Analysis of human immunodeficiency virus type 1 integration by using a specific, sensitive and quantitative assay based on real-time polymerase chain reaction

被引：48

作者：

Yamamoto, N

Tanaka, C

Wu, YF

Chang, MO

Inagaki, Y

Saito, Y

Naito, T

Ogasawara, H

Sekigawa, I

Hayashida, Y

机构：

[1] Dept Mol Virol, Bunkyo Ku, Tokyo 1138519, Japan

[2] Juntendo Univ, Sch Med, Dept Gen Med, Bunkyo Ku, Tokyo 1138421, Japan

[3] Juntendo Univ, Sch Med, Dept Rheumatol & Internal Med, Bunkyo Ku, Tokyo 1138421, Japan

[4] Juntendo Univ, Grad Sch Med, Inst Environm & Gender Specif Med, Chiba 2790021, Japan

来源：

VIRUS GENES | 2006年 / 32卷 / 01期

关键词：

HIV-1; integration; Alu-PCR; real-time PCR; nested PCR;

D O I：

10.1007/s11262-005-5851-2

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A novel real-time nested-PCR assay was developed to quantify integrated human immunodeficiency virus type-1 (HIV-1) DNA with high specificity and sensitivity. This assay reproducibly allowed the detection of three copies of integrated HIV DNA in a background of 100,000 cell equivalents of human chromosomal DNA. The non-specific amplification of unintegrated HIV-1 DNA was significantly inhibited in this assay and the specificity of this assay was much higher than the previously reported method. This assay showed that kinetics in viral DNA sysnthesis was cell-type dependent and that the kinetics of HIV-1 DNA integration was very rapid in Jurkat T cell line. This method may provide new insights into the integration processes and be useful in evaluating future integrase inhibitors.

引用

收藏

页码：105 / 113

页数：9

相关论文

共 20 条

[1] KINETIC-ANALYSIS OF HIV-1 EARLY REPLICATIVE STEPS IN A COCULTURE SYSTEM [J].

BARBOSA, P ;

CHARNEAU, P ;

DUMEY, N ;

CLAVEL, F .

AIDS RESEARCH AND HUMAN RETROVIRUSES, 1994, 10 (01) :53-59

[2] SOURCES AND EVOLUTION OF HUMAN ALU REPEATED SEQUENCES [J].

BRITTEN, RJ ;

BARON, WF ;

STOUT, DB ;

DAVIDSON, EH .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (13) :4770-4774

[3] Analysis of early human immunodeficiency virus type 1 DNA synthesis by use of a new sensitive assay for quantifying integrated provirus [J].

Brussel, A ;

Sonigo, P .

JOURNAL OF VIROLOGY, 2003, 77 (18) :10119-10124

[4] ASSOCIATION OF INTEGRASE, MATRIX, AND REVERSE-TRANSCRIPTASE ANTIGENS OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 WITH VIRAL NUCLEIC-ACIDS FOLLOWING ACUTE INFECTION [J].

BUKRINSKY, MI ;

SHAROVA, N ;

MCDONALD, TL ;

PUSHKARSKAYA, T ;

TARPLEY, WG ;

STEVENSON, M .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (13) :6125-6129

[5] ACTIVE NUCLEAR IMPORT OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PREINTEGRATION COMPLEXES [J].

BUKRINSKY, MI ;

SHAROVA, N ;

DEMPSEY, MP ;

STANWICK, TL ;

BUKRINSKAYA, AG ;

HAGGERTY, S ;

STEVENSON, M .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (14) :6580-6584

[6] Human immunodeficiency virus cDNA metabolism: Notable stability of two-long terminal repeat circles [J].

Butler, SL ;

Johnson, EP ;

Bushman, FD .

JOURNAL OF VIROLOGY, 2002, 76 (08) :3739-3747

[7] A quantitative assay for HIV DNA integration in vivo [J].

Butler, SL ;

Hansen, MST ;

Bushman, FD .

NATURE MEDICINE, 2001, 7 (05) :631-634

[8] Presence of an inducible HIV-1 latent reservoir during highly active antiretroviral therapy [J].

Chun, TW ;

Stuyver, L ;

Mizell, SB ;

Ehler, LA ;

Mican, JAM ;

Baseler, M ;

Lloyd, AL ;

Nowak, MA ;

Fauci, AS .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (24) :13193-13197

[9] INTEGRATION IS REQUIRED FOR PRODUCTIVE INFECTION OF MONOCYTE-DERIVED MACROPHAGES BY HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 [J].

ENGLUND, G ;

THEODORE, TS ;

FREED, EO ;

ENGELMAN, A ;

MARTIN, MA .

JOURNAL OF VIROLOGY, 1995, 69 (05) :3216-3219

[10] HIV-1 cDNA integration: Requirement of HMG I(Y) protein for function of preintegration complexes in vitro [J].

Farnet, CM ;

Bushman, FD .

CELL, 1997, 88 (04) :483-492