交叉置换扩增联合纳米生物传感器检测乙肝病毒方法的建立与应用  

Development and application of multiple cross displacement amplification coupled with label-based gold nanoparticles lateral flow biosensor for detection of hepatitis B virus

作　　者：贾楠 肖飞[1] 周娟 伏瑾[1] 黄小兰[1] 陈敏 黄辉[2] 王毅 Jia Nan;Xiao Fei;Zhou Juan;Fu Jin;Huang Xiaolan;Chen Min;Huang Hui;Wang Yi(Experimental Research Center,Capital Institute of Pediatrics,Beijing 100020,China;Department of Infectious Diseases,Affiliated Children’s Hospital,Capital Institute of Pediatrics,Beijing 100020,China)

机构地区：[1]首都儿科研究所实验中心,北京100020 [2]首都儿科研究所附属儿童医院感染科,北京100020

出　　处：《遵义医科大学学报》2023年第7期704-711,共8页Journal of Zunyi Medical University

基　　金：国家重点研发计划项目(NO:2021YFC2301102)。

摘　　要：目的建立一种敏感、特异的乙肝病毒(HBV)快速检测方法,并评价其应用。方法基于多交叉置换扩增(MCDA)技术和纳米生物传感(LFB)检测技术,以HBV特异性S基因为检测靶标,建立HBV-MCDA-LFB方法,优化反应温度和时间等实验条件,并应用临床样本进行评价,同时与实时PCR(RT-PCR)进行比较。结果HBV-MCDA-LFB能在30 min内完成HBV检测,最佳反应温度为60℃,其特异性良好,未与其他病毒、细菌发生交叉反应。此外,该方法的检出限低至1×101拷贝/反应,且能准确检测临床样本的HBV,检测灵敏度优于RT-PCR方法。结论本研究建立了一种特异、敏感、便捷的乙肝病毒快速检测方法,即HBV-MCDA-LFB,其诊断效率优于临床采用的RT-PCR方法,可作为临床快速诊断HBV感染的替代方法。Objective To establish a sensitive and specific method for rapid detection of hepatitis B virus(HBV)and evaluate its clinical application.Methods By coupling multiple cross displacement amplification(MCDA)with label-based gold nanoparticles lateral flow biosensor(LFB),a novel method targeting on the specific S gene of HBV was developed and namedas HBV-MCDA-LFB.The optimal reaction time and temperature of HBV-MCDA-LFB were recognized,and its performance in clinical samples was demonstrated via comparing to that of real-time PCR(RT-RCR).Results The newly developed HBV-MCDA-LFB method showed high specificity and could rapidly accomplish HBV detection in about 30 minutes.This assay was optimally performed at 60℃with a limit of detection(LoD)of 1×101 copies,and exhibited superior sensitivity comparedto RT-RCR in clinical samples.Conclusion With the merits of high sensitivity and specificity,and easy-to-use,HBV-MCDA-LFB surpassed the clinically widely used RT-RCR method and was expected to be a promising substitute for rapid diagnosis of HBV infection in clinical settings.

关 键 词：乙肝病毒 多交叉置换扩增 纳米生物传感 HBV-MCDA-LFB RT-PCR 

分 类 号：R512.6[医药卫生—内科学]