香港海鸥形菌分型方法建立与病原学分布调查

Survey on Establishment of Molecular Subtyping Method and Pathogenic Distribution of Laribacter hongkongensis

目的通过对深圳市感染性腹泻患者标本与外环境水产品标本的检测,掌握深圳市香港海鸥形菌病原学特征及分子分型。方法用传统方法和荧光PCR法同时检测香港海鸥形菌,阳性菌株用K-B法做药敏试验和PFGE分子分型。结果在329份淡水鱼中共检出7份阳性标本,579份腹泻病人标本未检出。该菌对临床常用的24种抗生素耐药性检测结果显示,对氨苄西林、头孢他啶、头孢噻吩、头孢噻腭、头孢哌酮、头孢曲松耐药,头孢西丁为中介,其余均为敏感。7株香港海鸥形菌生化反应阳性的,荧光PCR结果均为阳性。PFGE分子分型表明7株菌不具有同源性。结论本文建立一套完整的对该菌的分离鉴定、基因诊断和PFGE分子分型方法;了解深圳地区主要流行菌型,预测深圳地区该菌的流行趋势,为应对食源性疾病突发事件和水产品安全问题做好技术储备。

Objective To investigate the pathogenic characteristics and molecular subtyping of Laribacter hongkongensis by detecting clinical samples collected from infectious diarrhea patients and aquatic product samples in Shenzhen. Methods All samples were detected by the traditional method and the developed real-time PCR assay.Drug-sensitive test with Kirby-Bauer method and PFGE molecular subtyping were carried out on the positive samples. Results Seven positive samples were detected from 329 freshwater fish samples, but no sample was detected in 579 clinical samples from diarrhea patients.Antimicrobial susceptibility of Laribacter hongkongensis to 24 kinds of commonly used antibiotics showed that Laribacter hongkongensis was resistant to AMP,CAZ,KF,CTX,SCF,and CTRX.FOX was intermediate,and the others were sensitive.The results tested by biochemical reactions and real-time PCR assay were in accordance with the traditional method.PFGE molecular subtyping showed that 7 positive samples did not have homology. Conclusions The method of isolation and identification,PCR detection,PFGE molecular subtyping for Laribacter hongkongensis was developed.It could provide detection methods for foodborne disease outbreaks of Laribacter hongkongensis and guarantee food safety.