用RT-PCR法快速检测鲤春病毒血症病毒基因
DETECTION OF SPRING VIREMIA OF CARP VIRUS)SVCV)GENE USING REVERSE-POLYMERASE CHAIN-REACTION)RT-PCR)
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摘要: 用逆转录多聚酶链式反应(RT-PCR)方法快速、灵敏、特异地检测鲤春病毒血症病毒(SVCV),根据SVC病毒糖蛋白基因序列设计的引物经过RT-PCR和半嵌套PCR(semi-nested-PCR)可扩增出SVC病毒核酸中的714bp和606bp片段.与其他弹状病毒IHNV、VHSV、PFRV没有交叉,具有特异性.灵敏度检测,表明不小于1000个病毒就可检测出阳性结果.本文还对复性温度,引物,Mg2+,Tag酶以及反转录酶的浓度对检测结果的影响进行了探讨.Abstract: Spring viremia of carp virus(SVCV)is a rhabdovirus of fish that causing serious mortalities of carp. A rapid, sensitive and specific detection method was being set up for diagnosis of SVCV used RT PCR and semi nested PCR by the authors. The target regions of the SVCV glycoprotein gene were amplified using R2-F1 and R2-F1 primers. The products were 714bp segment in RT-PCR and 606bp segment in semi nested PCR, respectively. These primers could not amplified RNA of IHNV, VHSV and PFRV. The sensitivity has a detection limit of 1000 TCID50.
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