摘要
目的建立简便、快速、准确、特异的香港海鸥菌实验室诊断技术。方法采集淡水鱼类、胃肠炎腹泻患者新鲜粪样,接种于改良头孢哌酮MacConkey(CMA)培养基,对典型菌落分别进行氧化酶、触酶、三糖铁(TSI)3项筛选试验和常规生化鉴定,凡氧化酶、触酶试验阳性以及三糖铁(TSI)试验阴性的菌株,采用特异性PCR检测确诊;通用引物扩增16SrRNA并与香港海鸥菌HKU1株进行同源性分析。结果在92份淡水鱼样品和211份胃肠炎腹泻患者粪样中,共检出香港海鸥菌11株和1株,阳性率分别为11.96%(11/92)和0.48%(1/211)。筛选与确诊试验和常规生化试验的检测结果一致,符合率为100%(12/12)。分离的香港海鸥菌16SrRNA序列与HKU1株同源性在99.5%～100%之间,仅差1～2个碱基。结论本文建立的筛选与确诊试验适用于香港海鸥菌的诊断。
Objective To establish a set of methods for the simple,sharp,accurate,specific of Laribacter hongkongensis laboratory diagnosis.Methods Fecal swabs were freshly collected from the freshwater fishes and outpatients with diarrhea.Samples were grown on modified cefoperazone MacConkey agar（CMA）.Characteristic colonies on the selective medium were collected and tested with conventional biochemical tests.Samples showing positive results on the oxidase and catalase tests,negative on the triple sugar iron（TSI） agar were selected and further identified by PCR.16S rRNA was amplified by and compared with Laribacter hongkongensis strain HKU1.Results According to biochemical screening and PCR diagnosis test,12 strains of Laribacter hongkongensis were isolated from the 92 samples of freshwater fish and 211 cases of outpatients with diarrhea.The positive rate were 11.96%（11 /92）and 0.48%（1 /211）,respectively.The biochemical screening and PCR diagnosis results were all consistent with conventional biochemical identification.The sequence of 16S rRNA isolated from Laribacter hongkongensis was only 1 to 2 base pair different from that of the HKU1 strain,and the homology was 99.5% ～ 100.0%.Conclusion Protocol for the testing of Laribacter hongkongensis were established.PCR is a useful method in the identification of Laribacter hongkongensis.
出处
《热带医学杂志》
CAS
2010年第6期675-677,680,共4页
Journal of Tropical Medicine
基金
肇庆市科技创新计划项目(No.2009E1826)
关键词
香港海鸥菌
淡水鱼
胃肠炎
PCR
筛选
Laribacter hongkongensis
freshwater frish
gastroenteritis
PCR
screening
