Alcaraz C, De Diego M, Pastor M J, Escribano J M
Instituto Nacional de Investigaciones Agrarias, Departamento de Sanidad Animal, Madrid, Spain.
J Vet Diagn Invest. 1990 Jul;2(3):191-6. doi: 10.1177/104063879000200307.
A radioimmunoprecipitation assay (RIPA) has been developed for detection of antibody to African swine fever virus (ASFV) and compared with the immunoblot assay with regard to sensitivity and specificity. Two hundred seven field sera, obtained from pigs in Spain from different geographic areas between 1975 and 1986, that were positive by ASFV enzyme-linked immunosorbent assay (ELISA) were also analysed by immunoblot assay and RIPA. By serum dilution experiments, the RIPA appeared at least as sensitive as the ELISA and immunoblotting tests, although ELISA and RIPA detected antibodies to ASFV earlier in natural infection than did the immunoblot assay, as disclosed by animal inoculation studies. The most antigenic ASFV-induced proteins in natural infection detected by RIPA were the viral proteins p243, p172, p73, p25.5, p15, and p12 and the infection proteins p30 and p23.5. In the immunoblot assay, the proteins that were most reactive with the same sera were the viral protein p25.5 and the infection proteins p30, p25, and p21.5. Only 1 serum, from an animal infected with ASFV, was negative by immunoblot assay but showed a positive result by RIPA. A modification of conventional RIPA was performed using a dot transference of immunoprecipitated proteins to a nitrocellulose filter. This modification simplified the conventional RIPA procedures by eliminating the electrophoresis of immunoprecipitated proteins without affecting sensitivity and specificity. The ease of use, specificity, and the sensitivity comparable to that of the immunoblot assay make the RIPA a useful confirmatory assay for sera that yield conflicting results in other ASFV antibody assays.
已开发出一种放射免疫沉淀试验（RIPA）用于检测非洲猪瘟病毒（ASFV）抗体，并在敏感性和特异性方面与免疫印迹试验进行了比较。对1975年至1986年间从西班牙不同地理区域的猪身上采集的207份经ASFV酶联免疫吸附试验（ELISA）检测呈阳性的现场血清，也进行了免疫印迹试验和RIPA分析。通过血清稀释实验，RIPA的敏感性至少与ELISA和免疫印迹试验相当，不过动物接种研究表明，在自然感染中，ELISA和RIPA比免疫印迹试验更早检测到ASFV抗体。RIPA在自然感染中检测到的最具抗原性的ASFV诱导蛋白是病毒蛋白p243、p172、p73、p25.5、p15和p12以及感染蛋白p30和p23.5。在免疫印迹试验中，与相同血清反应最强的蛋白是病毒蛋白p25.5以及感染蛋白p30、p25和p21.5。仅1份来自感染ASFV动物的血清，免疫印迹试验结果为阴性，但RIPA结果呈阳性。采用将免疫沉淀蛋白点转移至硝酸纤维素滤膜的方法对传统RIPA进行了改良。这种改良通过省去免疫沉淀蛋白的电泳简化了传统RIPA程序，且不影响敏感性和特异性。RIPA使用方便、特异性强且敏感性与免疫印迹试验相当，使其成为对在其他ASFV抗体检测中结果相互矛盾的血清进行确证检测的有用方法。
