实时定量聚合酶链反应与其他三种丙型肝炎病毒定量检测方法的比较。

Comparison of real-time quantitative polymerase chain reaction with three other assays for quantitation of hepatitis C virus.

作者信息

Enomoto M, Nishiguchi S, Shiomi S, Tanaka M, Fukuda K, Ueda T, Tamori A, Habu D, Takeda T, Yano Y, Otani S

机构信息

Third Department of Internal Medicine, Osaka City University Medical School, Osaka, Japan.

出版信息

J Gastroenterol Hepatol. 2001 Aug;16(8):904-9. doi: 10.1046/j.1440-1746.2001.02542.x.

DOI:10.1046/j.1440-1746.2001.02542.x

PMID:11555105

Abstract

BACKGROUND AND AIMS

Evaluation of serum levels of hepatitis C virus (HCV) is important for predicting the response to interferon treatment and monitoring its therapeutic efficacy. The aim of this study was to evaluate real-time quantitative polymerase chain reaction (PCR) as a method for the measurement of HCV-RNA.

METHODS

The subjects were 50 patients with chronic hepatitis C: 36 with genotype 1b, eight with genotype 2a, and six with genotype 2b. Samples were tested for HCV-RNA by using real-time quantitative PCR with the ABI Prism 7700 sequence detection system, a branched DNA signal amplification assay, and an Amplicor monitor test; and for HCV core protein by using a fluorescent enzyme immunoassay.

RESULTS

The detection range of the real-time quantitative PCR was between 10(1)-10(8) copies/mL of HCV-RNA. Hepatitis C virus RNA was detectable in all 50 samples by the use of real-time quantitative PCR, but was undetectable in 14 samples by the use of a branched DNA assay and in two samples by using the Amplicor monitor test; HCV core protein was undetectable in three samples. A significant correlation was found between the results of real-time quantitative PCR and those of the three other assays: branched DNA assay (r = 0.837, P < 0.0001), Amplicor monitor test (r = 0.853, P < 0.0001), and HCV core protein concentrations (r = 0.549, P < 0.0001).

CONCLUSIONS

Our results showed that the real-time quantitative PCR was a highly sensitive assay for the measurement of HCV-RNA.

摘要

背景与目的

评估丙型肝炎病毒（HCV）血清水平对于预测干扰素治疗反应及监测其治疗效果至关重要。本研究旨在评估实时定量聚合酶链反应（PCR）作为测量HCV-RNA的一种方法。

方法

研究对象为50例慢性丙型肝炎患者，其中36例为1b基因型，8例为2a基因型，6例为2b基因型。采用ABI Prism 7700序列检测系统的实时定量PCR、分支DNA信号放大检测法及Amplicor监测试验检测样本中的HCV-RNA；采用荧光酶免疫分析法检测HCV核心蛋白。

结果

实时定量PCR的检测范围为10(1)-10(8)拷贝/mL的HCV-RNA。使用实时定量PCR时，所有50个样本中均可检测到丙型肝炎病毒RNA，但使用分支DNA检测法时，14个样本未检测到，使用Amplicor监测试验时，2个样本未检测到；3个样本中未检测到HCV核心蛋白。实时定量PCR结果与其他三种检测结果之间存在显著相关性：分支DNA检测法（r = 0.837，P < 0.0001）、Amplicor监测试验（r = 0.853，P < 0.0001）及HCV核心蛋白浓度（r = 0.549，P < 0.0001）。