基因扩增产物的固相杂交-酶联显色方法的建立  

Quantification of Polymerase Chain Reaction Products Using Solid Phase Hybridization Enzyme Colorimetric Detection

作　　者：王凤水[1] 王宇[1] 陈红松[1] 丛旭[1] 

机构地区：[1]北京医科大学人民医院肝病研究所

出　　处：《中国生物化学与分子生物学报》1998年第2期192-198,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金

摘　　要：建立基于基因扩增技术的简便、快速的病毒核酸定量检测方法．将标记有生物素的寡核苷酸引物所扩增的病毒基因产物，与通过共价键结合在微孔反应板上的特异性探针进行快速杂交，然后通过辣根过氧化物酶标记的抗生物素进行酶联显色，读取光密度值．应用本方法对血清中乙型、丙型肝炎病毒核酸定量检测，灵敏度分别可达１－５拷贝／反应．此方法简便、快速、特异性好、敏感性高、半定量指标客观，可广泛应用于肝炎病毒感染的临床诊断和疗效评价．A simple,rapid polymerase chain reaction(PCR) based on DNA quantification technique was established.For detecting specific viral nucleic acids,the primer labeled with biotin was used to amplify viral gene fragment.Then the 1st PCR product was complemently hybridized with the specific probe covalently coupled onto microplate wells.Finally,Streptavidin POD was used in colorimetric detection.With this method,the sensitivity of the detection system could be 1—5 copies of hepatitis B genome and hepatitis C genome respectively.Simple,rapid with high specificity,sensitivity and semiquantification,the method described can be of general application for detection of any foreign pathogen in blood,other body fluids and tissue samples,used both in clinical diagnosis and estimate of the efficacy of antiviral therapy.

关 键 词：核酸定量 固相杂交 酶联显色 PCR 肝炎病毒 

分 类 号：Q52[生物学—生物化学] R512.6[医药卫生—内科学]