实时荧光PCR法、胶体金法和培养法检测甲鱼中霍乱弧菌  

Detection of Vibrio Cholerae in Turtles by Real Time Polymerase Chain Reaction,Colloidal Gold Immunochromatographic Assay and Conventional Bacterial Culture

作　　者：颜淑妩[1] 李哲婷[1] 邓婵[1] 

机构地区：[1]湖南省衡阳市疾病预防控制中心,湖南衡阳421001

出　　处：《实用预防医学》2012年第6期925-927,共3页Practical Preventive Medicine

摘　　要：目的优化水产品甲鱼中霍乱弧菌的检测程序,提高甲鱼中霍乱弧菌检出率。方法用实时荧光PCR、常规细菌培养、胶体金法同时对甲鱼中霍乱弧菌进行检测,并用实时荧光PCR法检测标本中霍乱弧菌ctx基因。结果共检测185份甲鱼样品,其中实时荧光PCR法检出28份霍乱弧菌核酸阳性,阳性率为15.14%;6份ctx基因核酸阳性,阳性率21.43%(6/28)。常规细菌培养法分离出2株菌株,一株为O139群霍乱弧菌,一株为小川型霍乱弧菌,用实时荧光PCR检测这两株纯培养菌株或原始标本,霍乱弧菌ctx基因均为阴性;胶体金法未检出阳性标本。结论对于水产品标本,可先用实时荧光PCR法筛检霍乱弧菌,阳性标本再进行传统细菌分离培养,以提高霍乱弧菌菌株的检出率;同时阳性标本进行霍乱弧菌ctx基因核酸检测,如也为阳性,需提高警惕,加强流行病学上的预防控制措施,及时防范霍乱疫情的发生。Objective To optimize the detection procedure of Vibrio Cholerae（v.cholerae） and increase its detection rate in turtles.Methods The v.cholerae in turtles was detected by real time polymerase chain reaction（real time PCR）,conventional bacterial culture and colloidal gold immunochromatographic assay,and the ctx gene of the virus was detected by real time PCR.Results Real time PCR revealed that among 185 turtle samples,28 ones were positive with v.cholerae nucleic acid,with a positive rate of 15.14%（28/185）,and six samples were positive with ctx gene,with a positive rate of 21.43%（6/28）.Two strains of v.cholerae were isolated by conventional bacterial culture,Vibrio cholerae O139,and Vibrio cholerae O1 serotype Ogawa.Neither the pure cultures nor the original samples of both stains were positive with ctx gene.No v.cholerae was detected by colloidal gold immunochromatographic assay.Conclusions For V.cholerae detection in seafood samples,real time PCR can be first used for screen.Then conventional bacterial culture can be applied to isolate the strains in positive samples.In this way,the detection efficiency and detection rate can be improved.Meantime,the positive samples can be subjected to ctx gene detection.If it is positive as well,prevention and control measures should be taken to prevent cholera outbreaks.

关 键 词：霍乱弧菌 ctx基因 实时荧光PCR 常规细菌培养 胶体金法 

分 类 号：R378.3[医药卫生—病原生物学]