Establishment of a real-time fluorogenic quantitative PCR method for Asian Zika virus detection

Abstract

Objective: To analyze the characteristics of Asian Zika virus (ZIKV) genome sequence and to establish a realtime fluorogenic quantitative PCR method for ZIKV detection.

Methods: The whole genome sequence of Asian ZIKV was analyzed and the conserved region of Asian ZIKV was used to design primers and TaqMan probes. A standard curve was established and its specificity and sensitivity were assessed.

Results: Detection of designed PCR primers and probes were of good specificity, with a lower detection limit of 3.415 × 10 ⁰ copies/μL. There were no cross reactions between African ZIKV, hepatitis C virus and type 1 – 4 dengue virus.

Conclusion: A fluorescence quantitative PCR technique for the detection of Asian Zika virus was established, which could be used in etiologic typing, early diagnosis and prevention of Asian Zika virus.

Abstract

［摘 要］ 目的 分析亚洲型寨卡病毒基因组序列特征, 建立亚洲型寨卡病毒的荧光定量 PCR 检测技术。 方法 通过分析亚洲型寨卡病毒全基因组核酸序列, 选取亚洲型寨卡病毒的保守区设计引物和 TaqMan 探针, 建立标准曲线并检测其特异性与敏感性。 结果 荧光定量 PCR 测试结果表明所设计的引物和探针特异性良好, 对非洲型寨卡病毒、I～Ⅳ型登革病毒和丙型肝炎病毒核酸检测无交叉反应; 同时灵敏度较高, 可达 3.415 × 10 ⁰ copies/μL; 构建了标准曲线, 回归方程为 Y = – 3.227 3logX + 38.79, 相关系数 R2 = 1, 扩增效率 E = 102 %。 结论 建立了一种检测亚洲型寨卡病毒的荧光定量 PCR 技术, 可用于临床亚洲型寨卡病毒感染患者的病原分型、早期诊断及预防。