李幸乐, 李懿, 王若琳, 苏佳, 康锴, 郭大城, 许汴利, 黄学勇. 应用人胚肺成纤维细胞MRC-5培养分离寨卡病毒研究[J]. 疾病监测, 2018, 33(9): 715-717. DOI: 10.3784/j.issn.1003-9961.2018.09.004
引用本文: 李幸乐, 李懿, 王若琳, 苏佳, 康锴, 郭大城, 许汴利, 黄学勇. 应用人胚肺成纤维细胞MRC-5培养分离寨卡病毒研究[J]. 疾病监测, 2018, 33(9): 715-717. DOI: 10.3784/j.issn.1003-9961.2018.09.004
Xingle Li, Yi Li, Ruolin Wang, Jia Su, Kai Kang, Dacheng Guo, Bianli Xu, Xueyong Huang. Isolation and identification of Zika virus in MRC-5 cell line[J]. Disease Surveillance, 2018, 33(9): 715-717. DOI: 10.3784/j.issn.1003-9961.2018.09.004
Citation: Xingle Li, Yi Li, Ruolin Wang, Jia Su, Kai Kang, Dacheng Guo, Bianli Xu, Xueyong Huang. Isolation and identification of Zika virus in MRC-5 cell line[J]. Disease Surveillance, 2018, 33(9): 715-717. DOI: 10.3784/j.issn.1003-9961.2018.09.004

应用人胚肺成纤维细胞MRC-5培养分离寨卡病毒研究

Isolation and identification of Zika virus in MRC-5 cell line

  • 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。

     

    Abstract:
    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.

     

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