禽肉中弯曲菌的分离、PFGE和DGGE分型

Isolation of Campylobacter Strains in Poultry Products and Genotyping Identification of Strains by Means of PFGE and DGGE

随机选取某肉菜市场8份鸡肉和鸭肉样品中的弯曲菌(Campylobacter),分别采用Bolton和Preston肉汤增菌,然后以Skirrow、mCCDA选择分离培养和膜过滤的方法同时分离,运用脉冲场凝胶电泳(PFGE)和变性梯度凝胶电泳(DGGE)对分离出的菌株进行分子水平的分型和溯源.结果表明:共有6份样品污染了空肠弯曲菌(C.jejuni)或结肠弯曲菌(C.coli);Preston肉汤增菌、mCCDA选择分离培养法的分离效果最好;4份样品被同一种C.coli污染,2份样品分别污染了2种C.jejuni和3种C.coli;与PFGE相比,DGGE分型方法的灵敏度略低,但更快速、成本更低,且稳定、准确,可用于食品中弯曲菌的快速分型.

Eight poultry samples including chicken and duck were collected in a certain fresh market and were enriched in Bolton and Preston broths,followed by the inoculation on selective agar media（Skirrow and mCCDA） and by the membrane filtration.Then,the molecular types of the isolated Campylobacter strains were analyzed by means of pulse field gel electrophoresis（PFGE） and denaturing gradient gel electrophoresis（DGGE）,and C.jejuni or C.coli were isolated from six samples.The results show that the combination of Preston broth and mCCDA media is the optimal isolation method,that there are four samples contaminated by the same C.coli isolate and two samples contaminated respectively by 2 species of C.jejuni and 3 species of C.coli,and that,though DGGE genotyping is not as sensitive as PFGE genotyping,it is more suitable for the rapid genotyping of Campylobacter in food borne because it is of high speed,low cost as well as stable and accurate operation.