DETECTION OF ENTEROTOXIGENIC ESCHERICHIA-COLI IN FECAL SPECIMENS BY ACETYLAMINOFLUORENE-LABELED DNA PROBES

被引：5

作者：

BEGAUD, E ^{[1
]}

GERMANI, Y ^{[1
]}

机构：

[1] INST PASTEUR NOUVELLE CALEDONIE, BACTERIES ENTEROPATHOGENES LAB, BP 61, NOUMEA, NEW CALEDONIA

来源：

RESEARCH IN MICROBIOLOGY | 1992年 / 143卷 / 03期

关键词：

ACETYLAMINOFLUORENE; ENTEROTOXIN; ESCHERICHIA-COLI; DETECTION; FECES; AAF-DNA PROBES; ETEC;

D O I：

10.1016/0923-2508(92)90023-H

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

The present study describes acetylaminofluorene(AAF)-modified DNA probes for the identification of heat-labile (LT) and heat-stable (porcine STp and human STh) toxins from enterotoxigenic Escherichia coli (ETEC). AAF probes were compared with established biotinylated probes and bioassays. Ultracentrifugation was not necessary in the preparation of AAF-labelled probes, and the procedures, i.e. chemical modification of probes, hybridization and immunodetection steps, were optimized to detect ETEC by colony hybridization or direct dot blot techniques. The method combines chemical labelling (covalent attachment of AAF group to guanosine is achieved by treatment of DNA with N-acetoxy-2-acetylaminofluorene) and detection of the hybridized target DNA by means of anti-AAF monoclonal antibodies and alkaline phosphatase-labelled second antibodies.

引用

页码：315 / 325

页数：11

共 31 条

[1] DETECTION OF ENTEROTOXIGENIC ESCHERICHIA-COLI BY DOT BLOT HYBRIDIZATION WITH BIOTINYLATED DNA PROBES [J].

BIALKOWSKAHOBRZANSKA, H .

JOURNAL OF CLINICAL MICROBIOLOGY, 1987, 25 (02) :338-343

[2]

BIRNBOIM HC, 1979, NUCLEIC ACIDS RES, V7, P1513

[3] IDENTIFICATION AND CLASSIFICATION OF CAMPYLOBACTER STRAINS BY USING NONRADIOACTIVE DNA PROBES [J].

CHEVRIER, D ;

LARZUL, D ;

MEGRAUD, F ;

GUESDON, JL .

JOURNAL OF CLINICAL MICROBIOLOGY, 1989, 27 (02) :321-326

[4] CISTRONS ENCODING ESCHERICHIA-COLI HEAT-LABILE TOXIN [J].

DALLAS, WS ;

GILL, DM ;

FALKOW, S .

JOURNAL OF BACTERIOLOGY, 1979, 139 (03) :850-858

[5] PHYSICAL BASIS OF CHEMICAL CARCINOGENESIS BY N-2-FLUORENYLACETAMIDE DERIVATIVES AND ANALOGS [J].

FUCHS, R ;

DAUNE, M .

FEBS LETTERS, 1973, 34 (02) :295-298

[6]

GEOFFROY C, 1978, ANAL EAU, P35

[7] ACETYLAMINOFLUORENE-LABELED RIBOSOMAL-RNA FOR USE IN MOLECULAR EPIDEMIOLOGY AND TAXONOMY [J].

GRIMONT, F ;

CHEVRIER, D ;

GRIMONT, PAD ;

LEFEVRE, M ;

GUESDON, JL .

RESEARCH IN MICROBIOLOGY, 1989, 140 (07) :447-454

[8]

GUERRANT RL, 1985, PATHOGENESIS ESCHERI, P68

[9]

HAAS MJ, 1986, NUCLEIC ACIDS RES, V14, P3976

[10] SYNTHETIC OLIGODEOXYRIBONUCLEOTIDE PROBES FOR DETECTING HEAT-STABLE ENTEROTOXIN-PRODUCING ESCHERICHIA-COLI BY DNA COLONY HYBRIDIZATION [J].

HILL, WE ;

PAYNE, WL ;

ZON, G ;

MOSELEY, SL .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1985, 50 (05) :1187-1191

← 1 2 3 4 →