蛋白酶K预处理在SARS-CoV-2核酸提取中的应用探讨  

Application of protease K pretreatment in nucleic acid extraction of SARS-COV-2

作　　者：王念[1] 巫丽娟[1] 周易[1] 宋兴勃[1] 叶远馨[1] 钟慧钰 周汶静[1] 陶昕彤 刘堂喻亨 宋佳佳 斯艳君 陆小军[1] 应斌武[1] WANG Nian;WU Lijuan;ZHOU Yi;SONG Xingbo;YE Yuanxin;ZHONG Huiyu;ZHOU Wenjing;TAO Xintong;LIU Tangyuheng;SONG Jiajia;SI Yanjun;LU Xiaojun;YING Binwu(Department of Laboratory Medicine,West China Hospital of Sichuan University,Chengdu,Sichuan 610041,China)

机构地区：[1]四川大学华西医院实验医学科,四川成都610041

出　　处：《国际检验医学杂志》2021年第1期30-33,共4页International Journal of Laboratory Medicine

基　　金：四川省科技厅新型冠状病毒科技攻关应急项目(2020YFS004);四川大学nCoV应急项目(2020scunCoV应急10003);四川大学华西医院新型冠状病毒肺炎疫情科技攻关项目(HX-2019-nCoV-008、HX-2019-nCoV-048)。

摘　　要：目的探讨蛋白酶K(PK)在新型冠状病毒(SARS-CoV-2)不同类型标本中的应用价值。方法通过对临床上确诊新型冠状病毒肺炎(COVID-19)患者核酸检测阳性的50份鼻咽拭子标本和20份粪便标本分别进行平行实验,向其中一份加入PK进行预处理(PK组),而另外一份则不加入PK(未处理组)。使用NucliSENS easyMAG磁珠提取系统对标本进行RNA提取,提取的RNA经实时荧光定量PCR(qPCR)进行SARS-CoV-2核酸检测。结果对50份鼻咽拭子标本和20份粪便标本的qPCR结果中ORF基因和N基因的循环阈值进行分析,未处理组和PK组实验结果比较,差异无统计学意义(P>0.05)。结论在使用NucliSENS easyMAG磁珠提取系统对SARS-CoV-2鼻咽拭子标本和粪便标本进行提取时,PK的预处理对核酸检测结果没有影响。Objective To explore the application of protease K(PK)in different types of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Methods Corona virus disease 2019(COVID-19)patients with clinically confirmed 50 cases of nasopharyngeal swabs and 20 stool specimens were tested for parallel treatment.One group was pretreated with PK(group PK),while the other was not treated with PK(untreated group).RNA was extracted by NucliSENS easyMAG magnetic bead extraction system.The extracted RNA was detected by real-time fluorescent quantitative PCR(qPCR).Results The circulating thresholds of ORF gene and N gene in 50 nasopharyngeal swabs and 20 fecal samples were analyzed.There was no significant difference between untreated group and PK group(P>0.05).Conclusion The pretreatment of PK has no effect on the results of nucleic acid detection in the nasopharynx swab and fecal samples of SARS-CoV-2 by NucliSENS easyMAG magnetic bead extraction system.

关 键 词：新型冠状病毒 蛋白酶K 核酸提取 实时荧光定量PCR 

分 类 号：R446.5[医药卫生—诊断学]