SENSITIVE NONRADIOACTIVE DETECTION OF HIV-1 - USE OF NESTED PRIMERS FOR THE AMPLIFICATION OF HIV DNA

被引:5
作者
TEGLBJAERG, LLS [1 ]
NIELSEN, C [1 ]
HANSEN, JES [1 ]
机构
[1] STATE SERUM INST, DEPT VIROL, DK-2300 COPENHAGEN, DENMARK
关键词
POLYMERASE CHAIN REACTION; HUMAN IMMUNODEFICIENCY VIRUS; NESTED PRIMERS; BIOTIN LABELED PROBE;
D O I
10.1016/0890-8508(92)90014-O
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This report describes the use of the polymerase chain reaction (PCR) for the non-radioactive detection of HIV-1 proviral genomic sequences in HIV-1 infected cells. We have developed a sensitive assay, using three different sets of nested primers and our results show that this method is superior to standard PCR for the detection of HIV-1 DNA. The assay described features the use of a simple and inexpensive sample preparation technique and a non-radioactive hybridization procedure for confirmation of results. To test the suitability of the assay for clinical purposes, we tested cell samples from 76 anti-HlV-1 positive patients. All were positive for at least one primer set: 88% were positive for all three sets of primers; 9% were positive for two sets of primers and 3% were positive for only one set of primers. It provides a useful approach to the study of HIV-1 infection in patient samples where genomic copies often are present at such low numbers that they are otherwise undetectable. © 1992.
引用
收藏
页码:175 / 180
页数:6
相关论文
共 18 条
[1]   SIMPLE, SENSITIVE, AND SPECIFIC DETECTION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 IN CLINICAL SPECIMENS BY POLYMERASE CHAIN-REACTION WITH NESTED PRIMERS [J].
ALBERT, J ;
FENYO, EM .
JOURNAL OF CLINICAL MICROBIOLOGY, 1990, 28 (07) :1560-1564
[2]   ISOLATION OF LYMPHOCYTES, GRANULOCYTES AND MACROPHAGES [J].
BOYUM, A .
SCANDINAVIAN JOURNAL OF IMMUNOLOGY, 1976, :9-15
[4]   BIOLOGICAL AND BIOCHEMICAL-CHARACTERIZATION OF A CLONED LEU-3- CELL SURVIVING INFECTION WITH THE ACQUIRED-IMMUNE-DEFICIENCY-SYNDROME RETROVIRUS [J].
FOLKS, TM ;
POWELL, D ;
LIGHTFOOTE, M ;
KOENIG, S ;
FAUCI, AS ;
BENN, S ;
RABSON, A ;
DAUGHERTY, D ;
GENDELMAN, HE ;
HOGGAN, MD ;
VENKATESAN, S ;
MARTIN, MA .
JOURNAL OF EXPERIMENTAL MEDICINE, 1986, 164 (01) :280-290
[5]   DETECTION OF LYMPHOCYTES EXPRESSING HUMAN LYMPHOTROPIC-T VIRUS TYPE-III IN LYMPH-NODES AND PERIPHERAL-BLOOD FROM INFECTED INDIVIDUALS BY INSITU HYBRIDIZATION [J].
HARPER, ME ;
MARSELLE, LM ;
GALLO, RC ;
WONGSTAAL, F .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (03) :772-776
[6]   A REPLICATION-DEFICIENT HIV-1 DNA USED FOR QUANTITATION OF THE POLYMERASE CHAIN-REACTION (PCR) [J].
HART, C ;
CHANG, SY ;
KWOK, S ;
SNINSKY, J ;
OU, CY ;
SCHOCHETMAN, G .
NUCLEIC ACIDS RESEARCH, 1990, 18 (13) :4029-4030
[7]   RAPID AND SENSITIVE METHOD FOR THE DETECTION OF SERUM HEPATITIS-B VIRUS-DNA USING THE POLYMERASE CHAIN-REACTION TECHNIQUE [J].
KANEKO, S ;
FEINSTONE, SM ;
MILLER, RH .
JOURNAL OF CLINICAL MICROBIOLOGY, 1989, 27 (09) :1930-1933
[8]   AVOIDING FALSE POSITIVES WITH PCR [J].
KWOK, S ;
HIGUCHI, R .
NATURE, 1989, 339 (6221) :237-238
[9]   EFFECTS OF PRIMER TEMPLATE MISMATCHES ON THE POLYMERASE CHAIN-REACTION - HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 MODEL STUDIES [J].
KWOK, S ;
KELLOGG, DE ;
MCKINNEY, N ;
SPASIC, D ;
GODA, L ;
LEVENSON, C ;
SNINSKY, JJ .
NUCLEIC ACIDS RESEARCH, 1990, 18 (04) :999-1005
[10]   IDENTIFICATION OF HUMAN-IMMUNODEFICIENCY-VIRUS SEQUENCES BY USING INVITRO ENZYMATIC AMPLIFICATION AND OLIGOMER CLEAVAGE DETECTION [J].
KWOK, S ;
MACK, DH ;
MULLIS, KB ;
POIESZ, B ;
EHRLICH, G ;
BLAIR, D ;
FRIEDMANKIEN, A ;
SNINSKY, JJ .
JOURNAL OF VIROLOGY, 1987, 61 (05) :1690-1694