猴B病毒BVgD-多肽ELISA检测方法的建立

Developing of The BVgD-Peptide ELISA for Detection of Antibodies Against Monkey B Virus

目的建立猴B病毒抗体BVgD-多肽ELISA检测方法。方法以Western-blot分析猴B病毒糖蛋白D(BVgD)上的多肽抗原决定簇(BVgD-多肽)的抗原性,采用方阵滴定法,确定ELISA法的最佳实验条件。用灭活BV全病毒、HSV-1和BVgD-多肽三种抗原系统对猴血清样品进行平行检测,比较分析了三种系统的抗原性特点。并且将检测结果与美国BV抗体检测试剂盒的检测结果进行了比较。结果建立了猴B病毒抗体的BVgD-多肽ELISA法,研究结果表明本方法与BV全病毒和以HSV-1病毒为抗原的试剂盒相比,敏感性较低、特异性较好,避免了制备BV抗原对实验室要求高和HSV-1抗原引起的非特异性问题。检测结果与美国实验室BV抗体检测结果的符合率达85%。

Objective To develop ELISA for detection of antibodies against monkey B virus using BVgD-Peptide as antigen. Methods Antigenicity of BVgD-Peptide antigenic determinant was analyzed by the Western-blot method. The EHSA method was established by optimization of reaction condition and reagent selection. The parallel detection of monkey sera was performed by using three different ELISA kits using complete BV, HSV-1 and BVgD-Peptide as antigens. The detection results obtained by ELISA kit developed in the study were compared with the results obtained by ELISA kit made in USA. Results Compared with ELISA using as complete BV and HSV-1 antigens, ELISA using BVgD-Peptide as antigen was less sensitive but more specific. 85% of the detection results obtained by ELISA kit developed in the study were coincident with those obtained form Laboratory of America. Conclusion ELISA for detection of antibodies against monkey B virus using BVgD-Peptide as antigen has been developed, and this ELISA is more specific than ELSA using complete BV and HSV-1 as antigens.