Savelkoul P H, de Groot L E, Boersma C, Livey I, Duggleby C J, van der Zeijst B A, Gaastra W
Department of Bacteriology, Faculty of Veterinary Medicine, University of Utrecht, The Netherlands.
Microb Pathog. 1993 Sep;15(3):207-15. doi: 10.1006/mpat.1993.1071.
A DNA fragment from Bordetella pertussis, encoding the fim2 fimbrial subunit gene with adjacent sequences, was used as a probe for the detection of homologous sequences in chromosomal DNA of Bordetella avium. A 1.8 kb Sa1I-PstI fragment from the genome of B. avium, which hybridized with the probe, was isolated and sequenced. No fimbrial subunit gene was located on the B. avium DNA fragment. Two regions could be distinguished in the sequence of the fragment. Region 1, which was 80% identical to the sequence upstream of the fim2 gene of B. pertussis and region 2, which had no identity with any known sequence. A 491 bp EagI DNA fragment (probe A) within region 1 and a 650 bp EagI DNA fragment (probe B) within region 2 were used as DNA probes on restriction endonuclease digests of chromosomal DNA from various bacterial species. This hybridization experiment showed that the region 2 sequence was specific for B. avium. A polymerase chain reaction (PCR) with specific primers within region 2 resulted in the amplification of a 500 bp DNA fragment with B. avium DNA only. This PCR is a useful method for the rapid detection of B. avium and appeared useful to discriminate B. avium from other Bordetella species and also from Alcaligenes faecalis.
来自百日咳博德特氏菌的一段编码fim2菌毛亚基基因及相邻序列的DNA片段，被用作探针来检测鸟博德特氏菌染色体DNA中的同源序列。从鸟博德特氏菌基因组中分离出一个与该探针杂交的1.8 kb Sa1I - PstI片段并进行测序。在鸟博德特氏菌DNA片段上未定位到菌毛亚基基因。该片段序列可区分出两个区域。区域1与百日咳博德特氏菌fim2基因上游序列有80%的同源性，区域2与任何已知序列均无同源性。区域1内的一个491 bp EagI DNA片段（探针A）和区域2内的一个650 bp EagI DNA片段（探针B）被用作针对各种细菌物种染色体DNA限制性内切酶消化产物的DNA探针。该杂交实验表明区域2序列对鸟博德特氏菌具有特异性。用区域2内的特异性引物进行聚合酶链反应（PCR），结果仅从鸟博德特氏菌DNA中扩增出一个500 bp的DNA片段。这种PCR是快速检测鸟博德特氏菌的一种有用方法，且似乎有助于将鸟博德特氏菌与其他博德特氏菌属物种以及粪产碱菌区分开来。
