Priest Jeffrey W, Moss Delynn M
Division of Foodborne, Waterborne, and Environmental Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
Methods Mol Biol. 2020;2052:61-85. doi: 10.1007/978-1-4939-9748-0_5.
For more than 35 years, various assay formats have been used to detect Cryptosporidium-specific antibodies in human and animal sera. Cryptosporidium parvum 17- and 27-kDa antigens, identified from invasive sporozoites, have been used in serologic antibody assays to identify individuals infected in outbreaks of diarrheal disease caused by this protozoan parasite and to monitor exposures in communities. During infection, immunoglobulin (Ig) A, IgM, and IgG responses are elicited by these immunodominant antigens, and the parasite-specific Ig responses diminish following the resolution of infection. Using the recombinant forms of the 17- and 27-kDa C. parvum antigens and the relatively recently developed multiplex bead assay (MBA), data from serologic antibody responses can be economically and efficiently acquired, especially when the Cryptosporidium assays are integrated with assays for antibody responses to antigens from other pathogens monitored in community-wide or nation-wide serosurveys. Here we describe the coupling of the C. parvum recombinant antigens to carboxylated polystyrene beads, the data acquisition and analysis of IgG antibodies bound to the coupled beads, and the quality control methods required for data validation using the Luminex/MBA system.
35多年来，人们使用了各种检测方法来检测人和动物血清中隐孢子虫特异性抗体。从小隐孢子虫侵袭性子孢子中鉴定出的17 kDa和27 kDa抗原已用于血清学抗体检测，以识别由这种原生动物寄生虫引起的腹泻病暴发中的感染者，并监测社区中的暴露情况。在感染过程中，这些免疫显性抗原会引发免疫球蛋白（Ig）A、IgM和IgG反应，感染消退后，寄生虫特异性Ig反应会减弱。使用17 kDa和27 kDa小隐孢子虫抗原的重组形式以及相对较新开发的多重微珠检测法（MBA），可以经济高效地获取血清学抗体反应数据，特别是当隐孢子虫检测与社区范围或全国范围血清学调查中监测的其他病原体抗原的抗体反应检测相结合时。在此，我们描述了小隐孢子虫重组抗原与羧化聚苯乙烯微珠的偶联、与偶联微珠结合的IgG抗体的数据采集和分析，以及使用Luminex/MBA系统进行数据验证所需的质量控制方法。
