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目的监测市售贝壳类海产品中副溶血性弧菌的带菌状况、了解副溶血性弧菌的血清型别分布、耐药性特点及基因型别特征。方法随机采集15份汉沽海鲜市场麻蚶,按照GB4789.7-2013《食品微生物学检验副溶血性弧菌检验》进行分离鉴定,荧光定量PCR对耐热直接溶血素(tdh)和相关溶血素(trh)进行了毒力基因检测,K-B纸片扩散法对分离的菌株用15种抗生素进行耐药性监测,玻片凝集法进行血清分型,脉冲场凝胶电泳(PFGE)用BioNumerics软件,进行聚类分析。结果 15份麻蚶样品分离出副溶血性弧菌108株,毒力基因检测tdh和trh均为阴性。耐药性监测,79株(79/108)呈现单一耐药、22株(22/108)双重耐药、4株(4/108)为多重耐药,分别占73.15%,,20.37%,和3.7%。血清分型O群3个,其中O1群(78株)、O5群(24株)和未定型6株;K抗原定型12个,72株,未定型2个36株。PFGE分出102个不同的带型。结论汉沽区市售麻蚶中副溶血性弧菌检出率高,但菌株均未携带毒力基因。耐药性监测,97.22%的菌株至少对单一抗生素耐药,血清型别分布呈多样性,PFGE图谱呈多态性。
Objective To monitor the carriage of Vibrio parahaemolyticus in commercially available shellfish products and to understand the serotype distribution, drug resistance and genotype characteristics of Vibrio parahaemolyticus. Methods Fifteen randomly collected samples from Hangu seafood market were isolated and identified according to GB4789.7-2013 “Microbiological test of Vibrio parahaemolyticus”. Fluorescent quantitative PCR was used to detect the contents of heat-resistant direct hemolysin (tdh) and related hemolysin trh) were used to detect the virulence genes. KB strains were used to monitor the drug resistance of the isolated strains with 15 antibiotics. Serotypes were obtained by slide agglutination. Pulsed field gel electrophoresis (PFGE) was performed using BioNumerics software Cluster analysis. Results 108 strains of Vibrio parahaemolyticus were isolated from 15 samples of Mudanjiang and virulence genes were negative for tdh and trh. Drug resistance monitoring showed that 79 (79/108) showed single drug resistance, 22 (22/108) were double resistant and 4 (4/108) were multi-drug resistant, accounting for 73.15%, 20.37% 3.7%. There were 3 O serogroups, of which O1 group (78 strains), O5 group (24 strains) and unconfined type 6 strains. There were 12 Kk antigenotypes, 72 Kinds and 2 Undetermined strains. PFGE separated 102 different bands. Conclusion The detection rate of Vibrio parahaemolyticus was higher in Hangu district, but none of the strains carried the virulence genes. Drug resistance monitoring showed that 97.22% of the isolates were at least resistant to single antibiotics. The serotypes of the isolates showed diversity and the polymorphism of PFGE was polymorphic.