两株水源性铜绿假单胞菌的分离与鉴定  

Isolation and Identification of Two Water-borne Pseudomonas Aeruginosa Strains

作　　者：梁涛波 龙慧[2] 占忠旭 莫逆 陈波 于帆 吴鑫 LIANG Taobo;LONG Hui;ZHAN Zhongxu;MO Ni;CHEN Bo;YU Fang;WU Xin(Jiangxi Institute of Food Control,Nanchang 330001,China;Nanchang Center for Disease Control and Prevention,Nanchang 330038,China;Jiangxi Runtian Industrial Co.,Ltd.,Nanchang 330029,China)

机构地区：[1]江西省食品检验检测研究院,江西南昌330001 [2]南昌市疾病预防控制中心,江西南昌330038 [3]江西润田实业股份有限公司,江西南昌330029

出　　处：《现代食品》2021年第17期206-212,共7页Modern Food

基　　金：2020年国家市场监督管理总局科技计划项目(2020MK057);2020年江西省科技厅重点研发项目(20203BBG73064)。

摘　　要：目的:分析桶装水样品中分离的两株产绿脓菌素铜绿假单胞菌(Pseudomonas aeruginosa,PA)生物学特性。方法:两株分离株在CN琼脂平板上划线培养48 h获取单菌落,使用VITEK进行生化鉴定和药敏分析,采用荧光定量PCR检测毒力基因和溶血基因,并使用MEGA-X进行同源性分析。结果:两株铜绿假单胞菌分离株菌落均呈现浅绿色,紫外灯照射显示有较强的绿色荧光,革兰氏染色结果显示两株铜绿假单胞菌分离株均为革兰氏阴性棒状杆菌,菌体较为光滑,大小适中。VITEK生化鉴定结果表明两株分离株为铜绿假单胞菌,比对发现两株铜绿假单胞菌生化分析结果存在一定的差异,药敏分析结果显示两株分离株均为多重耐药菌,并且相对于标准参考株ATCC 27853耐药信息更为广泛。铜绿假单胞菌分离株携带溶血基因、T2SS分泌系统、T3SS分泌系统相关的6种毒力基因。16s rRNA测序同源性比对以及系统进化树分析发现,两株分离株的16s rRNA序列与目前NCBI报道的铜绿假单胞菌具有较高同源性。结论:本研究通过多种方法进行验证和检验,更加了解两株水源性铜绿假单胞菌分离株生物学特性。Objective:To analyze the biological characteristics of two strains of Pseudomonas aeruginosa isolated from a bottled water sample.Methods:P.aeruginosa isolates were streaked on CN agar plates for 48 hours to obtain a single colony.VITEK was used for biochemical identification and drug sensitivity analysis,and fluorescence quantitative PCR was used to detect virulence genes and hemolytic genes,and MEGA-X was used for homology analysis.Results:The colonies of the P.aeruginosa isolates were light green,and UV light irradiation showed strong green fluorescence.Gram staining results showed that the P.aeruginosa isolates were both Gram-negative coryneform bacilli,with smoother cells and moderate size.The biochemical identification results of VITEK showed that the two isolates were P.aeruginosa.The comparison found that the biochemical analysis results of the two P.aeruginosa revealed certain differences.The results of drug sensitivity analysis showed that the two isolates were multi-drug resistant bacteria,and compared with the standard reference strain ATCC 27853 drug resistance information is more extensive.P.aeruginosa isolates carry six virulence genes related to hemolysis gene,T2SS secretion system and T3SS secretion system.The 16s rRNA sequencing homology comparison and phylogenetic tree analysis found that the 16s rRNA sequences of the two isolate strains have high homology with the current P. aeruginosa reported by NCBI. Conclusion: In this study, a variety of methods were used to verify and test, and to gain a better understanding of the biological characteristics of the two P. aeruginosa isolates.

关 键 词：铜绿假单胞菌 绿脓菌素 生化鉴定 药敏测试 同源性分析 

分 类 号：R123.1[医药卫生—环境卫生学]