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 71 4bp和 60 6bp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 1 0 0 0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。
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 1 000 TCID 50 .
出处
《水生生物学报》
CAS
CSCD
北大核心
2002年第5期452-456,共5页
Acta Hydrobiologica Sinica
基金
国家出入境检验检疫局科研项目 (K0 49 1 999)
淡水生态与生物技术国家重点实验室开放基金资助项目 (2 0 0 1PB1 2 )
