COLORIMETRIC DETECTION OF COMPETITIVE PCR PRODUCTS FOR QUANTIFCATION OF HEPATITIS-C VIREMIA

被引：24

作者：

YUN, ZB

LUNDEBERG, J

JOHANSSON, B

HEDRUM, A

WEILAND, O

UHLEN, M

SONNERBORG, A

机构：

[1] STOCKHOLM CTY COUNCIL,CENT MICROBIOL LAB,DEPT VIROL,S-10726 STOCKHOLM,SWEDEN

[2] ROYAL INST TECHNOL,DEPT BIOCHEM & BIOTECHNOL,S-10044 STOCKHOLM,SWEDEN

[3] KAROLINSKA INST,HUDDINGE HOSP,DEPT INFECT DIS,S-10401 STOCKHOLM,SWEDEN

来源：

JOURNAL OF VIROLOGICAL METHODS | 1994年 / 47卷 / 1-2期

关键词：

COMPETITIVE PCR; COLORIMETRIC DETECTION; HCV GENOME; LAC OPERATOR; INTERFERON; QUANTIFICATION;

D O I：

10.1016/0166-0934(94)90061-2

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method based on competitive polymerase chain reaction (PC R) and colorimetric detection of the amplified products was developed to quantify hepatitis C virus (HCV) genomes. Serum samples were obtained from patients who were treated with interferon alpha (IFN-alpha). After reverse transcription of the HCV RNA, the cDNA was coamplified with a serially diluted cloned HCV competitor DNA using nested PCR. The competitor DNA consisted of the amplified region of the wild type HCV cDNA with an internal region substituted with the lac operator (lacO) sequence. The PCR products were quantitated specifically by a colorimetric solid-phase assay. The results suggest that the method is well suited for analysing the kinetics of the anti-HCV effects during IFN-alpha treatment. The quantification assay is simple, reliable and suitable for quantitating HCV genomes in a large number of clinical samples.

引用

页码：1 / 13

页数：13

共 17 条

[1] EFFECT OF ALPHA-INTERFERON THERAPY ON HEPATITIS-C VIREMIA IN COMMUNITY-ACQUIRED CHRONIC NON-A-HEPATITIS, NON-B-HEPATITIS - A QUANTITATIVE POLYMERASE CHAIN-REACTION STUDY [J].