Picken R N, Wang Z, Yang H L
ENZO Biochem, Inc., New York, NY 10013.
Mol Cell Probes. 1987 Sep;1(3):245-59. doi: 10.1016/0890-8508(87)90037-5.
Conventional procedures for isolating and identifying Campylobacter jejuni are cumbersome and time-consuming. A simpler approach would be to use DNA probes to identify these organisms. To obtain such probes we cloned chromosomal DNA from C. jejuni into the lambda replacement vector EMBL 4. Recombinant phages were screened for C. jejuni-specific inserts by DNA hybridization using chromosomal DNA from either C. jejuni or C. coli which had been radioactively labelled with 32P. By this means, recombinant phages were identified which hybridized to C. jejuni but not to C. coli DNA. These phages were then subjected to further screening using DNA from other Campylobacter species. Three DNA fragments were identified which hybridized to DNA from eight ATCC (American Type Culture Collection) strains of C. jejuni but not to DNA from C. coli, C. laridis, C. fetus or a variety of other bacterial species. These DNA fragments are suitable for use as specific probes in DNA-based diagnostic tests for C. jejuni infections.
空肠弯曲菌的传统分离和鉴定程序繁琐且耗时。一种更简单的方法是使用DNA探针来鉴定这些微生物。为了获得此类探针，我们将空肠弯曲菌的染色体DNA克隆到λ置换载体EMBL 4中。使用已用32P放射性标记的空肠弯曲菌或大肠弯曲菌的染色体DNA，通过DNA杂交筛选重组噬菌体，以寻找空肠弯曲菌特异性插入片段。通过这种方法，鉴定出了与空肠弯曲菌DNA杂交但不与大肠弯曲菌DNA杂交的重组噬菌体。然后使用来自其他弯曲菌属物种的DNA对这些噬菌体进行进一步筛选。鉴定出了三个DNA片段，它们与八株美国典型培养物保藏中心（ATCC）的空肠弯曲菌菌株的DNA杂交，但不与大肠弯曲菌、拉氏弯曲菌、胎儿弯曲菌或其他多种细菌物种的DNA杂交。这些DNA片段适合用作基于DNA的空肠弯曲菌感染诊断测试中的特异性探针。
