A SPECIFIC DNA PROBE FOR DETECTING MYCOPLASMA-HYOPNEUMONIAE IN EXPERIMENTALLY INFECTED PIGLETS

被引:14
作者
ABIVEN, P
BLANCHARD, B
SAILLARD, C
KOBISCH, M
BOVE, JM
机构
[1] INRA, BIOL CELLULAIRE & MOLEC LAB, F-33883 VILLENAVE DORNON, FRANCE
[2] UNIV BORDEAUX 2, F-33883 VILLENAVE DORNON, FRANCE
关键词
MYCOPLASMA-HYOPNEUMONIAE; DNA PROBE; SWINE ENZOOTIC PNEUMONIA; HYBRIDIZATION; DETECTION;
D O I
10.1016/0890-8508(92)90037-X
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Mycoplasma hyopneumoniae is the primary agent of swine enzootic pneumonia. Because of fastidious growth requirements and its serological cross-reactions with other porcine mycoplasmas, we developed a specific DNA probe for its detection. A partial genomic library of M. hyopneumoniae was constructed in plasmid pBR 322 using Hin dlll chromosomal fragments. The recombinant plasmids were screened by differential hybridization with M. flocculare and M. hyorhinis genomic DNA probes. One non-hybridizing recombinant plasmid was selected and its 1.65 kbp insert (designated l141) tested for specificity against genomic DNA from numerous mycoplasmas, other bacteria species and DNA from lung tissue of specific pathogen free (SPF) piglets. The 32P labelled l141 could detect specifically down to 400 pg of M. hyopneumoniae genomic DNA. To test the suitability of the l141 probe for the laboratory diagnosis of M. hyopneumoniae infections, we used clinical tracheobronchial specimens from piglets which were experimentally infected with M. hyopneumoniae. The results with hybridization on each specimen were compared to findings with an immunofluorescence test. Of the clinical specimen tested, there was agreement in the two tests of 63%. © 1992.
引用
收藏
页码:423 / 429
页数:7
相关论文
共 31 条
[1]   IDENTIFICATION OF MYCOPLASMA-HYOPNEUMONIAE WITH A DNA PROBE [J].
AHRENS, P ;
FRIIS, NF .
LETTERS IN APPLIED MICROBIOLOGY, 1991, 12 (06) :249-253
[2]  
AMANFU W, 1984, AM J VET RES, V45, P1349
[3]  
ARMSTRONG CH, 1987, ISRAEL J MED SCI, V23, P654
[4]  
BIRNBOIM HC, 1979, NUCLEIC ACIDS RES, V7, P1513
[5]   ELECTRON-MICROSCOPIC OBSERVATION OF THE RESPIRATORY-TRACT OF SPF PIGLETS INOCULATED WITH MYCOPLASMA-HYOPNEUMONIAE [J].
BLANCHARD, B ;
VENA, MM ;
CAVALIER, A ;
LELANNIC, J ;
GOURANTON, J ;
KOBISCH, M .
VETERINARY MICROBIOLOGY, 1992, 30 (04) :329-341
[6]   SPECIES-SPECIFIC ANTIGENS OF MYCOPLASMA-HYOPNEUMONIAE AND CROSS-REACTIONS WITH OTHER PORCINE MYCOPLASMAS [J].
BOLSKE, G ;
STRANDBERG, ML ;
BERGSTROM, K ;
JOHANSSON, KE .
CURRENT MICROBIOLOGY, 1987, 15 (04) :233-239
[7]   DETECTION AND DIFFERENTIATION OF THE MYCOPLASMALIKE ORGANISM ASSOCIATED WITH APPLE PROLIFERATION DISEASE USING CLONED DNA PROBES [J].
BONNET, F ;
SAILLARD, C ;
KOLLAR, A ;
SEEMULLER, E ;
BOVE, JM .
MOLECULAR PLANT-MICROBE INTERACTIONS, 1990, 3 (06) :438-443
[8]  
FALK K, 1991, ACTA VET SCAND, V32, P67
[9]  
FRIIS NF, 1972, ACTA VET SCAND, V13, P284
[10]  
FRIIS NF, 1975, NORD VET MED, V27, P337