Microbiology and Immunology Group, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
Clinical Research Development Unit, Yasuj University of Medical Sciences, Yasuj, Iran.
Viral Immunol. 2024 Aug;37(6):308-316. doi: 10.1089/vim.2024.0042. Epub 2024 Aug 2.
To develop polyomavirus VP1 recombinant protein-based immunoassay, the expression of two polyomavirus (Karolinska Institute Polyomavirus; KIPyV, and Washington University Polyomavirus; WUPyV) VP1s in insect cells was investigated using an improved baculovirus system (BacMagic). The reliability of the purified VP1 to serve as antigens in serological tests was confirmed by the establishment of an enzyme-linked immunosorbent assay (ELISA). Two panels of serum samples were used, with Panel I comprising 60 sera (20 KIPyV-positive, 20 WUPyV-positive, and 20 negative) and Panel II consisting of 134 sera with unknown status. The seroprevalence of KIPyV and WUPyV in the study population was determined to be 62% and 50%, respectively. Antibody-negative sera exhibited low reactivities in both ELISAs, whereas antibody-positive sera displayed high reactivity with median optical density values of 1.37 and 1.47 in the KIPyV and WUPyV ELISAs, respectively. The differences in seroreactivities between antibody positive and negative for each virus were statistically significant ( < 0.0001; with 95% confidence interval). The study suggests that seroconversion for KIPyV and WUPyV occurs in childhood, with KIPyV seropositivity reaching 70% and WUPyV seropositivity reaching 60% after the age of 5 years. Adult seroprevalence for polyomaviruses was high, with more than 64% and 51% of the adult population being seropositive for KIPyV and WUPyV, respectively. The constant prevalence of KIPyV and WUPyV antibody in the age groups suggested that this antibody persists for life. The fact that antibody titers were generally stable over time revealed a persistent infection of polyomaviruses in the human population. The insect cell-derived recombinant VP1-based ELISA has been demonstrated to be valuable as a serological assay, offering a valid, reliable, fast, nonlaborious, and economical procedure.
为了开发多瘤病毒 VP1 重组蛋白的免疫分析，本研究使用改良的杆状病毒系统（BacMagic），研究了两种多瘤病毒（卡罗林斯卡研究所多瘤病毒；KIPyV 和华盛顿大学多瘤病毒；WUPyV）VP1 在昆虫细胞中的表达情况。通过建立酶联免疫吸附试验（ELISA），证实了纯化的 VP1 作为血清学检测抗原的可靠性。使用了两批血清样本，Panel I 包括 60 份血清（20 份 KIPyV 阳性、20 份 WUPyV 阳性和 20 份阴性），Panel II 包括 134 份未知状态的血清。研究人群中 KIPyV 和 WUPyV 的血清流行率分别为 62%和 50%。抗体阴性血清在两种 ELISA 中均显示出低反应性，而抗体阳性血清在 KIPyV 和 WUPyV ELISA 中的平均光密度值分别为 1.37 和 1.47，显示出高反应性。两种病毒的抗体阳性和阴性血清之间的血清反应性差异具有统计学意义（<0.0001；95%置信区间）。研究表明，KIPyV 和 WUPyV 的血清转化发生在儿童时期，5 岁后 KIPyV 血清阳性率达到 70%，WUPyV 血清阳性率达到 60%。成人多瘤病毒的血清流行率较高，分别有超过 64%和 51%的成年人对 KIPyV 和 WUPyV 呈血清阳性。各年龄段 KIPyV 和 WUPyV 抗体的持续流行表明，这种抗体终生存在。抗体滴度随时间普遍稳定的事实表明，多瘤病毒在人群中持续感染。昆虫细胞衍生的重组 VP1 基于 ELISA 已被证明是一种有价值的血清学检测方法，提供了一种有效、可靠、快速、不费力和经济的检测方法。
