应用胶体金免疫层析法检测水产品中霍乱弧菌的研究

Application Gold Immunchromatography Assay in Vibrio Cholerae Test of Aquatic Products

〔目的〕应用胶体金免疫层析法,研究水产品中霍乱弧菌的快速检测方法。〔方法〕通过模拟水产品棉拭子标本比对试验及现场应用比对试验,比较不同检验方法的差异。〔结果〕70份增菌液中霍乱弧菌理论浓度为1.5×10CFU/ml的模拟水产品棉拭子标本分别按检验方法1、检验方法2检验,同时检出阳性51份,检验方法2的检出率(87%)显著高于检验方法1的检出率(74%);30份增菌液中霍乱弧菌理论浓度为1.5×101CFU/ml的“模拟水产品棉拭子标本”按检验方法2进行检验的检测结果均为阳性。对2000份水产品进行检测的现场应用对照实验结果显示:检验方法3的检出率(0.4%)与检验方法1的检出率(0.3%)差异无显著性;胶体金试纸条实验的假阳性率为8%。〔结论〕在胶体金免疫层析法实验中,增菌液增菌前霍乱弧菌含量为1CFU/ml,通过增菌后可被检出;增菌液霍乱弧菌含量为10CFU/ml,增菌12h后均可被检出;水产品棉拭子标本增菌12h检测阴性者可报告阴性,阳性者则参照《霍乱防治手册》[1]进一步分离鉴定。鱼贝类霍乱弧菌污染菌数较少(103CFU/100g以下)[2],无法满足使用胶体金免疫层析法所要求的含菌量(106CFU/ml)[3],可以通过改良增菌培养液成分,以提高待检菌繁殖能力,在短时间内提高增菌液待检菌浓度,使胶体金免疫层析法的灵敏性高于或接近常规的分离鉴定法。

ObjectiveGICA was used to establish a new method of Rapid detection of Vibrio cholerae in aquatic product.MethodsCompare the difference in test with another base on compare test of the simulate aquatic product and the locale appliance.ResultsParting test for the 70 share of the aquatic product that it has vibrio cholerae for the chroma of exoterica is 1.510~0cfu per the millil(87%)iter in the increase bacterium(74%) by testl and test2,51 share both are positive by the testl and test2 at the same time,the checking rate of test2 is more higher than testl;They all are Positive for the 30 share of the aquatic product that it has vibrio cholerae for the chroma of exoterica is 1.510~1cfu per the milliliter in the increase bacterium by test2.The test result of the localeappliance for 2000 share of the aquatic product is show that:there were no significant difference between the positive percents of mehod3(0.4%)and method 1(0.3%);the false-positive percentage using GICA was 8%.Conclusionsbase on the GICA,It was checked that exoterica of vibrio cholerae is 1cfu per the milliliter in the increase bacterium;It all was checked that exoterica of vibrio cholerae is 10cfu per the milliliter in the increase bacterium after increase 12h;The negative may reported when It all was checked that aquatic product after increase 12h;The positive muse be baken more identify consultEnchiridion of Cholera prevention and cure^([1]).Vibrio cholerae in fish and shellfish are less,so that it cannot satisfy the request GICA^([3]),transit amend the bring up liquid,to improve growth of bacterium,to heighten chroma of bacterium,The sensitivity of GICA can be improved and overtop or be close to the conventional detection method of Vibrio cholerae.