Centro de Virología Animal, Instituto de Ciencia y Tecnología Dr. Cesar Milstein, CONICET, Saladillo 2468, C1440FFX, Ciudad de Buenos Aires, Argentina.
J Virol Methods. 2010 Jun;166(1-2):21-7. doi: 10.1016/j.jviromet.2010.02.011. Epub 2010 Feb 17.
A single serum-dilution liquid phase ELISA (slpELISA) was standardized to be used for serological evaluation of herd immunity against foot-and-mouth disease. The absorbance value at a dilution 1:64 of each serum sample was interpolated in a standard curve by plotting the antibody titers of six control sera determined by end point dilution liquid phase ELISA (lpELISA), against the absorbance values for the same control sera at 1:64 dilutions. A straight line was obtained by linear regression analysis (r>0.90) in the titer range of 1.40-2.40. The reliability of the antibody titers was confirmed by the simultaneous titration of 60 cattle sera by slpELISA and lpELISA, which showed an acceptable correlation (R(2)>0.87) for viral strains A24/Cruzeiro, A/Argentina/01, O1/Campos and C3/Indaial. Titers obtained by both methods were not significantly different (p>0.05), thus confirming that slpELISA could be used successfully to replace the conventional serial dilution ELISA for the assessment of protection status of cattle in epidemiological studies. In addition, this quantitative slpELISA provides an adequate method for monitoring the effectiveness of vaccination campaigns and is also suitable for the assessment of seroconversion of naive animals during early stages of infection.
建立了一种血清倍比稀释液相酶联免疫吸附试验(slpELISA),用于评估口蹄疫群体免疫的血清学效果。将每个血清样本在 1:64 稀释度下的吸光度值通过线性回归分析(r>0.90),绘制出 6 份对照血清的终点稀释液相酶联免疫吸附试验(lpELISA)抗体效价与同一份对照血清在 1:64 稀释度下的吸光度值之间的标准曲线,将每份血清在 1:64 稀释度下的吸光度值代入该标准曲线,即可得到效价。用 slpELISA 和 lpELISA 同时对 60 份牛血清进行滴定,确认了抗体效价的可靠性,结果表明 A24/Cruzeiro、A/Argentina/01、O1/Campos 和 C3/Indaial 病毒株的相关性良好(R(2)>0.87)。两种方法的效价差异无统计学意义(p>0.05),这表明 slpELISA 可成功替代常规的系列稀释 ELISA 用于评估流行病学研究中牛的保护状态。此外,这种定量 slpELISA 为监测疫苗接种效果提供了一种充分的方法,也适用于评估感染早期未免疫动物的血清转化情况。