Gebhart C J, Murtaugh M P, Lin G F, Ward G E
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Minnesota, St. Paul 55108.
Vet Microbiol. 1990 Sep;24(3-4):367-79. doi: 10.1016/0378-1135(90)90184-w.
Cloned, chromosomal DNA probes from porcine isolates of Campylobacter hyointestinalis and C. mucosalis were developed for the detection and identification of these putative swine enteric pathogens. High molecular weight chromosomal DNA from each species was used to construct genomic libraries in plasmids. Recombinants were selected which hybridized strongly to the homologous organism, but not to any other species of Campylobacter. Species-specific recombinants were labeled with phosphorus-32 and tested for sensitivity by dot blot hybridization to various dilutions of DNA and bacteria from each swine species, including C. hyointestinalis, C. mucosalis, C. coli and C. jejuni. Specificity was tested by hybridizing these probes against various strains of C. hyointestinalis or C. mucosalis, and against reference strains of all other described Campylobacter species. A C. hyointestinalis-specific probe and a C. mucosalis-specific probe were identified which were capable of detecting 1 ng of DNA or 10(4) cfu by bacterial spot blotting on nylon membranes. These probes hybridized to intestinal mucosal scrapings containing C. hyointestinalis and C. mucosalis obtained from pigs with proliferative enteritis, but not to material from normal pigs. Thus, cloned, chromosomal DNA probes may be useful in the detection and identification of bacteria involved in swine proliferative enteritis.
构建了来自猪源肠道弯曲杆菌和黏膜弯曲杆菌分离株的克隆染色体DNA探针，用于检测和鉴定这些假定的猪肠道病原体。利用每个物种的高分子量染色体DNA构建质粒基因组文库。筛选出与同源生物体强烈杂交，但不与任何其他弯曲杆菌物种杂交的重组体。用磷-32标记物种特异性重组体，并通过斑点印迹杂交检测其对来自每个猪种（包括肠道弯曲杆菌、黏膜弯曲杆菌、大肠弯曲杆菌和空肠弯曲杆菌）的各种稀释度的DNA和细菌的敏感性。通过将这些探针与各种肠道弯曲杆菌或黏膜弯曲杆菌菌株以及所有其他已描述的弯曲杆菌物种的参考菌株杂交来测试特异性。鉴定出一种肠道弯曲杆菌特异性探针和一种黏膜弯曲杆菌特异性探针，它们能够通过在尼龙膜上进行细菌斑点印迹检测到1 ng DNA或10(4) cfu。这些探针与从患有增生性肠炎的猪中获得的含有肠道弯曲杆菌和黏膜弯曲杆菌的肠黏膜刮片杂交，但不与正常猪的材料杂交。因此，克隆的染色体DNA探针可能有助于检测和鉴定参与猪增生性肠炎的细菌。
