采用PCR方法鉴别伴放线放线杆菌的6种血清型

Identification of Actinobacillus actinomycetemcomitans serotypes by polymerase chain reaction

目的:探索采用PCR的方法对伴放线放线杆菌的不同菌株进行血清型分类。方法:根据伴放线放线杆菌不同血清型特异性多糖抗原基因序列设计6对不同的寡核苷酸引物,用这6对引物分别对所选择伴放线放线杆菌6种不同的血清型菌株各3株,共18株,其中参考菌株6株,系ATCC29523(血清型a),ATCC43718(血清型b),ATCC33384(血清型c),IDH781(血清型d),IDH1705(血清型e)以及CU1000(血清型f),其余12个菌株均为临床分离株的DNA进行PCR扩增分析。结果:每一对引物均针对相应的血清型产生特异性单一条带PCR产物,产物大小分别为428bp(a),298bp(b),559bp(c),690bp(d),211bp(e),232bp(f)。全部18个菌株均能够被准确识别,无交叉反应。结论:PCR方法可以快速准确地鉴别伴放线放线杆菌目前已知的全部6种血清型。

Objective：To investigate a method to identify Actinobacillus serotypes by polymerase chain reaction. Methods： 18 Actinobacillus actinomycetemcomitans strains representing serotypes a to f were used in the test. 6 pairs of specific oligonucleotide primers for gene clusters involved in the biosynthesis of serotype specific polysaccharide antigens were designed. The primers were developed and evaluated in a genetic method of identifying serotypes of Actinobacillus actinomycetemcomitans strains by using a PCR assay. Results： Each pair of primers can specifically identify one serotype of Actinobacillus strains, No cross reaction was observed in all 18 strains. The PCR product sizes were as follows： 428 bp （serotype a）, 298 bp （serotype b）, 559 bp （serotype c）, 690 bp （serotype d）, 211 bp （serotype e） and 232 bp （serotype f）. Conclusion：PCR method may be useful to identify Actinobacillus actinomycetemcomitans serotypes rapidly and directly.