寨卡病毒和基孔肯雅病毒双重检测方法的初步建立

Establishment of a Dual Detection Method for Zika Virus and Chikungunya Virus

本研究初步建立了寨卡病毒(Zika virus,ZIKV)和基孔肯雅病毒(Chikungunya virus,CHIKV)双重PCR检测方法,增加了寨卡病毒和基孔肯雅病毒核酸检测手段。利用寨卡病毒和基孔肯雅病毒NS3基因设计2组特异性扩增引物,并利用合成的NS3质粒验证引物,同时以寨卡病毒和基孔肯雅病毒cDNA为模板评价该方法的特异性和灵敏度。扩增寨卡病毒和基孔肯雅病毒cDNA,分别获得164 bp和230 bp的NS3基因片段,相应对照病毒均未发现164 bp或230 bp的扩增条带,提示寨卡病毒或基孔肯雅病毒NS3扩增序列与相应对照病毒无交叉反应,并且2种病毒双重检测的检测限均为120pg/μL。本研究建立的寨卡病毒和基孔肯雅病毒核酸双重检测方法虽然灵敏性较低,但特异性较好,可作为检测方法满足口岸寨卡病毒和基孔肯雅病毒核酸检测的需要。

This study established a dual PCR method for the detection of Zika virus(ZIKV)and Chikungunya virus(CHIKV),so as to increase the detection methods of ZIKV and CHIKV nucleic acid.ZIKV and CHIKV NS3 genes were selected as targets,and two groups of characteristic primers were designed,and synthetic NS3 plasmids were used to verify the primers.At the same time,ZIKV and CHIKV cDNA were used to evaluate the specificity and sensitivity of the method.The NS3 gene fragments of 164 bp and 230 bp were obtained by using ZIKV and CHIKV cDNA as templates,respectively.No amplification bands was found in the corresponding control viruses,suggesting that there was no cross reaction between NS3 amplification sequences of ZIKV or CHIKV and the corresponding control viruses,and the detection limit of both viruses was 120 pg/μL.The dual detection method of ZIKV and CHIKV nucleic acid established in this study lack ssensitivity,but has good specificity,which can be used as a detection method to meet the needs of ZIKV and CHIKV nucleic acid detection at ports.