四川地区鸡场产气荚膜梭菌的扩增片段长度多态性分析

Analysis of chicken Clostridium perfringens isolated from Sichuan province by amplified fragment length polymorphism

目的了解四川部分地区10个规模化鸡场产气荚膜梭菌的基因型及遗传学关系。方法采用EcoRI、MseI酶对34株不同地区鸡场的产气荚膜梭菌分离株进行扩增片段长度多态性(amplified fragment length polymorphism,AFLP)分析。结果AFLP能全部区别34株产气荚膜梭菌,辨别力指数为100%,其不同来源菌株呈现多态性分布。按Dice系数≥0.8,可分为19个AFLP基因亚型,AFLP-14为优势基因型,占总菌株数的20.59%。分析亚型分布情况表明,产气荚膜梭菌的流行特点与地区差异相关:不同鸡场产气荚膜梭菌的亚型差异明显;而同一鸡场的亚型较单一,以一种亚型为主,交叉有少数其他亚型。结论四川地区鸡场产气荚膜梭菌的基因型十分丰富,AFLP技术是其分子流行病学研究的有效手段。

To analyze the genetic relationship and predominate genotypes among Clostridium perfringens isolated from Chicken farm in Sichuan province.Of 34 C.perfringens strains were genotyped by amplified fragment length polymorphism（APLP） protocol using EcoR I and Mse I.lt was found that all strains were typeable by AFLP,and the discriminative index was 100%.In total 19 AFLP-genotype clusters were discriminated by 0.8 dice coefficient,and the AFLP-14 cluster was predominate,accounting to 20.59% of the total isolates.The analysis of genetic type and origin source revealed that differentiation correlated closely with the origins of the isolates.Isolates from different farms had different genetic types.The subtype of C.perfringen appeared to be simple within a single farm,and mixed with fewer other gene subtypes.It is evident that the genetic polymorphism of C.perfringens is diverse in chicken farm of Sichuan province,and amplified fragment length polymorphism is one of the effective means of molecular epidemiology.