Development Of Acolloidal Gold-based Immunochromatographic Lateral Flow Assay For The Detection Of Edwardsiella Tarda

Edwardsiella tardawas akind of important pathogenic bacteria in aquaculture and had a broad host range.Edwardsiellosis caused by E.tardahad a high incidence of mortality.It could infect many kinds of fish.Those infected fish showed symptoms of ascites,body surface haemorrhage,etc.It caused enormous losses in aquaculture.At present,the traditional detection methods includingobserving its morphologic,measuring its physiological and biochemical indexes,molecular biology method and ELISA. But these methods require expensive equipment and professional operation. Based on immune colloidal gold technique,colloidal gold-based immunochromatographic lateral flow assay was developed. Itwas a simple and rapid method for detecting E.tarda.The colloidal gold immunochromatographic strip forE.tardawas developed based on double antibodies sandwichELISA. In the strip, MAb 8H4-A3-B1 was labeled by 30 nm colloidal goldparticle as detection probe and rabbit PAb was as capture antibody. The strip had high specificity, without cross reaction with 与 Vibrio damsel ATCC33539、Vibrio parahaemolyticusKCTC2471、Vibrio vulnificusATCC2126、Vibrio alginnolyficus KCCM40513、Vibrio anguillarum KCTC2711、Vibrio Splendidus ATCC33125、Shewanella KCCM41821、Vibrio harveyi ACTT14126、Vibrio fluvialis KCCM40827. Its lower limit of detection was 1×106CFU/mL. The optimal detecting concentration was 1×108CFU/mL. The strip had good stability, it could still work over 6 month at 4℃ and only 4 month at RT.The colloidal gold immunochromatographic strip was a simple and rapid method for detecting E.tarda. The liquid sample was added at one side of the strip, the result was shown in 15 minutes.That was needless professional skills and special equipment. So it was very suitable for primary-level department of aquaculture.