基于QuanNUA基因分型系统开发的用于SARS-CoV-2变异株分型检测的方法,可同时检测新冠毒株S基因7个突变位点9种突变分型用于新冠病毒及其变异毒株检测。
该方法充分利用核酸质谱技术的多位点检测优势,准确性高,适用于新冠变异株分型检测;同时,该方法可实现样本的高通量快速检测,检测周期短,从PCR扩增到质谱检测,7h内可完成96份样本的检测;而且,该方法灵活性高,当出现新型新冠突变株时,可在现有检测方案中添加特异性探针用于新增突变位点检测。
该方法是为新冠病毒变异株分型提供一种快速、高通量,高准确度的新型检测方法。
使用QuanNUA基因分型系统检测样本的基因分型,该系统由融智生物提供,包括引物设计软件、试剂盒、MALDI-TOF MS和操作软件。检测主要流程包括核酸反应产物制备和MALDI-TOF MS检测。
制备核酸反应产物
表1▲
用于SARS-CoV-2变异株分型检测靶点的质量探针
图2▲
SARS-CoV-2变异株分型检测所用质量探针延伸前(a)、SARS-CoV-2非变异株延伸后(b)、SARS-CoV-2突变质粒(含HV69-70del, K417N, E484K, N501Y, D614G 和 P681H突变位点)延伸后(c)和SARS-CoV-2突变质粒(含L452R, E484Q, 和P681R突变位点)延伸后(d)的质谱图
该方法是为新冠病毒变异株分型提供一种快速、高通量,高准确度的新型检测方法。
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