山西省从粪便中分离到产霍乱毒素的拟态弧菌

Isolation of Vibrio mimicus producing cholera toxin from stools in Shanxi province

目的从山西省霍乱样粪便中分离到拟态弧菌,并检测其是否携带霍乱毒素基因。方法采集患者粪便、患者家用自来水、金鱼和鱼缸水标本,按照WS289-2008和《霍乱防治手册》(第5版)方法 ,对标本进行增菌和选择性培养基的分离培养,疑似菌落以霍乱弧菌血清凝集试验检测,并用API20E系统鉴定菌种。同时,疑似菌落用PCR检测O1、O139群特异性和是否携带霍乱毒素基因。结果粪便标本经增菌后接种选择性培养基,在庆大琼脂和碱性琼脂上均有霍乱弧菌疑似菌落,但在TCBS琼脂上为绿色、透明、中等大小、光滑、湿润的菌落;血平板上可见灰白色、湿润菌落,有透明溶血环;染色镜检湿片暗视野下未见流星状运动细菌。氧化酶和粘丝实验阳性,霍乱弧菌O1、O139群血清凝集试验阴性。菌株经API20E鉴定为拟态弧菌(93.5%可能性)。环境标本中未分离和检测到拟态弧菌。PCR检测该拟态弧菌中携带霍乱毒素基因,而O1、O139群特异性为阴性。结论从急性霍乱样腹泻粪便中分离到1株拟态弧菌,对于临床类似霍乱样症状的病例标本,当病原的霍乱弧菌血清凝集为阴性时,应进一步做霍乱毒素的检测。

Objective To isolate Vibrio mimicus （V.mimicus） from stool sample of cholera-like case,and detect whether it carried cholera toxin gene or not.Methods A diarrhea case with cholera-like symptom was investigated in Shanxi province.The stool and environmental samples were collected,cultured and isolated according to WS 289-2008 and cholera prevention and control manual （5th edition）,suspected colonies were detected with serum agglutination test of Vibrio cholerae （V.cholerae）,and API20E system was used to identify the bacteria.Meanwhile,the O1,O139 V.cholerae-specificity and cholera toxin gene of suspected colonies were detected by PCR.Results Stool samples were inoculated with selective medium,suspected colonies of V.cholerae were found in both gentamicin and alkaline agar,but in TCBS agar,green,transparent,medium size,smooth,moist colonies were observed;and in blood agar,gray,moist colonies and clear zone of hemolysis could be seen;After staining,no meteor-like movement of bacteria was observed on wet film under dark field.Oxidase and sticky silk test were positive,The serum agglutination test to detect V.cholerae O1,O139 and O141 were negative.The strain was identified as V.mimicus （93.5% possibility） with API20E.No V.mimicus was detected in environmental samples.PCR detection indicated the V.mimicus strain carried cholera toxin genes,but without V.cholerae O1,O139-specificity.Conclusion For specimens from cases of cholera like illness,when serum agglutination test of V.cholerae was negative,further detection of cholera toxin should be taken.