Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523, United States.
J Immunol Methods. 2013 Oct 31;396(1-2):74-86. doi: 10.1016/j.jim.2013.08.001. Epub 2013 Aug 14.
Microsphere immunoassays (MIAs) allow rapid and accurate evaluation of multiple analytes simultaneously within a biological sample. Here we describe the development and validation of domestic cat-specific MIAs for a) the quantification of total IgG and IgA levels in plasma, and b) the detection of IgG and IgA antibodies to feline immunodeficiency virus (FIV) capsid (CA) and surface (SU) proteins, and feline CD134 in plasma. These assays were used to examine the temporal antibody response of domestic cats infected with apathogenic and pathogenic FIVs, and domestic cats infected with parental and chimeric FIVs of varying pathogenicity. The results from these studies demonstrated that a) total IgG antibodies increase over time after infection; b) α-CA and α-SU IgG antibodies are detectable between 9 and 28 days post-infection and increase over time, and these antibodies combined represent a fraction (1.8 to 21.8%) of the total IgG increase due to infection; c) measurable α-CD134 IgG antibody levels vary among individuals and over time, and are not strongly correlated with viral load; d) circulating IgA antibodies, in general, do not increase during the early stage of infection; and e) total IgG, and α-CA and α-SU IgG antibody kinetics and levels vary with FIV viral strain/pathogenicity. The MIAs described here could be used to screen domestic cats for FIV infection, and to evaluate the FIV-specific or total antibody response elicited by various FIV strains/other diseases.
微球免疫分析(MIAs)允许在生物样本中同时快速准确地评估多种分析物。在这里,我们描述了用于 a)定量血浆中总 IgG 和 IgA 水平,以及 b)检测针对猫免疫缺陷病毒(FIV)衣壳(CA)和表面(SU)蛋白以及猫 CD134 的 IgG 和 IgA 抗体的国产猫特异性 MIA 的开发和验证。这些检测用于研究感染了致病性和非致病性 FIV 的家猫以及感染了不同致病性的亲本和嵌合 FIV 的家猫的时间抗体反应。这些研究的结果表明:a)总 IgG 抗体在感染后随时间增加;b)α-CA 和 α-SU IgG 抗体在感染后 9 至 28 天内可检测到,并随时间增加,这些抗体共同代表因感染而增加的总 IgG 的一部分(1.8 至 21.8%);c)个体之间和随时间变化的可测量的α-CD134 IgG 抗体水平不同,与病毒载量相关性不强;d)循环 IgA 抗体通常在感染早期不增加;e)总 IgG 以及 α-CA 和 α-SU IgG 抗体动力学和水平随 FIV 病毒株/致病性而变化。本文描述的 MIA 可用于筛选 FIV 感染的家猫,并评估各种 FIV 株/其他疾病引起的 FIV 特异性或总抗体反应。