Cloeckaert A, Grayon M, Grepinet O
Laboratoire de Pathologie Infectieuse et Immunologie, Institut National de la Recherche Agronomique, 37380 Nouzilly, France.
Clin Diagn Lab Immunol. 2000 Sep;7(5):835-9. doi: 10.1128/CDLI.7.5.835-839.2000.
DNA polymorphism of the bp26 gene, coding for a diagnostic protein antigen for brucellosis, was assessed by PCR and restriction fragment length polymorphism analysis using primers to amplify the bp26 gene with its flanking regions. Surprisingly, whereas PCR performed on DNA of the reference strains of the six recognized Brucella species produced a product of the expected size (1,029 bp), PCR performed on DNA of three representative strains from marine mammals (from a seal, a dolphin, and a porpoise) produced a larger product, of about 1,900 bp. Nucleotide sequencing of the 1,900-bp PCR products revealed the presence of an insertion sequence, IS711, downstream of the bp26 gene and adjacent to a Bru-RS1 element previously described as being a hot spot for IS711 insertion. PCR performed on a large number of field strains from different geographic origins and from marine mammal isolates indicated that the occurrence of an IS711 element downstream of the bp26 gene was a feature specific to the marine mammal Brucella strains. Thus, this PCR assay is able to differentiate Brucella terrestrial isolates from marine mammal isolates and could be applied for diagnostic purposes.
通过聚合酶链反应(PCR)和限制性片段长度多态性分析,使用引物扩增布鲁氏菌病诊断蛋白抗原bp26基因及其侧翼区域,对编码该抗原的bp26基因的DNA多态性进行了评估。令人惊讶的是,虽然对六种公认的布鲁氏菌参考菌株的DNA进行PCR产生了预期大小(1029bp)的产物,但对来自海洋哺乳动物(海豹、海豚和鼠海豚)的三种代表性菌株的DNA进行PCR却产生了更大的产物,约1900bp。对1900bp PCR产物的核苷酸测序显示,在bp26基因下游存在一个插入序列IS711,且与先前被描述为IS711插入热点的Bru-RS1元件相邻。对来自不同地理来源的大量现场菌株以及海洋哺乳动物分离株进行PCR检测表明,bp26基因下游存在IS711元件是海洋哺乳动物布鲁氏菌菌株特有的特征。因此,这种PCR检测方法能够区分陆地布鲁氏菌分离株和海洋哺乳动物分离株,可用于诊断目的。