鱼源类志贺邻单胞菌的分子鉴定及耐药性分析

doi:10.13560/j.cnki.biotech.bull.1985.2019-0349

摘要/Abstract

摘要： 为对患病黄颡鱼分离的菌株（L-1）进行鉴定及耐药性分析,通过形态学观察、生理生化特性分析、16S rDNA基因和23S rRNA基因PCR扩增及系统进化树构建鉴定该菌,药敏试验和耐药基因检测确定菌株耐药性。结果表明：菌株L-1为革兰氏阴性菌,呈短杆状。明胶液化、七叶灵及阿拉伯糖等反应为阴性,精氨酸、色氨酸及葡萄糖等反应为阳性。16S rDNA基因扩增获得长度为1 461 bp的序列片段。BLAST比对结果显示,菌株L-1拼接序列与NCBI中类志贺邻单胞菌相似性为99%。在系统发育树中,菌株L-1与类志贺邻单胞菌亲缘关系最近。23S rRNA基因扩增,仅菌株L-1扩增出大小为280 bp的目的条带,其他菌株均为扩增出。综合判定菌株L-1为类志贺邻单胞菌。菌株L-1对头孢哌酮、庆大霉素等敏感,对四环素中度敏感,对苯唑西林、氨苄西林等耐药。菌株L-1中检测出Sul2、TEM、ant（3''）-Ⅰa和aac（6'）-Ⅰb共4种耐药基因,Sul1、Sul3、aph（3'）-Ⅱa和aac（3）-Ⅱa未检出。

关键词: 类志贺邻单胞菌, 鉴定, 耐药基因, 药敏试验

Abstract: To identify a strain（L-1）isolated from a sick Pelteobagrus fulvidraco and analyze its drug resistance,the morphology,physiological and biochemical analysis,16S rDNA gene and 23S rRNA gene amplification,and phylogenetic tree construction were carried out,and its drug resistance was determined by drug susceptibility test and drug-resistant genes test. The results showed that the strain L-1 was Gram-negative bacterium in short rob. The reactions to gelatin liquefaction,aesculin and arabinose and so on were negative,while that to arginine,tryptophan and glucose and so on were positive. The length of sequence fragment by 16S rDNA gene amplification was 1 461 bp. BLAST alignment showed that the similarity between the splicing sequence of L-1 and Plesiomonas shigellode in NCBI was 99%. In the phylogenetic tree of the strain L-1,the strain L-1 was the closest to P. shigellode. Only the target band with a size of 280 bp was amplified from the L-1 via 23S rRNA,while all other strains did not. Thus L-1 was comprehensively determined to be P. shigellode. Antimicrobial susceptibility test showed that L-1 was sensitive to cefoperazone,centamicin,etc.,and was moderate to tetracycline,and was resistant to benzocillin,ampicillin and other drugs. Drug-resistant gene Sul2,TEM,ant（3''）-Ⅰa and aac（6'）-Ⅰb in the strain L-1 were detected,while Sul1,Sul3,aph（3'）-Ⅱa and aac（3）-Ⅱa were not.

Key words: Plesiomonas shigellode, identification, drug resistance genes, antimicrobial susceptibility test

豆朋朋, 王利, 张桦, 郑姚. 鱼源类志贺邻单胞菌的分子鉴定及耐药性分析[J]. 生物技术通报, 2019, 35(11): 118-123.

DOU Peng-peng, WANG Li, ZHANG Hua, ZHENG Yao. Molecular Identification and Drug Resistance Analysis of Plesiomonas shigellode Isolated from Fish[J]. Biotechnology Bulletin, 2019, 35(11): 118-123.

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