一种快速简单的可携带式高危型人乳头瘤病毒现场分子诊断装置与方法  

A rapid, simple and field-deployable point-of-care molecular detection method for high-risk human papillomavirus

作　　者：梅俊杨 王丹丹 张忆恒 崔金辉 甘明哲[2] 刘培峰[1,3] MEI Junyang;WANG Dandan;ZHANG Yiheng;CUI Jinhui;GAN Mingzhe;LIU Peifeng(State Key Laboratory of Oncogenes and Related Genes,Shanghai Cancer Institute,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200032,China;Chinese Academy of Sciences Key Laboratory of Nano-Bio Interface,Suzhou Institute of Nano-Tech and Nano-Bionics,Chinese Academy of Sciences,Suzhou 215123,Jiangsu Province,China;Central Laboratory,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China)

机构地区：[1]上海交通大学医学院附属仁济医院上海市肿瘤研究所癌基因与相关基因国家重点实验室,上海200032 [2]中国科学院苏州纳米技术与纳米仿生研究所中国科学院纳米-生物界面重点实验室,江苏苏州215123 [3]上海交通大学医学院附属仁济医院中心实验室,上海200127

出　　处：《肿瘤》2022年第7期508-517,共10页Tumor

基　　金：国家自然科学基金(81771968)

摘　　要：目的:构建一种快速简单的高危型人乳头瘤病毒(high-risk human papillomavirus,hr-HPV)现场诊断方法,以期加强对hr-HPV感染的监测及其相关癌症的预防和早期筛查。方法:以重组酶聚合酶扩增(recombinase polymerase amplification,RPA)反应为基础,建立一种快速且准确的可视化hr-HPV检测方法。采用纸基富集法提取样本中的HPV-DNA,构建恒温孵育与结果读取一体化的充电式便携阅读器,并且使用hr-HPV模拟病毒样本评估本方法的性能。结果:该方法实现了30 min内的样本进-结果出的免仪器检测,并且能够输出可视化结果。对hr-HPV模拟病毒样本的检测灵敏度可达10~4 copies/mL,并且稳定性和特异度均表现良好。结论:该方法使用简单且性能优越,可以根据需要广泛部署于靠近患者的护理点以及社区医院、诊所和药房等基层卫生服务设施,有助于加强对hr-HPV感染的监测及其相关癌症的预防和早期筛查。Objective:To develop a rapid,simple and point-of-care testing(POCT)method for high-risk human papillomavirus(hr-HPV)to promote the monitoring of hr-HPV infection and the prevention and early screening of related cancers.Methods:A rapid and accurate visualized detection method for hr-HPV based on recombinase polymerase amplification(RPA)was established.A paper-based enrichment method was used to extract HPV-DNA,and a rechargeable portable reader was constructed for integrated isothermal incubation and result readout.The performance of this POCT method was evaluated by using hr-HPV contrived samples.Results:Instrument-free and sample-to-answer detection within 30 minutes was achieved,and outputs were visualized.This POCT method has demonstrated a minimum detection threshold of 10~4 copies/mL for hr-HPV contrived samples with excellent stability and specificity.Conclusion:This POCT method is easy-to-use and has superior performance.It can be widely deployed in POCT environments as well as in primary health service facilities,such as community hospitals,clinics and pharmacies,to promote the monitoring of hr-HPV infection and the prevention and early screening of the related cancers.

关 键 词：致癌性感染 高危型人乳头瘤病毒 宫颈癌 即时检验 纸基核酸富集 可现场部署的 可视化检测 

分 类 号：R737.33[医药卫生—肿瘤] R446[医药卫生—临床医学]