Quantitative assay for group M (subtype A-H) and group OHIV-1 RNA detection in plasma

被引:7
作者
Ernest, I
Alexandre, I
Zammatteo, N
Herman, M
Houbion, A
De Lenner, F
Fransen, K
van der Groen, G
Remacle, J
机构
[1] Univ Namur, Dept Cellular Biochem & Biol, Namur, Belgium
[2] Lambdatech SA, Namur, Belgium
[3] Inst Trop Med, Dept Microbiol, B-2000 Antwerp, Belgium
关键词
HIV-1; competititive quantitative RT-PCR; internal standard;
D O I
10.1016/S0166-0934(00)00167-1
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A quantitative HIV-1 test is described based on a competitive RT-PCR assay combined with a sandwich hybridization as a detection system. The internal RNA standard (IS) was designed specifically to be competitive during the amplification and during the hybridization step. Sample viral load determination was carried out with one RT-PCR in the presence of 10(3) IS copies. The HIV-1 copy number was calculated by reference to an external standard curve performed on known and increasing amounts of the reference HIV-1 (Ref HIV-1) RNA co-amplified with a constant amount of the IS RNA. The assay had a linear range from 10(1) to 10(6) HIV-1 copies. HIV-I strains belonging to the different subtypes from group M, but also group O. were all detected. Absolute quantification of purified HIV-I RNA copies gave identical results as the AMPLICOR HIV-1 Monitor assay. The quantification of patient's samples was evaluated according to different criteria such as dynamic range, sensitivity. efficacy of material recovery, reproducibility and convenience of sample handling. The microplate format of the assay combined with the colorimetric detection provides a convenient tool and fulfills the requirement for routine molecular diagnostic laboratories. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:1 / 14
页数:14
相关论文
共 40 条
[1]   Subtype-specific problems with quantification of plasma HIV-1 RNA [J].
Alaeus, A ;
Lidman, K ;
Sonnerborg, A ;
Albert, J .
AIDS, 1997, 11 (07) :859-865
[2]   Quantitative determination of CMV DNA using a combination of competitive PCR amplification and sandwich hybridization [J].
Alexandre, I ;
Zammatteo, N ;
Ernest, I ;
Ladriere, JM ;
Le, L ;
Hamels, S ;
Chandelier, N ;
Vipond, B ;
Remacle, J .
BIOTECHNIQUES, 1998, 25 (04) :676-683
[3]   VIROLOGICAL AND IMMUNOLOGICAL CHARACTERIZATION OF LONG-TERM SURVIVORS OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 INFECTION [J].
CAO, YZ ;
QIN, LM ;
ZHANG, LQ ;
SAFRIT, J ;
HO, DD .
NEW ENGLAND JOURNAL OF MEDICINE, 1995, 332 (04) :201-208
[4]  
Coste J, 1997, J ACQ IMMUN DEF SYND, V15, P174
[5]   GENETIC-RELATIONSHIPS DETERMINED BY A DNA HETERODUPLEX MOBILITY ASSAY - ANALYSIS OF HIV-1 ENV GENES [J].
DELWART, EL ;
SHPAER, EG ;
LOUWAGIE, J ;
MCCUTCHAN, FE ;
GREZ, M ;
RUBSAMENWAIGMANN, H ;
MULLINS, JI .
SCIENCE, 1993, 262 (5137) :1257-1261
[6]   APPLICATION OF BRANCHED DNA SIGNAL AMPLIFICATION TO MONITOR HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 BURDEN IN HUMAN PLASMA [J].
DEWAR, RL ;
HIGHBARGER, HC ;
SARMIENTO, MD ;
TODD, JA ;
VASUDEVACHARI, MB ;
DAVEY, RT ;
KOVACS, JA ;
SALZMAN, NP ;
LANE, HC ;
URDEA, MS .
JOURNAL OF INFECTIOUS DISEASES, 1994, 170 (05) :1172-1179
[7]   DESIGN AND EVALUATION OF NEW, HIGHLY SENSITIVE AND SPECIFIC PRIMERS FOR POLYMERASE CHAIN-REACTION DETECTION OF HIV-1-INFECTED PRIMARY LYMPHOCYTES [J].
FRANSEN, K ;
ZHONG, P ;
DEBEENHOUWER, H ;
CARPELS, G ;
PEETERS, M ;
LOUWAGIE, J ;
JANSSENS, W ;
PIOT, P ;
VANDERGROEN, G .
MOLECULAR AND CELLULAR PROBES, 1994, 8 (04) :317-322
[8]  
Fransen K, 1998, AIDS, V12, P230
[9]   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
[10]   Reactivity and amplification efficiency of the NASBA HIV-1 RNA amplification system with regard to different HIV-1 subtypes [J].
Gobbers, E ;
Fransen, K ;
Oosterlaken, T ;
Janssens, WB ;
Heyndrickx, L ;
Ivens, T ;
Vereecken, K ;
Schoones, R ;
vandeWiel, P ;
vanderGreen, G .
JOURNAL OF VIROLOGICAL METHODS, 1997, 66 (02) :293-301