An enzyme-linked immunosorbent assay (ELISA) using a synthetic peptide for the E2 glycoprotein was developed for the serodiagnosis of Getah virus infection in horses. To identify an immunogenic epitope, a series of 20-mer peptides (n = 22) for the E2 protein was screened with pooled sera from horses infected with Getah virus. Peptide P11 (PTEEEIDMHTPPDIPDITLL) showed the strongest reaction. ELISA using P11 (E2-P11-ELISA) detected increased antibody levels in all seven experimentally infected horses and in five out of nine vaccinated horses. Out of 28 naturally infected horses, 25 were seronegative in their acute sera but turned seropositive in their convalescent sera. For the remaining three horses whose acute sera were seropositive, an endpoint method with serial dilutions detected a ≥ 4-fold increase in titer between paired sera. The concordance between E2-P11-ELISA and a virus-neutralization test in terms of seropositivity was assessed using a series of 220 horse sera, resulting in almost perfect agreement, with a kappa coefficient value of 0.865. E2-P11-ELISA had a sensitivity of 93.3% (95% CI 86.6-97.1%) and a specificity of 95.0% (95% CI 92.5-96.4%). This highly sensitive and specific E2-P11-ELISA should be useful for serodiagnosis of Getah virus infection in horses.

中文翻译：

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使用20聚体E2糖蛋白合成肽作为抗原，建立马中Getah病毒感染的酶联免疫吸附测定。

已开发出一种酶联免疫吸附测定法（ELISA），该合成物使用了E2糖蛋白的合成肽对马赫塔赫病毒感染进行血清学诊断。为了鉴定具有免疫原性的抗原决定簇，用来自感染了格塔病毒的马的合并血清筛选了一系列E2蛋白的20聚体肽（n = 22）。肽P11（PTEEEIDMHTPPDIPDITLL）显示出最强的反应。使用P11的ELISA（E2-P11-ELISA）在所有七只实验感染的马匹中和在九只接种疫苗的马匹中有五只检测到抗体水平的升高。在28头自然感染的马中，有25头急性血清呈血清阴性，而恢复期血清呈血清阳性。对于其余三匹急性血清呈阳性的马，采用连续稀释的终点法检测到成对血清之间的滴度增加≥4倍。使用一系列220马血清评估E2-P11-ELISA与病毒中和测试在血清阳性之间的一致性，得出几乎完美的一致性，κ系数值为0.865。E2-P11-ELISA的灵敏度为93.3％（95％CI 86.6-97.1％），特异性为95.0％（95％CI 92.5-96.4％）。这种高度灵敏，特异的E2-P11-ELISA对马格塔赫病毒感染的血清学诊断很有用。