旋毛虫新生幼虫期特异性基因T668主要抗原表位短肽的筛选

为进一步从T668基因编码的蛋白中寻找到敏感性及特异性较高的抗原表位,对T668主要抗原表位区进行抗原表位作图,进一步设计了一套共16条覆盖整个抗原表位决定区的短肽,并与GST融合表达,用Western blot筛选具有高反应原性的短肽。结果显示,N5EM4、N5EM5、N5EM7、N5EM8、N5EM9、N5EM116条融合短肽与感染旋毛虫的猪血清有强阳性反应条带,但均表现出较明显的假阳性。将筛选出的6条短肽人工合成后与牛血清白蛋白偶联,利用Dot blot进一步验证,结果显示融合短肽N5EM11-BSA表现出良好的反应原性和特异性。此抗原短肽的发现对进一步确定T668蛋白虫体定位、功能及旋毛虫病诊断试剂盒的研发奠定了基础。

Screening of Major Antigenic Epitope Oligopeptide of Newborn Larva Stage-specific Gene T668 in Trichinella spiralis

In order to find highly sensitive and specific antigenic epitopes from protein which the T668 encoded,we made epitope mapping for the major epitope determin domain of T668 antigen,designed a set of 16 partial oligopeptide which covered the whole epitope determin domain and the GST-fusion expression,WB method was used for screening the highly antigenic oligopeptide.The results showed that 6 fusion peptides(N5EM4,N5EM5,N5EM7,N5EM8,N5EM9,N5EM11)had high postive reaction bands with the serum of swine infected with Trichinella spiralis,but showed obvious false-positive.So we synthesized the 6 antigenic oligopeptides and conjugated with BSA.Dot blot was used for verification.The results suggested that the oligopeptides-N5EM11-BSA showed better antigenicity and specificity.The discovery of antigenic oligopeptides lay the foundation for determining T668 protein polypide location,function and the study of diagnosis kit further.