一种基于母血浆的新型冠状病毒核酸检测体系的建立  

Development a novel system for detection RNA of SARS-CoV-2 based on maternal plasma

作　　者：林彭思远 杨旭 周世豪[1] 卜秀芬 刘激扬 贺骏[1] LINPENG Siyuan;YANG Xu;ZHOU Shihao;BU Xiufen;LIU Jiyang;HE Jun(Department of Genetics and Eugenics,Changsha Hospital for Maternal and Child Health Care,Changsha Hu nan 410007,China;Southern Medical University,Guangzhou Guangdong 510515,China,Guangzhou Darui Biotehnology Co.,Ltd.Guangzhou Guangdong 510663,China;The First Hospital of Changsha,Changsha Hu nan 410005,China)

机构地区：[1]长沙市妇幼保健院遗传优生科,湖南长沙410007 [2]南方医科大学,广州市达瑞生物技术股份有限公司,广东广州510515 [3]长沙市第一医院,湖南长沙410005

出　　处：《转化医学杂志》2021年第3期170-174,共5页Translational Medicine Journal

基　　金：湖南省重点领域研发计划—社会发展领域重点研发项目(2020SK3044)。

摘　　要：目的通过改良染色体非整倍体无创产前检测(Non-Invasive Prenatal Testing,NIPT)的文库构建方法,开发一种基于母血浆的新型冠状病毒(SARS-CoV-2)核酸检测方法,实现NIPT与SARS-CoV-2的整合性筛查。方法收集2020年7月1日-2020年9月30日在长沙市妇幼保健院就诊行NIPT检测的孕妇50例,将孕妇血浆等分为两份,一份常规提取游离DNA,另一份提取SARS-CoV-2核酸并逆转录,两种提取物等比例混合后按照NIPT常规流程进行扩增、测序和分析。改良测试文库分为三组,正常孕妇组,三体孕妇组和模拟SARS-CoV-2孕妇组,其中模拟SARS-CoV-2孕妇组又设置三个不同的病毒浓度梯度。结果①正常孕妇组所有样本NIPT检测均为阴性,三体孕妇组4例为21三体、4例为18三体、1例为13三体、1例为21三体合并13三体,均与实际相符。②模拟SARS-CoV-2孕妇组三个病毒浓度梯度均可实现SARS-CoV-2核酸的检测,且NIPT检测结果为阴性,与实际相符。③SARS-CoV-2序列占比与病毒拷贝数的相关系数为0.997,提示该方法有潜力用于SARS-CoV-2的定量或半定量分析。结论本研究开发的NIPT-SARS-CoV-2技术可以在孕妇常规产检过程中,采集一管外周血即达到NIPT检测和孕妇SARS-CoV-2核酸检测的目的,实现孕产妇这一易感人群的SARS-CoV-2全面筛查,保证孕妇和新生儿得到最优化的管理与救治。Objective Through optimizing the noninvasive prenatal testing(NIPT)library,developing a new system for detection RNA of SARS-CoV-2 based on maternal plasma to achieve an integrated screening for fetal aneuploidy and SARS-CoV-2 RNA detecting.Method The maternal plasma was divided into two parts,one was extracting for free DNA,the other was extracting for SARS-CoV-2 RNA and reverse transcription.The two extracts were mixed in equal proportion,and sequencing and analysing were performed according to the NIPT routine process.The improved test library was divided into three groups:normal pregnant women group,trisomy pregnant woman group,and simulated SARS-CoV-2 pregnant women group.There were three different virus concentration gradients in the simulated SARS-CoV-2 pregnant women group.Results①The NIPT results of all ten cases in normal pregnant women group were negative.In trisomy pregnant group,4 cases were trisomy 21,4 cases were trisomy 18,1 case was trisomy 13,and 1 case was trisomy 21 combined with trisomy 13.The NIPT results of all cases in the two groups were consistent with the templates.②SARS-CoV-2 was detected in all cases of three virus concentration gradients of simulated SARS-CoV-2 pregnant women group,and the NIPT results were all negative,which was consistent with the actual situation.③The correlation coefficient between the proportion of SARS-CoV-2 sequences and the copy number of SARS-CoV-2 was 0.997,indicating that this method has the potential for quantitative or semi-quantitative analysis of SARS-CoV-2.Conclusion Through only a tube of peripheral blood of pregnant women in the routine prenatal examination process,the technology of NIPT-SARS-CoV-2 which developed in this study can achieve the purpose of NIPT and SARS-CoV-2 RNA detecting at the same time.

关 键 词：新型冠状病毒 无创产前检测 无症状携带者 孕妇 

分 类 号：R563.1[医药卫生—呼吸系统]