摘要
以凡纳滨对虾红体病病原菌副溶血弧菌为抗原,免疫兔获得高免血清,建立一种快速检测凡纳滨对虾红体病病原菌副溶血弧菌的ELISA技术。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106CFU·mL-1和1∶2000;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75.88%;交叉反应和阻断实验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30份人工感染后的凡纳滨对虾和健康凡纳滨对虾,阳性检测率分别为93.3%和13.3%,表明该技术不仅能够检测已发病的凡纳滨对虾,而且能够检测带菌的凡纳滨对虾,这对于水产养殖业中疾病的早期诊断有着重要意义。
An indirect Enzyme-Linked Immtmosorbent Assay （ELISA） for the rapid diagnosis of Vibdo parahaemolyticus, the pathogen of the red body disease of Litopenaeus vannmei, has been developed. The best working concentration of antigen and antiserum were 10^6 CFU·mL^-1 and 1：2000 separately, determined by using checkerboard titration. The sensitivity of the serum was tested, and the lowest V. parahaemolyticus suspension was 10^4 CFU. Cross reactions of antisera with other bacteria were detected, all results were negative. Inhibition rate was 75.88 % in inhibiton test. The cross assay and the inhibiton assay indicate this method is differential. The method was optimized to detect the infected Litopenaeus vannamei and the normal ones, the positive detecting rate were 93.3 % and 13.3% separately. The results indicated that this assay can be used to detect not only the infected unhealthiness of L. vannamei, but also the carriers. This is very important to diagnose the disease in the early stage.
出处
《水产学报》
CAS
CSCD
北大核心
2006年第1期113-117,共5页
Journal of Fisheries of China
基金
国家863高技术研究发展计划项目(2002AA639470)
国家海洋局重点青年基金项目资助(2001303)
关键词
凡纳滨对虾
红体病
副溶血弧菌
间接ELISA技术
Litopenaeus vatmamei
red body disease
Vibrio parahaemolyticus
indirect enzyme-linked immunosorbent assay （ELISA）
