亚洲型寨卡病毒实时荧光定量PCR检测方法建立

彭淑莹; 覃直然; 陈嘉雯; 余健海; 陆维智; 何晓恩; 朱利; 肖维威; 赵卫

doi:10.11847/zgggws1119617

亚洲型寨卡病毒实时荧光定量PCR检测方法建立

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

摘要

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 × 10⁰ 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。

Abstract:
Objective To analyze the characteristics of Asian Zika virus (ZIKV) genome sequence and to establish a real-time 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 × 10⁰ 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.

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