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Position: Home > Articles > Establishment of polymerase chain reaction detection method for red sea bream iridovirus Science and Technology of Food Industry 2014,35 (23) 298-301+307

真鲷虹彩病毒聚合酶链式反应检测方法的建立

作  者:
万雪;赵前程;吴斌;肇慧君;李叶
单  位:
大连海洋大学;辽宁出入境检验检疫局
关键词:
真鲷虹彩病毒;聚合酶链式反应;优化
摘  要:
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。
译  名:
Establishment of polymerase chain reaction detection method for red sea bream iridovirus
作  者:
WAN Xue;ZHAO Qian-cheng;WU Bin;ZHAO Hui-jun;LI Ye;Dalian Ocean University;Liaoning Entry- Exit Inspection and Quarantine Bureau;Liaoning Entry-Exit Inspection and Quarantine Bureau;
关键词:
red sea bream iridovirus;;polymerase chain reaction;;optimization
摘  要:
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 p MD18- TRSIV 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.

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