摘要
本文对ELISA间接法检测人轮状病毒(RV)血清抗体的方法学进行了探讨.结果表明,RV抗原能直接包被酶标反应板,与经免疫血清间接包被者无甚差异;RV抗原经EDTA、硫氰酸钾或盐酸胍预作用后再行包被,可提高抗原的包被效果.以ELISA间接法和间接双抗体夹心法同时检测34份血清标本,两者检出率无差异(P>0.05);进一步检测11份阳性血清的抗体效价,两法GMT分别为755.1和822.1,也无差异(P>0.05).进行ELISA间接法阻断试验,阻断率达95.6%;重复性试验表明变异系数在3.7～8.7%之间.因此,ELISA间接法敏感性高、特异性强和重复性好,可代替ELISA间接双抗体夹心法以检测RV血清抗体.
The methodology of indirect ELISA for the detection of rotavirus (RV) serum antibody has been investigated. The results indicate that the plates could be directly coated with RV antigen with no difference between the direct coating and indirect one by means of immune serum. The coating effect might be improved by pretreatment of RV particles with EDTA, KSCN or guanidine hydrochloride. No significant difference(P>0.05) existed between the detection rates by indirect ELISA and indirect sandwich when 34 serum samples were examined by both methods at the same time. Both methods were applied to determine the antibody titer of 11 positive sera. The GMT was 755.1 for indirect ELISA and 822.1 for indirect sandwich, without statistical difference (P>0.05). The blocking test of indirect ELISA was carried out, the blocking rate being 95.6%. The variance coefficient results measured by indirect ELISA in replicated samples ranged from 3.7% to 8.7%. Thus, indirect ELISA proved to possess high sensitivity, specificity and reproducibility, it can be used to replace indirect sandwich ELISA for the detection of serum antibody against RV.
