Gupta G D, Lakritz J, Saville W J, Livingston R S, Dubey J P, Middleton J R, Marsh A E
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri, Columbia, Missouri 65211, USA.
J Parasitol. 2004 Oct;90(5):1027-33. doi: 10.1645/0022-3395(2004)090[1027:AEOARB]2.0.CO;2.
Sarcocystis neurona is the primary parasite associated with equine protozoal myeloencephalitis (EPM). This is a commonly diagnosed neurological disorder in the Americas that infects the central nervous system of horses. Current serologic assays utilize culture-derived parasites as antigen. This method requires large numbers of parasites to be grown in culture, which is labor intensive and time consuming. Also, a culture-derived whole-parasite preparation contains conserved antigens that could cross-react with antibodies against other Sarcocystis species and members of Sarcocystidae such as Neospora spp., Hammondia spp., and Toxoplasma gondii. Therefore, there is a need to develop an improved method for the detection of S. neurona-specific antibodies. The sera of infected horses react strongly to surface antigen 1 (SnSAG1), an approximately 29-kDa protein, in immunoblot analysis, suggesting that it is an immunodominant antigen. The SnSAG1 gene of S. neurona was cloned, and recombinant S. neurona SAG1 protein (rSnSAG1-Bac) was expressed with the use of a baculovirus system. By immunoblot analysis, the rSnSAG1-Bac antigen detected antibodies to S. neurona from naturally infected and experimentally inoculated equids, cats, rabbit, mice, and skunk. This is the first report of a baculovirus-expressed recombinant S. neurona antigen being used to detect anti-S. neurona antibodies in a variety of host species.
肉孢子虫是与马属动物原虫性脑脊髓炎(EPM)相关的主要寄生虫。这是一种在美洲常见的被诊断出的神经系统疾病,会感染马的中枢神经系统。目前的血清学检测使用培养获得的寄生虫作为抗原。这种方法需要在培养物中培养大量寄生虫,既耗费人力又耗时。此外,培养获得的全寄生虫制剂包含保守抗原,可能会与针对其他肉孢子虫物种以及肉孢子虫科成员(如新孢子虫属、哈蒙德孢子虫属和弓形虫)的抗体发生交叉反应。因此,需要开发一种改进的方法来检测肉孢子虫特异性抗体。在免疫印迹分析中,感染马的血清对表面抗原1(SnSAG1,一种约29 kDa的蛋白质)反应强烈,这表明它是一种免疫显性抗原。克隆了肉孢子虫的SnSAG1基因,并使用杆状病毒系统表达了重组肉孢子虫SAG1蛋白(rSnSAG1-Bac)。通过免疫印迹分析,rSnSAG1-Bac抗原检测到了来自自然感染和实验接种的马、猫、兔、小鼠和臭鼬体内针对肉孢子虫的抗体。这是首次报道杆状病毒表达的重组肉孢子虫抗原用于检测多种宿主物种中抗肉孢子虫抗体。