A polymerase chain reaction method for the amplification of full-length envelope genes of HIV-1 from DNA samples containing single molecules of HIV-1 provirus

被引：9

作者：

McClure, P ^{[1
]}

Curran, R ^{[1
]}

Boneham, S ^{[1
]}

Ball, JK ^{[1
]}

机构：

[1] Univ Nottingham, Queens Med Ctr, Div Microbiol & Infect Dis, Nottingham NG7 2UH, England

来源：

JOURNAL OF VIROLOGICAL METHODS | 2000年 / 88卷 / 01期

基金：

英国医学研究理事会;

关键词：

limiting dilution PCR; HIV-1; envelope; gp160;

D O I：

10.1016/S0166-0934(00)00179-8

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Polymerase chain reaction (PCR) amplification of full-length envelope genes from the human immunodeficiency virus type 1 (HIV-1) directly from uncultured clinical samples is difficult. This paper describes a comparative assessment of the performance of three thermostable polymerases in an HIV-1 full-length envelope gene PCR. The PCR method utilising Expand HiFi polymerase was successful when using DNA samples extracted from a variety of sources including blood, semen and various tissues. This method generated high and specific yields of product from samples containing as little as one copy of HIV-1 proviral DNA. The resulting PCR products were suitable for a variety of downstream analytical methods including DNA sequence analysis. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

收藏

页码：73 / 80

页数：8

相关论文

共 21 条

[1]

Abu Al-Soud W, 1998, APPL ENVIRON MICROB, V64, P3748

[2] Fidelity and mutational spectrum of Pfu DNA polymerase on a human mitochondrial DNA sequence [J].

Andre, P ;

Kim, A ;

Khrapko, K ;

Thilly, WG .

GENOME RESEARCH, 1997, 7 (08) :843-852

[3] Production of single-stranded DNA using a uracil-N-glycosylase-mediated asymmetric polymerase chain reaction method [J].

Ball, JK ;

Curran, R .

ANALYTICAL BIOCHEMISTRY, 1997, 253 (02) :264-267

[4] GENOMIC VARIATION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 (HIV-1) - MOLECULAR ANALYSES OF HIV-1 IN SEQUENTIAL BLOOD-SAMPLES AND VARIOUS ORGANS OBTAINED AT AUTOPSY [J].

BALL, JK ;

HOLMES, EC ;

WHITWELL, H ;

DESSELBERGER, U .

JOURNAL OF GENERAL VIROLOGY, 1994, 75 :867-879

[5] Contribution of Taq polymerase-induced errors to the estimation of RNA virus diversity [J].

Bracho, MA ;

Moya, A ;

Barrio, E .

JOURNAL OF GENERAL VIROLOGY, 1998, 79 :2921-2928

[6] PCR fidelity of Pfu DNA polymerase and other thermostable DNA polymerases [J].

Cline, J ;

Braman, JC ;

Hogrefe, HH .

NUCLEIC ACIDS RESEARCH, 1996, 24 (18) :3546-3551

[7] An efficient method for the rescue and analysis of functional HIV-1 env genes: Evidence for recombination in the vicinity of the tat/rev splice site [J].

Douglas, NW ;

Knight, AI ;

Hayhurst, A ;

Barrett, WY ;

Kevany, MJ ;

Daniels, RS .

AIDS, 1996, 10 (01) :39-46

[8] A comprehensive panel of near-full-length clones and reference sequences for non-subtype B isolates of human immunodeficiency virus type 1 [J].

Gao, F ;

Robertson, DL ;

Carruthers, CD ;

Morrison, SG ;

Jian, BX ;

Chen, YL ;

Barré-Sinoussi, F ;

Girard, M ;

Srinivasan, A ;

Abimiku, AG ;

Shaw, GM ;

Sharp, PM ;

Hahn, BH .

JOURNAL OF VIROLOGY, 1998, 72 (07) :5680-5698

[9] Molecular cloning and analysis of functional envelope genes from human immunodeficiency virus type 1 sequence subtypes A through G [J].

Gao, F ;

Morrison, SG ;

Robertson, DL ;

Thornton, CL ;

Craig, S ;

Karlsson, G ;

Sodroski, J ;

Morgado, M ;

GalvaoCastro, B ;

vonBriesen, H ;

Beddows, S ;

Weber, J ;

Sharp, PM ;

Shaw, GM ;

Hahn, BH ;

Osmanov, S ;

Heyward, WL ;

Esparza, J ;

vandePerre, P ;

Karita, E ;

Sempala, S ;

Tugume, B ;

Biryahwaho, B ;

Wasi, C ;

RubsamenWaigmann, H ;

Holmes, H ;

Newberry, A ;

Ranjbar, S ;

Tomlinson, P ;

Bradac, J ;

Mullins, JI ;

Delwart, EL ;

CheingsongPopov, R ;

Kaleebu, P ;

Myers, G ;

Korber, BTM ;

Chiphangwi, J ;

Taha, T ;

Desormeaux, J ;

Eiumtrakul, S ;

Natpratan, C ;

Khamboonruang, C ;

Miotti, P ;

Halsey, NA ;

Vlahov, D ;

Nelson, KE ;

Phair, J ;

Cao, Y ;

Moore, JP ;

Ho, DD .

JOURNAL OF VIROLOGY, 1996, 70 (03) :1651-1667

[10]

Günther S, 1998, J CLIN MICROBIOL, V36, P531