微流控芯片检测病原体基因pcr扩增产物

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1、_______________________________________________________________________________www.paper.edu.cn微流控芯片检测病原体基因PCR扩增产物刘兆奕秦建华王鸾升[摘要]目的以微流控芯片为平台以激光诱导荧光为检测手段对临床常见的病原体结核tuberculosisTB和丙型肝炎hepatitisCvirusHCV基因扩增产物进行快速检测目的在于为临床病原体检测建立一种高效快速灵敏的分析方法方法利用微流控芯片分析系统对临床疑诊的15例TB患

2、者待检痰液标本与32例HCV患者的待检血清标本中的病原体结核杆菌和丙型肝炎病毒基因扩增产物进行检测并与临床上已有的检测方法杂交梳和实时荧光PCR的方法real-timePCR进行比较结果TB基因特异性扩增片段240bp与HCV基因特异性扩增片段93bp分别在270S210s内得到分离检测具有100%的灵敏度2和特异性且检测限接近是杂交梳的12800倍实时荧光PCR法的检测限是510拷贝2微升而芯片电泳检测限是0.0312510拷贝微升结论微流控芯片用于临床常见的病原体基因检测具有结果直观分析时间短操作简单高效灵敏方法经

3、济及易于实现高通量等特点可为临床病原体的检测提供一方便灵活的操作平台[关键词]微流控芯片病原体丙型肝炎结核ThedetectionofthePCRproductsofclinicalpathogenbymicrochipelectrophoresis.LIUZhao-yi,QINJian-hua,WANGLuan-sheng.DepartmentofPediatrics,TheFirstHospitalDaLianMedicalUniversity,DaLian116027,China.[Abstract]Object

4、iveSensitivedetectionofthePCRproductsoftuberculosisandhepatitisCvirusgenefromclinicalspecimensbymicrochipelectrophoresis.MethodPCRproductsfrom15selectedTBand32HCVspecimenswhichsubmittesforTBandHCVdetectionwereanalyzedbymicrochipelectrophoresisandtheestablishedcl

5、inicalmethod.ResultsAnalysistimefordetectionofTBDNAbyPCR-Hyricombwas2-3hour.While,Microchipelectrophoresisprovidedidenticalresultsin270secondpersample.Microchipelectrophoresisresolvedthe93bpofHCVin210second,achieving100%sensitivityandspecificitycomparedwiththees

6、tablishedmethod.Thedetectionlimitofmicrochipwasapproximately12800timeshigherthanPCR-Hyricomb.The2detectionlimitofmicrochipwas0.0312510copiesperllcomparedwithThedetectionlimit2of510copiesperllonrealtimePCR.ConclusionMicrochipelectrophoresiscanrapidlyandaccurately

7、separateTBandHCVPCRproducts,givingresultsidenticaltothoseobstainedbytheestablishedclinicalmethods.Butithasthefollowingadvantagessuchashighresolutionrate,highsensitivity,lowreagentconsumptionandeasytohighthroughput,etc.Accordingtotheabovefeature,microchipelectrop

8、horesisissuitableforclinicalanalysis.[Keywords]MicrochipelectrophoresisPathogenHepatitisCvirusTuberculosis结核tuberculosisTB是长期以来危害人类生命健康的严重疾病在发展中国家结核病迄今仍是常见病结核病的诊断主要根据