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目的对2起食物中毒中不同血清型的副溶血性弧菌进行基因特征分析。方法第1起食物中毒发生在2010年5月17日深圳市某食堂,感染人数20人左右,采集到病人肛拭子9份,可疑食品3份,每份样本挑取20个可疑菌落;第2起食物中毒发生在2010年7月1日深圳市某配餐中心,感染人数10人左右,采集到病人肛拭子7份,可疑食品2份,涂抹样3份,每份样本挑取10个可疑菌落。对可疑菌落分别进行盐耐受试验、生化试验和血清学鉴定。从2起食物中毒的每份样本中挑取不同血清型进行tdh、trh、GS-PCR、orf 8基因检测和脉冲场凝胶电泳分析(PFGE)。结果第1起食物中毒分离到副溶血性弧菌180株,其中O3:K6 107株,O2:K28 70株,O4:K34、O3:K25、O4:K12各1株;第2起食物中毒分离到副溶血性弧菌80株,其中O3:K6 44株,O4:K8、O11:K36、O11:K19各10株,O1:K56 6株。O3:K6、O1:K56、O4:K8、O11:K36携带tdh基因,不携带trh基因,其他血清型(O2:K28,O4:K12,O3:K25,O4:K34,O11:K19)均不携带tdh和trh基因。O3:K6和O11:K36 2种血清型的GS-PCR和orf 8基因阳性,其他血清型阴性。PFGE结果显示第1起食物中毒中食品和病人肛拭分离的O2:K28(分别3、2株)的PFGE带型基本一致,属高度相关菌株。第2起食物中毒中病人肛拭分离的4株O3:K6与卤鸡蛋盘涂抹样分离的1株O3:K6 PFGE带型一致。2起食物中毒的O3:K6带型基本一致。来自相同样本不同血清型的副溶血性弧菌的PFGE带型不一致。结论第1起食物中毒与食用污染了副溶血性弧菌的食品有关,第2起食物中毒与污染了副溶血性弧菌的卤鸡蛋盘有关。通过血清分型和分子生物学分析,可认为相同样本中分离的不同血清型副溶血性弧菌未发现遗传学证据,为遗传不相关菌株。
Objective To analyze the genetic characteristics of two strains of Vibrio parahaemolyticus in two kinds of food poisoning. Methods The first food poisoning occurred in a canteens in Shenzhen City on May 17, 2010, with a total number of 20 people infected. Nine samples of patients’ anal swabs and 3 suspicious samples were collected and 20 suspicious colonies were taken from each sample. 2 food poisoning occurred in July 1, 2010 in Shenzhen, a catering center, the number of people infected about 10, collected in patients with anal swab 7, 2 suspicious food, smear sample 3, each sample pick 10 Suspicious colonies. Susceptible colonies were salt tolerance test, biochemical tests and serological identification. Tdh, trh, GS-PCR, orf8 gene detection and pulsed-field gel electrophoresis (PFGE) were performed on each of the 2 food poisoning samples. Results 180 strains of Vibrio parahaemolyticus were isolated from the first food poisoning, among which 107 were O3: K6, 70 were O2: K28, 1 was O4: K34, O3: K25 and O4: K12 respectively. To Vibrio parahaemolyticus 80 strains, including O3: K6 44 strains, O4: K8, O11: K36, O11: K19 each 10 strains, O1: K56 6 strains. O3: K6, O1: K56, O4: K8, O11: K36 carry the tdh gene without trh gene, and none of the other serotypes (O2: K28, O4: K12, O3: K25, O4: K34, O11: K19) Carry tdh and trh genes. O3: K6 and O11: K36 two kinds of serotypes GS-PCR and orf 8 gene positive, the other serotypes negative. The results of PFGE showed that the PFGE patterns of O2: K28 (3,2 strains) separated from food and patient’s anal swab in the first food poisoning were basically the same, which were highly related strains. The 4 strains of O3: K6 separated from the anal swab in the second food poisoning were consistent with the one strain of O3: K6 PFGE band separated from the coated dish with brine eggs. 2 food poisoning O3: K6 band basically the same. Vibrio parahaemolyticus from different serotypes of the same sample have different PFGE patterns. Conclusions The first case of food poisoning was related to the consumption of food contaminated with Vibrio parahaemolyticus. The second case of food poisoning was related to the brine dish contaminated with Vibrio parahaemolyticus. By serotyping and molecular biology analysis, it can be concluded that there is no genetic evidence for the different serotypes of Vibrio parahaemolyticus isolated in the same sample, which is a genetically unrelated strain.