摘要
采用16SrRNA基因序列分析法对2015年7-9月广东省食品检验所桶装水专项抽检中筛查所得的81株铜绿假单胞菌（Pseudomonas aeruginosa）进行复核鉴定。菌株经纯化培养后提取总DNA,采用细菌16SrRNA通用引物进行16SrRNA基因序列扩增,PCR扩增产物经2%琼脂糖凝胶电泳后,进行序列测定,序列经人工校对后用Clustal X进行比对分析,最后用MEGA5.1软件构建系统发育树。系统发育分析结果 表明：81株铜绿假单胞菌与原鉴定结果一致。其中,编号24-3-QY、100-5-JM、106-3-JM菌株形成一个分支,28-1-WD单独为一支,其余77株野生菌和铜绿假单胞菌标准菌株ATCC27853聚为一群。该研究是2015年5月24日中国开始实施GB 19298—2014《食品安全国家标准包装饮用水》以来,广东省首次对水源性铜绿假单胞菌进行的研究,为下一步菌种污染朔源等研究提供依据。
81 Pseudomonas aeruginosa from Guangdong Provincial Institute of Food Inspection were identified by 16S rRNA sequence analysis. The DNA was isolated and the sequences of 16S rRNA gene were amplified by PCR with the bacterium universal primers, and then the PCR products were sequenced after 2 % agarose gel electrophoresis. Moreover, the corrected sequences were aligned with Clustal X and the phylogenetic tree was constructed by MEGA5.1. Consequently, the identified results of the 81 strains confirmed their original identification before. On the phylogenetic tree, No. 24-3-QY strain formed a separate branch with No. 100-5-JM strain and No.106-3-JM strain, No.2 8-1-DW formed one branch and the other 77 strains formed a separate branch with P. aeruginosa ATCC 27853. This was the first research about waterborne P. aeruginosa in Guangdong Province when China began to implement GB 19298-- 2014 ＂National food safety standards of packaged drinking water＂ since May 24, 2015. The waterborne P. aeruginosa culture collection in Guangdong was preliminarily established basing on these strains. These data will provide a powerful tool for effectively tracing source of P. aeruginosa and controlling the water contamination in future.
出处
《食品与机械》
CSCD
北大核心
2016年第11期21-24,89,共5页
Food and Machinery
