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用于鉴定肝炎病毒的序列特异性单引物扩增及PCR产物检测

Sequence-specific, single-primer amplification and detection of PCR products for identification of hepatitis viruses.

作者信息

Nainan O V, Cromeans T L, Margolis H S

机构信息

World Health Organization Collaborating Center for Research and Reference in Viral Hepatitis, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.

出版信息

J Virol Methods. 1996 Sep;61(1-2):127-34. doi: 10.1016/0166-0934(96)02077-0.

Abstract

A simple system to detect polymerase chain reaction (PCR) amplification products was developed. This detection method has the sensitivity and the specificity of nested primer PCR amplification or Southern blot hybridization of PCR product. Digoxigenin-labeled PCR products were hybridized with a biotinylated probe in liquid phase and captured on to microtiter wells coated with antidigoxigenin followed by detection with streptavidin-peroxidase. The sensitivity of this assay for the detection of hepatitis A virus, hepatitis B virus, and hepatitis C virus is equal to that of existing nucleic acid detection systems.

摘要

开发了一种用于检测聚合酶链反应(PCR)扩增产物的简单系统。这种检测方法具有巢式引物PCR扩增或PCR产物Southern印迹杂交的灵敏度和特异性。地高辛标记的PCR产物在液相中与生物素化探针杂交,然后捕获到包被抗地高辛的微量滴定孔上,接着用链霉抗生物素蛋白-过氧化物酶进行检测。该检测方法对甲型肝炎病毒、乙型肝炎病毒和丙型肝炎病毒的检测灵敏度与现有核酸检测系统相当。

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