开发一种基于重组蛋白的免疫测定法，用于检测针对卡罗林斯卡研究所和华盛顿大学多瘤病毒的抗体。

IF 1.5 4区医学 Q4 IMMUNOLOGY

Viral immunology Pub Date : 2024-08-01 Epub Date: 2024-08-02 DOI:10.1089/vim.2024.0042

Bahman Abedi Kiasari, Mohammad Gholamnezhad, Amir Hossein Alipour, Fatemeh Hoda Fallah

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引用次数: 0

摘要

为了开发基于多瘤病毒 VP1 重组蛋白的免疫测定，研究人员使用改进的杆状病毒系统（BacMagic）在昆虫细胞中表达了两种多瘤病毒（卡罗林斯卡研究所多瘤病毒；KIPyV 和华盛顿大学多瘤病毒；WUPyV）的 VP1。通过建立酶联免疫吸附试验（ELISA），证实了纯化的 VP1 在血清学试验中作为抗原的可靠性。实验使用了两组血清样本，I 组包括 60 份血清（20 份 KIPyV 阳性、20 份 WUPyV 阳性和 20 份阴性），II 组包括 134 份状态不明的血清。经测定，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酶联免疫吸附试验中的反应度都很低，而抗体阳性血清的反应度很高，在 KIPyV 和 WUPyV 酶联免疫吸附试验中的光密度中值分别为 1.37 和 1.47。每种病毒的抗体阳性和阴性血清反应性差异均有统计学意义（P < 0.0001；置信区间为 95%）。研究表明，KIPyV 和 WUPyV 的血清转换发生在儿童时期，5 岁以后 KIPyV 血清阳性率达到 70%，WUPyV 血清阳性率达到 60%。成人的多瘤病毒血清阳性率很高，分别有超过 64% 和 51% 的成人对 KIPyV 和 WUPyV 呈血清阳性。KIPyV 和 WUPyV 抗体在各年龄组中的持续流行表明，这种抗体会终生存在。抗体滴度随着时间的推移基本保持稳定这一事实表明，多瘤病毒在人群中具有持续感染性。昆虫细胞衍生的基于重组 VP1 的 ELISA 被证明是一种有效、可靠、快速、不费力且经济的血清学检测方法。

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Development of a Recombinant Protein-Based Immunoassay for Detection of Antibodies Against Karolinska Institute and Washington University Polyomaviruses.

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 (p < 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.

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来源期刊

Viral immunology 医学-病毒学

CiteScore

3.60

自引率

0.00%

发文量

审稿时长

6-12 weeks

期刊介绍： Viral Immunology delivers cutting-edge peer-reviewed research on rare, emerging, and under-studied viruses, with special focus on analyzing mutual relationships between external viruses and internal immunity. Original research, reviews, and commentaries on relevant viruses are presented in clinical, translational, and basic science articles for researchers in multiple disciplines. Viral Immunology coverage includes: Human and animal viral immunology Research and development of viral vaccines, including field trials Immunological characterization of viral components Virus-based immunological diseases, including autoimmune syndromes Pathogenic mechanisms Viral diagnostics Tumor and cancer immunology with virus as the primary factor Viral immunology methods.