HPV16与HPV18(E6/E7基因)RNA假病毒核酸标准物质定值研究  

The Research of Quantification for HPV16 and HPV18(E6/E7)RNA Pseudovirus Nucleic Acid Reference Materials

作　　者：李会杰 陈桂芳 高运华[2] 杨佳怡 董莲华[2] LI Hui-jie;CHEN Gui-fang;GAO Yun-hua;YANG Jiayi;DONG Lian-hua(Shenzhen Institute for Technology Innovation,National Institute of Metrology,Shenzhen,Guangdong 518107,China;Center for Advanced Measurement Science,National Institute of Metrology,Beijing 100029,China;College of Chemistry Engineering,Shenyang University of Chemistry Technology,Shenyang,Liaoning 110142,China)

机构地区：[1]深圳中国计量科学研究院技术创新研究院,广东深圳518107 [2]中国计量科学研究院前沿计量科学中心,北京100029 [3]沈阳化工大学化学工程学院,辽宁沈阳110142

出　　处：《计量学报》2023年第3期401-409,共9页Acta Metrologica Sinica

基　　金：国家市场监督管理总局科技计划项目(2021MK149)。

摘　　要：分别以含HPV16和HPV18 E6/E7 RNA序列的假病毒为候选材料,采用一步法逆转录数字PCR方法对E6/E7 RNA进行定量检测,研制了HPV16和HPV18(E6/E7基因)RNA假病毒核酸标准物质(NIM-RM5231和NIM-RM5232)。2种标准物质的均匀性和稳定性良好,标准值及扩展不确定度(k=2)分别为:(9.7±1.8)×10^(2)copies/μL,(5.9±1.1)×10^(2)copies/μL。在多家实验室对2种标准物质进行定值验证,定值结果的相对标准偏差均小于10%。所研制的标准物质可为高危型HPV E6/E7 RNA的相关检测的质量控制提供参考。Pseudoviruses containing E6/E7 RNA sequences of HPV16 and HPV18 were selected as reference materials.One-step reverse transcription digital PCR method was used to quantify content of E6/E7 RNA.HPV16 and HPV18(E6/E7)RNA pseudoviruses were prepared and certificated(NIM-RM5231 and NIM-RM5232),which showed good uniformity and stability.The reference values with expanded uncertainty of the two reference materials were:(9.7±1.8)×10^(2)copies/μL,(5.9±1.1)×10^(2)copies/μL.The values were verified by multiple laboratories,and the relative standard deviations of the determination results were all less than 10%.The reference materials can provide reference for the quality control of the related detection of high-risk HPV E6/E7 RNA.

关 键 词：计量学 人乳头瘤病毒 核酸检测 标准物质 一步逆转录酶 数字PCR技术 

分 类 号：TB99[一般工业技术—计量学]