Qureshi T, Wagner G G, Drawe D L, Davis D S, Craig T M
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station 77843, USA.
Vet Parasitol. 1995 Jul;58(4):357-63. doi: 10.1016/0304-4017(94)00731-q.
Fasciola hepatica is a parasite of cattle (Bos taurus), but not of white-tailed deer (Odocoileus virginianus), while Fascioloides magna is a parasite of white-tailed deer which also infects cattle as dead-end host. Adult parasites were collected from naturally infected white-tailed deer or cattle. Excretory-secretory proteins (ESP) were obtained from each parasite. Protein banding patterns were analysed on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and probed using sera from experimentally infected deer of cattle using enzyme-linked immunoelectrotransfer blot (EITB, also known as Western blot) analysis. Protein banding patterns of the two species were different. EITB analysis of Fascioloides magna ESP using sera from Fascioloides magna infected deer or cattle identified three bands of approximately 17, 22 and 27 kDa of which the 27 kDa antigen cross-reacted with sera from Fasciola hepatica infected cattle. EITB analysis of Fasciola hepatica ESP probed with sera from Fasciola hepatica infected cattle identified three bands of approximately 15, 26 and 46 kDa. The 46 and 26 kDa ESP cross-reacted with sera from Fascioloides magna infected cattle, but not with sera from Fascioloides magna infected deer. The band at 15 kDa which reacted specifically for Fasciola hepatica infected cattle sera consisted of two protein bands close to each other as seen on the SDS-PAGE gel. The EITB reaction at approximately 17 kDa and 22 kDa of Fascioloides magna ESP, and at approximately 15 kDa of Fasciola hepatica ESP can be used for species specific diagnosis.
肝片吸虫是牛（Bos taurus）的寄生虫，但不是白尾鹿（Odocoileus virginianus）的寄生虫，而巨片吸虫是白尾鹿的寄生虫，它也会感染牛，但牛作为终末宿主。从自然感染的白尾鹿或牛中收集成年寄生虫。从每种寄生虫中获取排泄分泌蛋白（ESP）。在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）上分析蛋白质条带模式，并使用酶联免疫电转移印迹（EITB，也称为蛋白质印迹）分析，用实验感染的牛或鹿的血清进行检测。这两个物种的蛋白质条带模式不同。用感染巨片吸虫的鹿或牛的血清对巨片吸虫ESP进行EITB分析，鉴定出三条约17、22和27 kDa的条带，其中27 kDa抗原与感染肝片吸虫的牛的血清发生交叉反应。用感染肝片吸虫的牛的血清对肝片吸虫ESP进行EITB分析，鉴定出三条约15、26和46 kDa的条带。46 kDa和26 kDa的ESP与感染巨片吸虫的牛的血清发生交叉反应，但不与感染巨片吸虫的鹿的血清发生交叉反应。在SDS - PAGE凝胶上可见，15 kDa处与感染肝片吸虫的牛血清特异性反应的条带由两条彼此靠近的蛋白质条带组成。巨片吸虫ESP约17 kDa和22 kDa处以及肝片吸虫ESP约15 kDa处的EITB反应可用于物种特异性诊断。
