应用23S rRNA对禽波氏杆菌分离株的进化分析

Evolution analysis of 23S rRNA genes of Bordetella avium

通过PCR扩增分离保存的8株禽波氏杆菌、3株兔支气管败血波氏杆菌及1株猪支气管败血波氏杆菌菌株的23S rRNA基因的片段(710bp),分别克隆到载体pMD18-T后测序。利用BLAST工具进行同源性搜索;用DNAStar分析软件进行同源核苷酸序列的多重比较分析并构建禽波氏杆菌菌株的系统生物进化树。结果显示,8株禽波氏杆菌核苷酸序列之间同源性为99.2%～99.7%,与GenBank中收录的NC_010645株禽波氏杆菌核苷酸序列同源性为92.4%～92.5%,与兔支气管败血波氏杆菌和猪支气管败血波氏杆菌的核苷酸序列同源性均为98.5%～99.2%。国内分离的禽波氏杆菌菌株和支气管败血波氏杆菌菌株均与国外分离菌株在遗传基因上有一定的差异。结果表明,23S rRNA序列分析可以作为鉴定禽波氏杆菌的一种快速简便的方法。

To assess the evolution of Bordetella avium by 23S rRNA genes.The fragments（710 bp） of 23S rRNA genes were amplified by polymerase chain reaction（PCR） from eight Bordetella avium strains,three rabbit Bordetella bronchiseptica strains and one porcine Bordetella bronchiseptica strain.The PCR products were cloned into plasmid vector pMD-18T and then sequenced.The nucleotides homology analysis of the 23S rRNA gene was carried out using the BLAST.Multiple sequences alignments and the phylogenetic tree were performed using the DNAStar software.The strains of Bordetella avium and Bordetella bronchiseptica isolates from domestic and foreign countries have a certain differences in genetic.The homologies between eight Bordetella avium strains were 99.2%-99.7%,were 92.4%-92.5% with one strain in GenBank,and were 98.5%-99.2% with rabbit Bordetella bronchiseptica strain and porcine Bordetella bronchiseptica strain.The Bordetella avium strains and Bordetella bronchiseptica strains isolated in domestic have a certain degree of genetic difference with foreign countries strains.23S rRNA sequence analyses may be a reliable and rapid way for identifycation of Bordetella avium in diagnosis.