SPA胶体金免疫层析方法检测PCV2抗体的初步建立及应用

Establishment and Application of SPA Colloidal Gold Immuno-chromatography for Detection of Antibodies against PCV-2

采用柠檬酸三钠还原法制备胶体金颗粒,并将其标记SPA,将金标SPA包被至玻璃纤维膜上制备金标垫,将纯化的Cap蛋白、兔抗SPA抗体分别包被至硝酸纤维素膜的检测线与质控线上,建立了一种检测猪圆环病毒2型抗体的胶体金免疫层析方法。检测结果显示,组装的检测卡只与圆环病毒阳性血清发生特异性反应,检测的灵敏度可以稀释到100倍。用此检测卡与商品化ELISA试剂盒对86份临床血清进行平行检测,两者的检测阳性率分别为65.12%和70.93%。该方法检测速度快,15min内可出结果,特异性强,操作简便,可广泛应用于基层。

Staphylococcal protein A（SPA）labeled with colloidal gold particles prepared by trisodium citrate reduction and the labeled SPA was coated onto glass fibrous membrane to prepare gold labeled pad. The purified Cap protein of porcine circovirus 2 and rabbit anti-SPA antibody were coated respectively to the test line and quality control line on nitrate cellulose membrane,so as to establish a colloidal gold immuno-chromatography for detection of antibodies against porcine circovirus-2. The results indicated that the assembled test card reacted exclusively with PCV-2 positive sera and the test sensitivity was up-to 100 fold dilutions. 86 clinical samples were tested both with the test card and the commercial ELISAkid,resulting in positive rates of 65.12% and 70.93% respectively. The method was rapid,specific and simple in operation,and could be extended at grass-roots.