FAST SOLID SUPPORT DETECTION OF HUMAN PAPILLOMAVIRUS IN IN-VITRO AMPLIFIED DNA USING A DNP ANTI DNP MONOCLONAL-ANTIBODY COUPLE

被引:12
作者
BACCARDLONGERE, M
ALPHABAZIN, B
CHYPRE, C
SAUVAIGO, S
TEOULE, R
BERNARD, P
SEIGNEURIN, JM
机构
[1] CIS BIOINT,SONDES MOLEC LAB,F-91192 GIF SUR YVETTE,FRANCE
[2] CEN,DRFMC,SESAM,RBIO,LSM,F-38041 GRENOBLE,FRANCE
[3] CHU GRENOBLE,SERV GYNECOL OBSTET,GRENOBLE 9,FRANCE
关键词
HUMAN PAPILLOMA VIRUS; POLYMERASE CHAIN REACTION; SOLID SUPPORT DETECTION; DNP;
D O I
10.1016/0166-0934(94)90014-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
In some anogenital lesions the detection of certain types of human papilloma virus, especially oncogenic types, is of interest. In a first step during a prospective study, we compared two methods for the detection of human papillomavirus (HPV) DNA in clinical samples: Southern blotting followed by hybridization with a cloned radioactive genomic probe and a classical polymerase chain reaction (PCR) followed by hybridization with a (32)p- labelled oligonucleotide probe. 118 biopsies and swabs were examined for HPV 6/11, 16, 18 and 33, 67 positive reactions were found by both methods, 5 positives only by PCR and 2 positives only by Southern blot for unidentified HPV. Patients with anogenital condylomas, dysplasias and carcinomas or asymptomatic patients were studied. Most high grade (II and III) dysplasias were associated with HPV 16 and HPV 18. Condylomata lesions and low grade dysplasia (grade I) were associated mostly with HPV 6/11, mixed type of HPV, less frequently with HPV 16 or HPV 18. As a second step a nested PCR coupled to solid support detection method was used as described by Sauvaigo et al. (1990) Nucleic Acids Res. 18, 3175-3183) to study a panel of 30 previously qualified different HPV DNA extracts. In this procedure the second round of PCR amplification involves biotinylated and dinitrophenylated labelled primers,allowing the capture of PCR amplified HPV DNA sequences on streptavidin coated tubes and its revelation. We describe an improvement of HPV DNA detection by means of single-step immunoenzymatic revelation involving anti-DNP monoclonal antibodies conjugated to horseradish peroxidase enzyme. A perfect correlation with the previous results was obtained. This solid support method allows a faster and easier HPV typing compared to methods using membrane transfer.
引用
收藏
页码:29 / 38
页数:10
相关论文
共 37 条
[21]  
Maniatis T., 1982, MOL CLONING
[22]   DOES HUMAN PAPILLOMAVIRUS CAUSE CERVICAL-CANCER - THE STATE OF THE EPIDEMIOLOGICAL EVIDENCE [J].
MUNOZ, N ;
BOSCH, X ;
KALDOR, JM .
BRITISH JOURNAL OF CANCER, 1988, 57 (01) :1-5
[23]   A TECHNIQUE FOR PREPARING DEFINED CONJUGATES OF HORSERADISH-PEROXIDASE AND IMMUNOGLOBULIN [J].
NILSSON, P ;
BERGQUIST, NR ;
GRUNDY, MS .
JOURNAL OF IMMUNOLOGICAL METHODS, 1981, 41 (01) :81-93
[24]  
OBANION MK, 1987, INTERVIROLOGY, V28, P114, DOI 10.1159/000150004
[25]  
ORTH G, 1988, B I PASTEUR, V86, P297
[26]  
REEVES WC, 1989, REV INFECT DIS, V11, P426
[27]   SYNTHESIS AND USE OF LABELED NUCLEOSIDE PHOSPHORAMIDITE BUILDING-BLOCKS BEARING A REPORTER GROUP - BIOTINYL, DINITROPHENYL, PYRENYL AND DANSYL [J].
ROGET, A ;
BAZIN, H ;
TEOULE, R .
NUCLEIC ACIDS RESEARCH, 1989, 17 (19) :7643-7651
[28]   PRIMER-DIRECTED ENZYMATIC AMPLIFICATION OF DNA WITH A THERMOSTABLE DNA-POLYMERASE [J].
SAIKI, RK ;
GELFAND, DH ;
STOFFEL, S ;
SCHARF, SJ ;
HIGUCHI, R ;
HORN, GT ;
MULLIS, KB ;
ERLICH, HA .
SCIENCE, 1988, 239 (4839) :487-491
[29]   FAST SOLID SUPPORT DETECTION OF PCR AMPLIFIED VIRAL-DNA SEQUENCES USING RADIOIODINATED OR HAPTEN LABELED PRIMERS [J].
SAUVAIGO, S ;
FOUQUE, B ;
ROGET, A ;
LIVACHE, T ;
BAZIN, H ;
CHYPRE, C ;
TEOULE, R .
NUCLEIC ACIDS RESEARCH, 1990, 18 (11) :3175-3183
[30]   PAPILLOMAVIRUS INFECTION OF THE LOWER GENITAL-TRACT - DETECTION OF VIRAL-DNA IN GYNECOLOGICAL SWABS [J].
SCHNEIDER, A ;
KRAUS, H ;
SCHUHMANN, R ;
GISSMANN, L .
INTERNATIONAL JOURNAL OF CANCER, 1985, 35 (04) :443-448