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来自合并感染和家猫野外分离株的血清样本在酶联免疫吸附测定中会非特异性结合猫免疫缺陷病毒及其他抗原。

Serum Samples from Co-Infected and Domestic Cat Field Isolates Nonspecifically Bind FIV and Other Antigens in Enzyme-Linked Immunosorbent Assays.

作者信息

Moskaluk Alex, Nehring Mary, VandeWoude Sue

机构信息

Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA.

出版信息

Pathogens. 2021 May 28;10(6):665. doi: 10.3390/pathogens10060665.

Abstract

We evaluated enzyme-linked immunosorbent assay (ELISA) specificity for measuring seroantibody responses to two types of retroviral infections in domestic cats: feline immunodeficiency virus (FIV) and feline foamy virus (FFV). We compared the seroreactivity of specific pathogen-free (SPF) cat sera, sera from SPF cats inoculated with either FIV or FFV, and field isolates (e.g., shelter or privately owned cats). Sera from SPF cats experimentally infected with the cognate virus had significantly lower background in both FIV and FFV ELISAs compared to sera from negative field isolates. ELISA values for SPF cats exposed to either FIV or FFV tended to have higher OD values on the opposite ELISA antigen plate. FIV nonspecific background absorbance was greater than that of FFV, and 10 of 15 sera samples from FIV seronegative field samples were measured in the indeterminant range. These findings highlight that exposure to off-target pathogens elicit antibodies that may nonspecifically bind to antigens used in binding assays; therefore, validation using sera from SPF animals exposed during controlled infection results in the setting of a cutoff value that may be inappropriately low when applied to field samples. Our work also suggests that infection of domestic cats with pathogens other than FIV results in antibodies that cross-react with the FIV Gag antigen.

摘要

我们评估了酶联免疫吸附测定(ELISA)在检测家猫对两种逆转录病毒感染的血清抗体反应方面的特异性:猫免疫缺陷病毒(FIV)和猫泡沫病毒(FFV)。我们比较了无特定病原体(SPF)猫血清、接种FIV或FFV的SPF猫血清以及野外分离株(如收容所或私人饲养的猫)的血清反应性。与来自阴性野外分离株的血清相比,经同源病毒实验感染的SPF猫血清在FIV和FFV ELISA中背景均显著较低。暴露于FIV或FFV的SPF猫在相反的ELISA抗原板上的ELISA值往往具有更高的光密度(OD)值。FIV的非特异性背景吸光度大于FFV,15份FIV血清阴性野外样本中的10份血清样本在不确定范围内被检测到。这些发现突出表明,暴露于非目标病原体可引发抗体,这些抗体可能与结合测定中使用的抗原发生非特异性结合;因此,使用在受控感染期间暴露的SPF动物的血清进行验证会导致设定一个临界值,当应用于野外样本时该临界值可能过低。我们的工作还表明,家猫感染FIV以外的病原体可导致与FIV Gag抗原发生交叉反应的抗体产生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb2a/8226436/44c63862b4da/pathogens-10-00665-g001.jpg

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