Bleiholder Anne, Mühle Michael, Hechler Torsten, Bevins Sarah, vandeWoude Sue, Denner Joachim, Löchelt Martin
German Cancer Research Center, Focus Infection and Cancer, Heidelberg, Germany.
Vet Immunol Immunopathol. 2011 Oct 15;143(3-4):292-300. doi: 10.1016/j.vetimm.2011.06.007. Epub 2011 Jun 12.
The prevalence of feline foamy virus (FFV, spumaretrovirinae) in naturally infected domestic cats ranges between 30 and 80% FFV positive animals depending on age, sex and geographical region analyzed. Two serotypes have been reported for FFV designated FUV7-like and F17/951-like. Serotype-specific neutralization has been shown to correlate with sequence divergence in the surface (SU) domain of the envelope protein (Env). We analyzed a serum collection of 262 domestic cat sera from Germany using a GST-capture ELISA setup screening for Gag and Bet specific antibodies and identified 39% FFV positive animals. Due to the heterogeneity of the serological samples, cut-offs for Gag and Bet reactivity had to be experimentally determined since application of calculated cut-off values yielded some false-positive results; the new cut-off values turned out to be also fully applicable to a previous study. Using the already established FUV7 ElpSU antigen and the newly cloned and produced F17/951 ElpSU antigen, both consisting of the corresponding ectodomains of the envelope leader protein (Elp) and SU protein, we aimed at the detection of Env-specific antibodies and discrimination between the two known FFV serotypes within the diagnostic FFV ELISA. We validated the ElpSU antigens using cat reference sera of known serotype and screened with this assay domestic cat sera from Germany. Use of the FUV7- and F17/951 ElpSU antigens in ELISA resulted in the detection of Env-specific antibodies in both cat reference sera and sera from domestic cats in Germany, but failed to allow serotyping at the same time.
根据所分析的年龄、性别和地理区域,自然感染家猫的猫泡沫病毒(FFV,泡沫逆转录病毒亚科)的流行率在30%至80%的FFV阳性动物之间。FFV已报道有两种血清型,分别命名为FUV7样和F17/951样。血清型特异性中和作用已被证明与包膜蛋白(Env)表面(SU)结构域中的序列差异相关。我们使用谷胱甘肽-S-转移酶(GST)捕获酶联免疫吸附测定(ELISA)方法,对来自德国的一组262份家猫血清进行检测,筛查针对Gag和Bet的特异性抗体,结果确定39%的动物为FFV阳性。由于血清学样本的异质性,必须通过实验确定Gag和Bet反应性的临界值,因为应用计算得出的临界值会产生一些假阳性结果;新的临界值也被证明完全适用于之前的一项研究。我们使用已经确定的FUV7 ElpSU抗原和新克隆并制备的F17/951 ElpSU抗原,二者均由包膜前导蛋白(Elp)和SU蛋白的相应胞外结构域组成,旨在检测Env特异性抗体,并在诊断性FFV ELISA中区分两种已知的FFV血清型。我们使用已知血清型的猫参考血清对ElpSU抗原进行了验证,并使用该检测方法筛查了来自德国的家猫血清。在ELISA中使用FUV7和F17/951 ElpSU抗原,能够在猫参考血清和来自德国的家猫血清中检测到Env特异性抗体,但无法同时进行血清分型。