BMD-PCR奥密克戎变异株核酸快速检测方法的建立  

Establishment of a rapid detection method for SARS-CoV-2 Omicron variant nucleic acid based on BMD-PCR technology

作　　者：焦洋[1] 韩桃利 赵剑虹[1] 张士尧[1] 吕志 王绍成 孙灵利[1] Jiao Yang;Han Taoli;Zhao Jianhong;Zhang Shiyao;Lyu Zhi;Wang Shaocheng;Sun Lingli(Beijing Chaoyang District Center for Disease Control and Prevention,Beijing 100021,China;Liaoning Runji Biotechnology Company,Shenyang 110027,China)

机构地区：[1]北京市朝阳区疾病预防控制中心,北京100021 [2]辽宁润基生物科技有限公司,沈阳110027

出　　处：《中华实验和临床病毒学杂志》2025年第1期109-114,共6页Chinese Journal of Experimental and Clinical Virology

基　　金：北京市朝阳区科技计划课题(CYSF2206);国家重点研发计划项目(2022YFE0102400)。

摘　　要：目的新型冠状病毒(SARS-CoV-2)自然突变率较高,动态监测病毒变异仍是目前新冠防控工作重点。方法本研究基于bubble mediated nucleic acid denaturation(BMD)-PCR技术建立了一种灵敏、快速的SARS-CoV-2奥密克戎变异株及其流行分支JN.1的核酸检测方法。结果该方法具有良好的特异性,与常见的11种经呼吸道和肠道传播的病毒无交叉反应,最低检出限为555 copies/ml。本方法对奥密克戎变异株及其流行分支JN.1的检测结果得到了全基因测序结果的验证,SARS-CoV-2原始核酸检测结果Ct值≤32的50份奥密克戎突变株样本,49份BMD-PCR方法N679K突变位点阳性,结果符合率达到98.00%;30份JN.1分支样本,29份BMD-PCR方法K356T突变位点阳性,结果符合率达到96.67%;SARS-CoV-2原始核酸检测结果35≤Ct值<32的10份样本,7份BMD-PCR方法N679K突变位点阳性,检出率为70%;SARS-CoV-2原始核酸检测结果Ct值>35的10份样本,2份BMD-PCR方法N679K突变位点阳性,检出率为20%。结论该方法能作为高通量的补充方法,用于SARS-CoV-2变异株基因型的初步鉴定。ObjectiveThe SARS-CoV-2 has a high natural mutation rate,and dynamic monitoring of virus variants remains a key focus in current COVID-19 prevention and control efforts.MethodsIn this study,a sensitive and rapid method for detecting SARS-CoV-2 omicron variant nucleic acid was established based on the BMD-PCR technology.ResultsThis method showed good specificity,and had no cross-reactivity with 11 common viruses transmitted via the respiratory and gastrointestinal tracts,and the limit of detection is 555 copies/ml.Compared with SARS-CoV-2 whole-genome sequencing result,among 50 samples with original Ct values≤32 tested for the Omicron variant,49 samples tested positive for the N679K mutation site using BMD-PCR Omicron variant detection,achieving a concordance rate of 98.00%.For 30 samples JN.1 lineage,29 samples tested positive for the K356T mutation site using BMD-PCR JN.1 lineage detection,with a concordance rate of 96.67%.For 10 samples with original SARS-CoV-2 detection Ct values between 35 and 32,7 samples tested positive for the N679K mutation site using BMD-PCR Omicron variant detection,Resultsing in a detection rate of 70.00%.For samples with SARS-CoV-2 nucleic acid detection Ct values>35,the detection rate for the N679K mutation site in the BMD-PCR Omicron variant was 20.00%.ConclusionsThis method can serve as a high-throughput supplementary approach for the preliminary identification of SARS-CoV-2 variant genotypes.

关 键 词：新型冠状病毒 密克戎变异株 快速检测 BMD-PCR 

分 类 号：S85[农业科学—兽医学]