肽核酸生物传感器直接检测临床标本中提取的病毒基因组DNA  

Direct Detection of Genomic DNA of Virus from Clinical Samples with PNA Biosensor

作　　者：陈鸣[1] 府伟灵[1] 蔡国儒[2] 刘明华[1] 张波[1] 蒋天伦[1] 陈庆海[1] 秦廷辉[2] 

机构地区：[1]第三军医大学附属西南医院,重庆400038 [2]中国电子科技集团公司第26研究所,重庆400060

出　　处：《中华医院感染学杂志》2003年第7期616-619,共4页Chinese Journal of Nosocomiology

基　　金：国家 8 6 3计划 (2 0 0 2 AARZ2 0 2 3);国家"九五"科技攻关项目 (96 - A2 3- 0 4 - 0 4 );国家自然科学基金 (30 2 70 388);军队"九五"课题 (98MO94 );重庆市应用基础研究项目 (2 0 0 2 - 10 - 78);国家高技术计划生物技术领域青年基金;重庆市科委风

摘　　要：目的　以肽核酸生物传感器检测系统直接检测从临床标本中提取的乙型肝炎病毒基因组 DNA。方法　抽取 6 3例临床免疫学检测证实为乙型肝炎病毒 (HBV)感染者标本血清的 DNA,以及 15例免疫学检测全阴的标本血清中的 DNA,分别用肽核酸生物传感器法以及定量 PCR法检测 ,比较两种方法的优劣并确定传感器法的检测限。结果　传感器法检测 6 0例标本阳性 ,阳性率为 95 .2 4 % ,而定量 PCR法检测 6 3例均为阳性 ;15例阴性血清两种方法检测均为阴性 ;传感器法的检测限为 8.6 pg/ L ;两种方法检测 HBV差异无显著性 ,但传感器检测法所用时间更短 ,且无需进行探针的标记。结论　利用肽核酸生物传感器成功地绕过了 PCR扩增而直接检测出了临床标本中的 HBV基因组 DNA。OBJECTIVE To directly detect HBV genomic DNA extracted from clinical samples with peptide nucleic acid (PNA) biosensor. METHODS Extracted genomic DNA from 63 clinical positive samples caught from hepatitis patients and 15 negative sera samples were detected with PNA biosensor and quantitative PCR, respectively. The advantages and disadvantages of both methods were summarized and the detection limit of sensor method was finally determined. RESULTS Sixty samples were positive among 63 clinical cases with hepatitis detected by PNA biosensor, and positive rate hence was 95.24%. While 63 clinical cases were all positive using quantitative PCR. And 15 negative clinical cases were all undetected by both two methods. Detection limit of biosensor was 8.6 pg/L. No significant difference existed between two methods according to statistic analysis, but less time was used and no probe should be marked at biosensor method. CONCLUSIONS HBV DNA extracted from clinical samples were directly detected using PNA biosensor and PCR amplification was successfully bypassed.

关 键 词：肽核酸 生物传感器 乙型肝炎病毒 

分 类 号：R446.8[医药卫生—诊断学]