Detection and analysis of 5 strains of Vibrio cholerae O1 with virulence gene from green turtles and aquaculture water
摘要: 目的 监测杭州市江干区水产品中O1群和O139群霍乱弧菌并对分离菌株进行病原学鉴定及分子分型。 方法 按《霍乱防治手册》第5版对51份样品进行霍乱弧菌分离与鉴定。运用改良Kirby-Bauer 纸片法和PCR检测5株霍乱弧菌对14种临床常用抗生素的敏感性及是否携带ctxA和tcpA毒力基因,并用脉冲场凝胶电泳(PFGE)进行分子分型。 结果 从51份样品中检出O1群霍乱弧菌5株,其中小川型4株,稻叶型1株。5株霍乱弧菌的ctxA和tcpA毒力基因结果均为阳性。5株菌株对11种抗生素敏感,对红霉素和利福平中介,稻叶型菌株对氨苄西林敏感,小川型菌株对其耐药。PFGE分为两型,两型间仅有一条带的差异。 结论 本次分离的5株O1群霍乱弧菌均携带ctxA和tcpA毒力基因,故有必要加强对水产品尤其是甲鱼的监测及管理。Abstract: Objective To detect Vibrio cholerae O1 or O139 from aquatic products in Jianggan district and identify the pathogen and molecular genotype of the strains isolated. Methods The isolation and identification of Vibrio cholerae from 51 samples were conducted according to Manual for Cholera Prevention and Control. Kirby-Bauer method was used to test the susceptibility of the isolated strains of Vibrio cholerae to 14 antibiotics, PCR was performed to detect the virulence gene-ctxA and tcpA, and PFGE was used for molecular typing. Results Five strains of Vibrio cholerae O1 were detected from the 51 samples, including 4 Ogawa strains and 1 Inaba strain. The result of PCR for virulence gene detection showed they all carried ctxA and tcpA. The Ogawa strains were sensitive to Ampicillin but the Inaba strain was resistant to it, the 5 strains were all sensitive to 11 antibiotics, and mesomeric to erythromycin and rifampicin. The Results of PFGE showed that the 5 strains could be divided into two types, since there were only one fragment difference between them, Conclusion All the 5 isolated strains of Vibrio cholerae carried virulence gene-ctxA and tcpA, The surveillance and management of aquatic products, especially green turtles, should be strengthened.
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