Institut für Virologie, Zentrum für Infektionsmedizin, Robert von Ostertag-Haus, Freie Universität Berlin, Robert-von-Ostertag-Str. 7-13, 14163 Berlin, Germany.
J Virol Methods. 2013 Nov;193(2):667-73. doi: 10.1016/j.jviromet.2013.07.044. Epub 2013 Aug 5.
A peptide-based enzyme-linked immunosorbent assay (ELISA) for discrimination between serological responses to equine herpesvirus type 1 (EHV-1) and 4 (EHV-4) was developed. Three and four peptides for EHV-1 and EHV-4, respectively, were designed and studied initially in the ELISA using sera from foals infected experimentally. The most promising peptide pair, derived from EHV-1 glycoprotein E and EHV-4 glycoprotein G, was evaluated further using acute and convalescent sera from horses infected experimentally and naturally as well as a panel of horse field sera. Ten pre- and post-vaccination serum pairs were similarly tested in the type-specific ELISA. The peptide ELISA was able to identify horses which had been infected with EHV-1 or EHV-4 as derived from the results using acute and convalescent sera collected from natural outbreaks. When applied to a set of field samples, the assay proved robust with respect to determining the EHV-1 and EHV-4 antibody status. Also, the peptide ELISA was able to detect type-specific seroconversion for EHV-1 in vaccinated animals. With further validation, the EHV-1/EHV-4 peptide ELISA described in this study could serve as a reliable and cost-effective alternative to current methods for serological EHV-1 and EHV-4 diagnosis.
一种基于肽的酶联免疫吸附测定(ELISA)用于区分马疱疹病毒 1 型(EHV-1)和 4 型(EHV-4)的血清学反应已经被开发出来。分别针对 EHV-1 和 EHV-4 设计并研究了三个和四个肽,最初在使用实验感染的驹血清进行的 ELISA 中进行了研究。最有前途的肽对,源自 EHV-1 糖蛋白 E 和 EHV-4 糖蛋白 G,在使用实验和自然感染的马的急性和恢复期血清以及一组马野外血清进行了进一步评估。在针对特定类型的 ELISA 中,还对十对预接种和接种后血清对进行了类似的测试。该肽 ELISA 能够通过使用从自然爆发中收集的急性和恢复期血清的结果来识别已经感染了 EHV-1 或 EHV-4 的马。当应用于一组野外样本时,该测定在确定 EHV-1 和 EHV-4 抗体状态方面表现出很强的稳健性。此外,肽 ELISA 还能够检测接种动物的 EHV-1 型特异性血清转化。经过进一步验证,本研究中描述的 EHV-1/EHV-4 肽 ELISA 可以作为当前 EHV-1 和 EHV-4 血清学诊断方法的可靠且具有成本效益的替代方法。