新型冠状病毒 B.1.1.7 变异株特异性数字 PCR 定量方法研究

王霞 ¹，郭若晖 ²，董莲华 ³

Abstract

由于世界卫生组织（WHO）将 B.1.1.7（Alpha）列为 “关切的变异株”，因此建立针对该毒株的特异性高灵敏数字 PCR 检测方法具有重要意义。本研究首先依据该变异株刺突蛋白上H69/V70 缺失、P681H、A570D共 3 个特异性突变位点设计引物与探针，明确 3 种检测体系的线性范围与定量限；再选取其余 4 种 “关切的变异株” 完成特异性验证，并对比不同数字 PCR 平台及不同检测方法的定量准确度。结果证实，本研究建立的数字 PCR 方法准确可靠，可应用于 B.1.1.7 变异株的精准定量检测。

Due to B.1.1.7 (Alpha) being listed as a “variant of concern” by the World Health Organization (WHO), it is particularly important to establish a specific and highly sensitive digital PCR method for it. First, this method required designing primers and probes based on the H69/V70 deletion, P681H, and A570D specific mutation sites of the spike protein, and determining the linear range and limit of quantitation for the three methods. Subsequently, the specificity of the method was verified using four other variants of concern, and quantitative accuracy was compared using different digital PCR platforms and detection methods. Finally, the results indicated that the established digital PCR method was accurate and reliable, and can be used for quantitative detection of B.1.1.7 variant strains.