Beaulieu D, Bergeron M G, Roy P H
Département de Biochimie, Faculté des Sciences, Université Laval, Québec, Canada.
Mol Cell Probes. 1991 Feb;5(1):37-48. doi: 10.1016/0890-8508(91)90036-j.
Moraxella (Branhamella) catarrhalis is an aerobic Gram-negative diplococcus that is now recognized as a pathogen of the respiratory tract. Rapid and direct identification of this bacterium has become important to the clinical microbiology laboratory. Recently, rapid tests for the identification of Neisseria species and M. catarrhalis have been commercialized but they are primarily for Neisseriae; in these kits, M. catarrhalis is always identified presumptively. We have developed a DNA probe of chromosomal origin that is 100% specific for M. catarrhalis. The oligonucleotide probe was derived from a cloned Ase I chromosomal DNA fragment of M. catarrhalis that did not react with Haemophilus influenzae DNA in hybridization experiments. Three of the first 17 Ase I clones were selected randomly to be tested by colony hybridization against several different species that colonize the human respiratory tract. One of these three, pLQ121, has a 550 bp fragment inserted into pBR322 and was determined to be 100% specific to M. catarrhalis. This fragment was partially sequenced and a 30-mer oligonucleotide was synthesized from the sequence data. This probe was also shown to be 100% specific to the species.
卡他莫拉菌(以前称为卡他布兰汉菌)是一种需氧革兰氏阴性双球菌,现已被确认为呼吸道病原体。对这种细菌进行快速直接鉴定对临床微生物实验室来说已变得很重要。最近,用于鉴定奈瑟菌属和卡他莫拉菌的快速检测方法已商业化,但这些检测主要是针对奈瑟菌的;在这些试剂盒中,卡他莫拉菌总是被推定性鉴定。我们开发了一种源自染色体的DNA探针,它对卡他莫拉菌具有100%的特异性。该寡核苷酸探针源自卡他莫拉菌的一个克隆的Ase I染色体DNA片段,在杂交实验中它不与流感嗜血杆菌DNA发生反应。从最初的17个Ase I克隆中随机选择3个,通过菌落杂交对几种定殖于人类呼吸道的不同菌种进行检测。这3个克隆中的一个,即pLQ121,有一个550 bp的片段插入到pBR322中,经测定它对卡他莫拉菌具有100%的特异性。对该片段进行了部分测序,并根据序列数据合成了一个30聚体寡核苷酸。该探针也被证明对该菌种具有100%的特异性。