摘要
目的从四川省成都地区采集三带喙库蚊样品,快速检测蚊虫中的流行性乙型脑炎病毒（JEV）,并确定JEV基因型别。方法采用实时荧光RT-PCR方法检测蚊体内的JEV。利用一步法RT-PCR试剂盒扩增阳性样品Pr M区段特异性核苷酸序列,测序后应用Clustal X1.83和Mega 5.0软件构建系统进化树,并进行基因分型和同源性分析。结果 2012年采集三带喙库蚊雌蚊10 656只,分为219份,检测出7份JEV阳性核酸样品,阳性率为3.2%。扩增Pr M区域特异性核苷酸序列674 bp,经鉴定均为基因Ⅰ型。与国内外部分GⅠ型的相关序列比较,核苷酸同源性为91.5%~100%,氨基酸同源性为94.9%~100%,其中与四川省SC09-X08株同源性最高。结论实时荧光RT-PCR法能快速检测三带喙库蚊中的JEV,成都地区存在基因Ⅰ型JEV流行。
Objective To rapidly detect Japanese encephalitis virus （JEV） from Culex tritaeniorhynchus in Chengdu by real-time fluorescence reverse-transcriptase polymerase chain reaction （RT-PCR） assay, and to analyze the genotype of JE~. Methods Real-time fluorescence RT-PCR was used for detection of JEV infection. PrM segments of JEV from the nucleonic acid detected were amplified by one step RT-PCR, and were sequenced. Clustal X1.83 and Mega 5.0 were used to construct phylogenetic tree and analyze genotype and homology. Results A total of 10 656 mosquitoes of Cx. tritaeniorhynchus were collected with 219 pools in 2012. Seven pools of Cx. tritaeniorhynchus were positive for JEV at 3.2%. Based on sequence analysis, seven samples were identified to be genotype I JEV with the 674 bp sequence of PrM. Compared with some sequences of JEV G I, the seven samples showed nucleotide and amino acid homologies of 91.5%-100% and 94.9%-100%, respectively, and were highly homology with the Sichuan strain of JEV （SC09-X08）. Conclusion The prevalent JEV from Cx. tritaeniorhynchus in Chengdu was quickly detected by real-time fluorescence RT-PCR assay and confirmed to be genotype I.
出处
《中国媒介生物学及控制杂志》
CAS
CSCD
2015年第2期137-140,共4页
Chinese Journal of Vector Biology and Control
