宁波地区腹泻患者副溶血弧菌毒力基因及耐药性分析

doi:10.3969/j.issn.1673-8640.2019.02.009

摘要/Abstract

摘要：

目的分析宁波地区腹泻患者副溶血弧菌毒力基因、Ⅲ型分泌系统（T3SS）及其耐药情况。方法收集分离自腹泻患者粪便样本的副溶血弧菌49株,采用聚合酶链反应（PCR）检测其毒力基因tdh、trh和T3SS1、T3SS2,采用纸片扩散法检测分离的菌株对17种常用抗菌药物的敏感性。结果 49株副溶血弧菌中有48株tdh阳性,1株trh阳性,47株tdh（+）trh（-）,1株tdh（+）trh（+）,1株tdh（-）trh（-）;49株副溶血弧菌均携带T3SS1基因,47株携带T3SS2α基因,1株携带T3SS2β基因。副溶血弧菌对氨苄西林的耐药率最高（75.5%）,对三代头孢、四环素类抗菌药物的敏感性较高,对头孢吡肟、左氧氟沙星和碳氢酶烯类抗菌药物（亚胺培南、美洛培南）的敏感性为100%。结论宁波地区腹泻患者副溶血弧菌毒力基因携带率高,对常见抗菌药物较敏感,应及时监测其耐药率变化,指导临床合理用药。

关键词: 副溶血弧菌, 腹泻, 毒力基因, 耐药性

Abstract:

Objective To investigate the virulence genes,type Ⅲ secretion system（T3SS） and drug resistance of Vibrio parahaemolyticus isolated from patients with diarrhea in Ningbo. Methods A total of 49 isolates of Vibrio parahaemolyticus were isolated from feces specimens of patients with diarrhea. The virulence genes,tdh,trh and T3SS1,T3SS2,were determined by polymerase chain reaction（PCR）. The drug susceptibility of 17 kinds of antibiotics were determined by disk diffusion assay. Results Among the 49 isolates of Vibrio parahaemolyticus,48 isolates carried tdh. There was 1 isolate carrying trh,47 isolates showed tdh（+）trh（-）,1 isolate showed tdh（+）trh（+）,and 1 isolate showed tdh（-）trh（-）. The 49 isolates carried T3SS1 gene,47 isolates carried T3SS2α gene,and 1 isolate carried T3SS2β gene. The drug resistance rate to ampicillin was 75.5%. The drug susceptibility rates were high to the 3rd generation cephalosporins and aminoglycoside tetracyclines. All the isolates were sensitive to cefepimes,levofloxacins and carbapenems（imipenem and meropenem）,and the drug susceptibility rates were 100%. Conclusions The carrying rates of virulence genes of Vibrio parahaemolyticus isolated from patients with diarrhea in Ningbo are high. The drug susceptibility rates are high to common antibiotics,and it should also pay attention to monitoring drug resistance rate and guiding rational antibiotic use in clinic.

Key words: Vibrio parahaemolyticus, Diarrhea, Virulence gene, Drug resistance

中图分类号:

R378.3

谢红意, 叶鸿雁, 周方满, 施露梅, 葛婷婷, 乐旭波. 宁波地区腹泻患者副溶血弧菌毒力基因及耐药性分析[J]. 检验医学, 2019, 34(2): 136-139.

XIE Hongyi, YE Hongyan, ZHOU Fangman, SHI Lumei, GE Tingting, LE Xubo. Virulence genes and drug resistance of Vibrio parahaemolyticus isolated from patients with diarrhea in Ningbo[J]. Laboratory Medicine, 2019, 34(2): 136-139.

图/表 3

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基因名称	序列（5'~3'）	长度（bp）	反应条件
tlh	AAAGCGGATTATGCAGAAGCACTG	450	94 ℃ 5 min; 94 ℃ l min,58 ℃ l min,72 ℃ l min, 30个循环;72 ℃ 10 min
	GCTACTTTCTAGCATTTTCTCTGC
trh	TTGGCTTCGATATTTTCAGTATCT	500	95 ℃ 5 min; 95 ℃ l min,54 ℃ l min,72 ℃ l min, 30个循环;72 ℃ 10 min
	CATAACAAACATATGCCCATTTCCG
tdh	GTAAAGGTCTCTGACTTTTGGAC	269	94 ℃ 5 min; 94 ℃ l min,58 ℃ l min,72 ℃ l min, 30个循环;72 ℃ 10 min
	TGGAATAGAACCTTCATCTTCACC

基因名称	序列（5'~3'）	长度（bp）	反应条件
tlh	AAAGCGGATTATGCAGAAGCACTG	450	94 ℃ 5 min; 94 ℃ l min,58 ℃ l min,72 ℃ l min, 30个循环;72 ℃ 10 min
	GCTACTTTCTAGCATTTTCTCTGC
trh	TTGGCTTCGATATTTTCAGTATCT	500	95 ℃ 5 min; 95 ℃ l min,54 ℃ l min,72 ℃ l min, 30个循环;72 ℃ 10 min
	CATAACAAACATATGCCCATTTCCG
tdh	GTAAAGGTCTCTGACTTTTGGAC	269	94 ℃ 5 min; 94 ℃ l min,58 ℃ l min,72 ℃ l min, 30个循环;72 ℃ 10 min
	TGGAATAGAACCTTCATCTTCACC

抗菌药物	敏感率	耐药率
头孢吡肟	100.0	0.0
左氧氟沙星	100.0	0.0
亚胺培南	100.0	0.0
美洛培南	100.0	0.0
哌拉西林-他唑巴坦	100.0	0.0
头孢他啶	97.8	0.0
头孢噻肟	93.9	0.0
四环素	91.8	0.0
氯霉素	89.8	1.6
庆大霉素	80.0	0.0
复方磺胺甲噁唑	75.5	1.5
头孢西丁	74.5	0.0
环丙沙星	74.5	2.7
头孢呋辛	26.5	20.4
阿米卡星	16.3	22.4
头孢唑啉	10.2	58.6
氨苄西林	10.2	75.5

抗菌药物	敏感率	耐药率
头孢吡肟	100.0	0.0
左氧氟沙星	100.0	0.0
亚胺培南	100.0	0.0
美洛培南	100.0	0.0
哌拉西林-他唑巴坦	100.0	0.0
头孢他啶	97.8	0.0
头孢噻肟	93.9	0.0
四环素	91.8	0.0
氯霉素	89.8	1.6
庆大霉素	80.0	0.0
复方磺胺甲噁唑	75.5	1.5
头孢西丁	74.5	0.0
环丙沙星	74.5	2.7
头孢呋辛	26.5	20.4
阿米卡星	16.3	22.4
头孢唑啉	10.2	58.6
氨苄西林	10.2	75.5