人乳头瘤病毒6型和11型通用型含内源性内参的重组酶 介导的等温扩增技术检测方法的建立

doi:10.19871/j.cnki.xfcrbzz.2022.02.006

新发传染病电子杂志 ›› 2022, Vol. 7 ›› Issue (2): 25-29.doi: 10.19871/j.cnki.xfcrbzz.2022.02.006

人乳头瘤病毒6型和11型通用型含内源性内参的重组酶介导的等温扩增技术检测方法的建立

何安娜¹, 张梦怡¹, 李逢钰⁴, 马学军², 申辛欣², 戎秀格³

1.华北理工大学临床医学院,河北唐山 063000;
2.中国疾病预防控制中心病毒病预防控制所国家卫生健康委员会医学病毒和病毒病重点实验室,北京 102206;
3.唐山工人医院检验科,河北唐山 063000;
4.河北省人民医院检验科,石家庄 050051

收稿日期:2022-01-19 出版日期:2022-05-31 发布日期:2022-07-07
通讯作者: 申辛欣,Email：x616815@126.com
基金资助:
国家重点研发计划（2021YFC2301100）

Establishment of a universal endogenous internally controlled recombinase-aided amplification detection method for human papilloma virus types 6 and 11

He Anna¹, Zhang Mengyi¹, Li Fengyu⁴, Ma Xuejun², Shen Xinxin², Rong Xiuge³

1. College of Clinical Medicine, North China University of Science and Technology,Hebei Tangshan 063210, China;
2. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;
3. Tangshan Gongren Hospital, Hebei Tangshan 063000, China;
4. Laboratory Department, Hebei General Hospital, Shijiazhuang 050051, China

Received:2022-01-19 Online:2022-05-31 Published:2022-07-07

摘要/Abstract

摘要： 目的建立一种通用型含内源性内参的重组酶介导的等温扩增技术（EIC-RAA) 检测人乳头瘤病毒6型（HPV-6）和11型（HPV-11）的方法,以实现HPV-6和HPV-11的简单、准确、快速筛查。方法针对HPV-6和HPV-11的L1片段保守区域设计通用型引物和探针并结合核糖核酸酶P（RNaseP）基因引物和探针作为内参建立了检测HPV-6和HPV-11的EIC-RAA方法;用系列浓度稀释的质粒和不同亚型HPV样本核酸检验其灵敏度、重复性和特异度;收集的230份临床样本进行EIC-RAA和商业化HPV PCR-荧光探针法检测,并对结果进行比较。结果建立的EIC-RAA方法可在39℃条件下30min内实现HPV-6和HPV-11的检测,其灵敏度为10copies/reaction,重复性好,特异度高,EIC-RAA检测结果与商业化HPV PCR-荧光探针法检测结果的符合率为100%。结论建立了一种检测HPV-6和HPV-11的EIC-RAA方法,该检测方法具有简单、准确、快速的优点,在HPV-6和HPV-11的筛查中具有潜在的应用价值。

关键词: 人乳头瘤病毒, 核糖核酸酶P, 内源性内参, 重组酶介导的等温扩增技术

Abstract: Objective To establish a universal endogenous internally controlled recombinase-aided amplification (EIC-RAA) for the detection of HPV-6 and HPV-11 method, in order to achieve simple, accurate and rapid screening of HPV-6 and HPV-11. Method The EIC-RAA method for detecting HPV-6 and HPV-11 was established by designing universal primers and probes for L1 fragment of the conserved region of HPV-6 and HPV-11, and combining RNaseP gene primers and probes as internal reference. The sensitivity, repeatability and specificity of EIC-RAA method were tested by series of concentration diluted plasmids and nucleic acids from different subtypes of HPV sample. A total of 230 clinical samples were collected and compared for EIC-RAA and commercial HPV PCR-fluorescent probe method. Result The established EIC-RAA method can detect HPV-6 and HPV-11 within 30 minutes at 39℃, with a sensitivity of 10 copies/reaction, good repeatability and high specificity. The coincidence rate between EIC-RAA and commercial HPV PCR-fluorescent probe method was 100%. Conclusion The EIC-RAA showed the advantages of simplicity, accuracy and rapidity in detecting HPV-6 and HPV-11, and has great potential application value in the field screening of HPV-6 and HPV-11.

Key words: Human papilloma virus, Ribonuclease P, Endogenous internally control, Recombinase-assisted amplification

何安娜, 张梦怡, 李逢钰, 马学军, 申辛欣, 戎秀格. 人乳头瘤病毒6型和11型通用型含内源性内参的重组酶介导的等温扩增技术检测方法的建立[J]. 新发传染病电子杂志, 2022, 7(2): 25-29.

He Anna, Zhang Mengyi, Li Fengyu, Ma Xuejun, Shen Xinxin, Rong Xiuge. Establishment of a universal endogenous internally controlled recombinase-aided amplification detection method for human papilloma virus types 6 and 11[J]. Electronic Journal of Emerging Infectious Diseases, 2022, 7(2): 25-29.

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人乳头瘤病毒6型和11型通用型含内源性内参的重组酶介导的等温扩增技术检测方法的建立

Establishment of a universal endogenous internally controlled recombinase-aided amplification detection method for human papilloma virus types 6 and 11

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人乳头瘤病毒6型和11型通用型含内源性内参的重组酶 介导的等温扩增技术检测方法的建立

Establishment of a universal endogenous internally controlled recombinase-aided amplification detection method for human papilloma virus types 6 and 11

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人乳头瘤病毒6型和11型通用型含内源性内参的重组酶介导的等温扩增技术检测方法的建立