HPV16与HPV18(E6/E7基因)RNA假病毒核酸标准物质定值研究

doi:10.3969/j.issn.1000-1158.2023.03.12

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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)×10²copies/μL,(5.9±1.1)×10²copies/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。

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	李会杰
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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

Abstract：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)×10²copies/μL, (5.9±1.1)×10²copies/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.

Key words： metrology human papilloma virus (HPV) nucleic acid detection reference material one-step reverse transcriptase digital PCR technology

收稿日期: 2022-07-18 发布日期: 2023-03-08

PACS:

TB99

基金资助:国家市场监督管理总局科技计划项目(2021MK149)

通讯作者: 陈桂芳(1998-),云南省宣威市,沈阳化工大学硕士研究生,主要研究方向为生物核酸计量。Email:503211735@qq.com 杨佳怡(1989-),甘肃兰州人,中国计量科学研究院前沿计量科学中心副研究员,从事生物核酸计量方面的研究工作。Email:yangjy2018@nim.ac.cn 董莲华(1982-),河北涞水人,中国计量科学研究院前沿计量科学中心研究员,从事生物核酸计量方面的研究工作。Email:donglh@nim.ac.cn E-mail: yangjy2018@nim.ac.cn

作者简介: 李会杰(1994-),河南周口人,深圳中国计量科学研究院技术创新研究院硕士研究生,主要研究方向为生物核酸计量。Email:15713680318@163.com

引用本文:

李会杰,陈桂芳,高运华,杨佳怡,董莲华. HPV16与HPV18(E6/E7基因)RNA假病毒核酸标准物质定值研究[J]. 计量学报, 2023, 44(3): 401-409.
LI Hui-jie,CHEN Gui-fang,GAO Yun-hua,YANG Jia-yi,DONG Lian-hua. The Research of Quantification for HPV16 and HPV18(E6/E7) RNA Pseudovirus Nucleic Acid Reference Materials. Acta Metrologica Sinica, 2023, 44(3): 401-409.

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