摘要
以寄生于三疣梭子蟹血淋巴中的血卵涡鞭虫虫体为抗原,制备了多克隆血清抗体。抗体经健康梭子蟹血淋巴吸附处理后,间接ELISA检测效价达7 680。应用该抗体建立了血卵涡鞭虫的间接荧光抗体检测技术(IFAT)。采用常规显微镜检、PCR及IFAT 3种方法对采集的养殖青蟹、梭子蟹及海捕梭子蟹等18个样本进行了检测。检测结果显示:常规显微镜检阳性检出率为33.3%,而IFAT及PCR检测阳性率77.8%,符合率达100%;阳性虫体被染上黄绿色荧光,而正常梭子蟹血细胞则未被染色;可检测到血卵涡鞭虫不同生活阶段的营养体、腰鞭孢子及合孢体阶段。为血卵涡鞭虫的流行病学调查及生活史研究提供了简便实用的方法。
An indirect fluorescent antibody technique （IFAT） has been developed to detect Hematodinium sp. in the haemolymph of swimming crab and mud crab. The polyclonal antiserum was derived from rabbits immunized with trophonts of Hematodinium sp. in the diseased Portunus trituberculatus. Antiserum titer, detected by indirect ELISA, was 7 680 after reacted with the haemolymph of healthy swimming crab. 18 samples, including cultured swimming crab, mud crab and wild-caught swimming crab, were tested for Hematodinium sp. by microscopic observation, PCR and IFAT. And the detecting positive rate was 33.3 %, 77.8%, 77.8% respectively. The positive compatibility rate of the PCR and IFAT was 100%. Results showed that Hematodinium sp. was specific labelled with bright fluorescence under UV epifluorescence, while the haemocytes remained dark. Three different life stages, the trophont, dinospore, sporont of Hematodinium sp. , were also detected by IFAT. In conclusion, IFAT is rapid and specific for the detection of Hematodinium sp. , and it＇ s a practical method for the epidemiology investigation of marine crab and study on the life cycle of Hematodinium sp.
出处
《水产学报》
CAS
CSCD
北大核心
2009年第1期126-131,共6页
Journal of Fisheries of China
基金
浙江省科技厅院所专项(2006F4003)
浙江省自然科学基金项目(Y3080317,Y3080181)
浙江省科技厅人才培养项目(2006R20009)
关键词
血卵涡鞭虫
三疣梭子蟹
锯缘青蟹
间接荧光抗体
快速检测
Hematodiniurn sp.
Portunus trituberculatus
Scylla serrata
indirect fluorescent antibody technique
rapid detection
