A SINGLE TUBE NESTED PCR FOR THE DETECTION OF HEPATITIS-C VIRUS-RNA

被引:17
作者
TILSTON, P
CORBITT, G
机构
[1] Clinical Virology, Manchester Central Laboratory Services, Manchester Royal Infirmary, Manchester, M13 9WL, Clinical Sciences Building
关键词
SINGLE TUBE NESTED PCR; COMBINED RT/PCR; HEPATITIS C VIRUS (HCV);
D O I
10.1016/0166-0934(95)00006-G
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An assay was developed for the detection of hepatitis C virus RNA in serum which combined cDNA synthesis and a hot start nested polymerase chain reaction (PCR) in a single tube. This was made possible by separation of the reagents necessary for cDNA synthesis and PCR during cDNA synthesis with a high melting temperature wax interface and by the use of 'drop in-drop out' nested primers which enabled each primer set to be selectively extended within the same reaction tube. The increase in sensitivity following amplification with the internal primer pair was comparable to that achieved when nested reactions are carried out separately.
引用
收藏
页码:121 / 129
页数:9
相关论文
共 23 条
[1]  
AATSINKI JT, 1994, BIOTECHNIQUES, V16, P48
[2]   SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159
[3]   GENETIC ORGANIZATION AND DIVERSITY OF THE HEPATITIS-C VIRUS [J].
CHOO, QL ;
RICHMAN, KH ;
HAN, JH ;
BERGER, K ;
LEE, C ;
DONG, C ;
GALLEGOS, C ;
COIT, D ;
MEDINASELBY, A ;
BARR, PJ ;
WEINER, AJ ;
BRADLEY, DW ;
KUO, G ;
HOUGHTON, M .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (06) :2451-2455
[4]   RECENT ADVANCES IN THE POLYMERASE CHAIN-REACTION [J].
ERLICH, HA ;
GELFAND, D ;
SNINSKY, JJ .
SCIENCE, 1991, 252 (5013) :1643-1651
[5]   REINFECTION OF LIVER GRAFT BY HEPATITIS-C VIRUS AFTER LIVER-TRANSPLANTATION [J].
FERAY, C ;
SAMUEL, D ;
THIERS, V ;
GIGOU, M ;
PICHON, F ;
BISMUTH, A ;
REYNES, M ;
MAISONNEUVE, P ;
BISMUTH, H ;
BRECHOT, C .
JOURNAL OF CLINICAL INVESTIGATION, 1992, 89 (04) :1361-1365
[6]   IMPROVEMENT OF HCV GENOME DETECTION WITH SHORT PCR PRODUCTS [J].
GARSON, JA ;
RING, CJA ;
TUKE, PW .
LANCET, 1991, 338 (8780) :1466-1467
[7]   DETECTION OF HEPATITIS-C VIRAL SEQUENCES IN BLOOD DONATIONS BY NESTED POLYMERASE CHAIN-REACTION AND PREDICTION OF INFECTIVITY [J].
GARSON, JA ;
TEDDER, RS ;
BRIGGS, M ;
TUKE, P ;
GLAZEBROOK, JA ;
TRUTE, A ;
PARKER, D ;
BARBARA, JAJ ;
CONTRERAS, M ;
ALOYSIUS, S .
LANCET, 1990, 335 (8703) :1419-1422
[8]  
Gelfand D.H., 1990, PC PRO, P129
[9]   DETECTION OF HEPATITIS-C VIRUS-RNA - COMPARISON OF ONE-STAGE POLYMERASE CHAIN-REACTION (PCR) WITH NESTED-SET PCR [J].
GRETCH, DR ;
WILSON, JJ ;
CARITHERS, RL ;
DELAROSA, C ;
HAN, JH ;
COREY, L .
JOURNAL OF CLINICAL MICROBIOLOGY, 1993, 31 (02) :289-291
[10]  
HAYASHI Y, 1994, RES VIROLOGY, V145, P1213