摘要
目的应用人胚肺成纤维细胞MRC-5培养并分离鉴定寨卡病毒。方法收集寨卡病毒病确诊病例的精液样本并在MRC-5细胞中进行培养,观察致细胞病变效应(CPE)。采用实时荧光PCR(real-time PCR)方法鉴定培养,同时对培养物进行序列测定和比对分析。结果病例精液样本经MRC-5细胞盲传3代,未观察到明显的特异性致CPE,MRC-5细胞3代培养物寨卡病毒经real-time PCR检测结果呈阳性。毒株命名为Henan/001/2016,GenBank编号为MF593625,属于亚洲基因簇系。Henan/001/2016与Natal RGN株的亲缘关系最为接近,在E基因区段的核苷酸和氨基酸序列同源性分别为99.7%和100%。结论 MRC-5细胞适用于寨卡病毒的分离、培养和鉴定。
Objective To culture, isolate and identify Zika virus in MRC-5 cell line. Methods MRC-5 cell line was used for Zika virus isolation from seminal fluid sample collected from a laboratory conformed Zika virus infection case, the cytopathic effect was observed, and the isolate was identified with real-time PCR. The Zika virus strain was sequenced and evolutionary analysis was conducted. Results The seminal fluid sample was cultured in MRC-5 cell line for three generations in blind passage. No obvious cytopathic effect was observed, but the real-time PCR result was positive. The Zika virus strain belonged to Asian genotype, which was named as Henan/001/2016（GenBank No. MF593625）. The strain was most close to Natal RGN strain genetically, the nucleotide homology and amino acid homology in E gene segment were 99.7% and 100%respectively. Conclusion MRC-5 cell line is suitable for the isolation and identification of Zika virus.
作者
李幸乐
李懿
王若琳
苏佳
康锴
郭大城
许汴利
黄学勇
Li Xingle;Li Yi;Wang Ruolin;Su Jia;Kang Kai;Guo Dacheng;Xu Bianli;Huang Xueyong(Henan Provincial Center for Disease Control and Prevention,Zhengzhou 450016,Henan,China)
出处
《疾病监测》
CAS
2018年第9期715-717,共3页
Disease Surveillance
基金
国家自然科学基金(No.81573204)
河南省科技创新人才基金(No.16410051008)~~
