使用聚合酶链反应和比色酶检测系统检测和定量血清中的丙型肝炎病毒RNA。

Detection and quantitation of hepatitis C virus RNA in serum using the polymerase chain reaction and a colorimetric enzymatic detection system.

作者信息

Shindo M, Di Bisceglie A M, Silver J, Limjoco T, Hoofnagle J H, Feinstone S M

机构信息

Hepatitis Studies Section, NIH, NIDDK, Bethesda, MD.

出版信息

J Virol Methods. 1994 Jun;48(1):65-72. doi: 10.1016/0166-0934(94)90089-2.

DOI:10.1016/0166-0934(94)90089-2

PMID:7962261

Abstract

A sensitive, non-isotopic method for detecting and quantifying hepatitis C virus (HCV) RNA in serum using the reverse transcriptase-polymerase chain reaction (RT-PCR) and a hybridization specific, colorimetric biotin-avidin peroxidase detection system has been developed. The sensitivity of the PCR-colorimetric system was determined using RNA synthesized from cloned HCV cDNA. The assay could detect as few as 10 molecules of HCV RNA, comparable to the sensitivity achieved with double PCR using nested primers. Thus, this colorimetric assay can detect low levels of HCV RNA in serum and appears to be quantitative, suggesting that this technique may be applied to rapid screening of large numbers of samples and to monitor the effect of antiviral therapy.

摘要

已开发出一种灵敏的非同位素方法，该方法利用逆转录聚合酶链反应（RT-PCR）以及杂交特异性比色生物素-抗生物素蛋白过氧化物酶检测系统，对血清中的丙型肝炎病毒（HCV）RNA进行检测和定量。使用从克隆的HCV cDNA合成的RNA来确定PCR比色系统的灵敏度。该检测方法能够检测低至10个HCV RNA分子，与使用巢式引物的双重PCR所达到的灵敏度相当。因此，这种比色检测法能够检测血清中低水平的HCV RNA，并且似乎具有定量功能，这表明该技术可应用于大量样本的快速筛查以及监测抗病毒治疗的效果。