单,普遍适用的检测技术手段。方法:本文针对RSIV片段设计了一对特异性引物,对反应体系和反应程序进行筛选和优化,建立了检测RSIV聚合酶链式反应的方法。并且用优化好的反应体系和反应程序对重组质粒p MD18-T-RSIV进行扩增。结果:研究发现该方法特异性强,与IHNV、IPNV、SVCV和VHSV无交叉反应,并且灵敏度高,可以达到0.2pg。结论:成功的建立了真鲷虹彩病毒常规PCR检测方法,可以为真鲷虹彩病毒病的诊断提供理论依据。

Objective：Red sea bream iridovirus had been included in aquatic animal disease pathogens list by World Health Organization（OIE）,so it is necessary to need a depth study of its detection methods,establish a simple, universally applicable detection technique.Methods： One pair of specific primers was designed based on RSIV fragment,and the reaction system and procedures for screening and optimized to establish a PCR assay for detection of RSIV.and with the optimized reaction system and procedures for the recombinant plasmid pMD18-T- RSIV amplification.Results.the study showed that the method was specificity, had no cross-reactivity with IHNV, IPNV,SVCV and VHSV,and with high sensitivity,it could achieve 0.2pg.Conclusion.Successfully established a red sea bream iridovirus conventional PCR detection methods,and provide a theoretical basis for the diagnosis of red sea bream iridovirus disease.