施氏鲟病原嗜水气单胞菌ELISA快速检测方法的研究

以施氏鲟(Acipenser schrenchii)细菌性败血症病原菌嗜水气单胞菌为抗原,免疫兔获得高免血清,建立一种快速检测施氏鲟细菌性败血症病原菌嗜水气单胞菌的ELISA技术。结果显示:采用棋盘滴定法确定抗原和抗血清的最适工作浓度分别为107CFU/mL和1∶20000;病原菌检测灵敏度为每孔104CFU;抗血清与其它细菌标准菌株的交叉反应结果均呈阴性;阻断实验中的阻断率达66.88%;交叉反应和阻断实验的结果表明该方法具有较高的特异性。将该方法标准化后检测了22份人工感染后的施氏鲟和健康施氏鲟,阳性检测率分别为90.9%和13.6%,表明该技术不仅能够检测已发病的施氏鲟,而且能够检测带菌的施氏鲟。

Indirect ELISA method for detecting the hemorrhagic septicemia pathogen of sturgeon,Acipenser schrenckii

An indirect Enzyme-Linked Immunosorbent Assay (ELISA) for the rapid diagnosis of the hemorrhagic septicemia pathogen of sturgeon,Acipenser schrenckii Brandt,was developed.The best working concentration of antigen and antiserum determined by checkerboard titration was 10~7 CFU/mL and 1∶ 20000 respectively.The sensitivity of the serum was tested,and the lowest A.hydrophila suspension was 10~4 CFU.Cross reactions of antisera with other bacteria were detected,all results were negative.Inhibition rate was 66.88% in inhibition test.The cross assay and the inhibiton assay indicated that this method was differential.The method was optimized to detect the infected A.schrenchii and the normal ones,the positive detecting rate was 90.9% and 13.6% respectively.The results indicated that this assay can be used to detect not only the infected A.schrenchii,but also the carriers.This is very important to diagnose the disease in the early stage.