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病毒样颗粒在血凝抑制试验中的新应用。

A Novel Application of Virus Like Particles in the Hemagglutination Inhibition Assay.

机构信息

Reference Laboratory for Veterinary Quality Control on Poultry Production (RLQP), Animal Health Research Institute (AHRI), Agriculture Research Center (ARC), Giza 12618, Egypt.

Medigen, Inc., Frederick, MD 21701, USA.

出版信息

Int J Mol Sci. 2024 Aug 11;25(16):8746. doi: 10.3390/ijms25168746.

DOI:10.3390/ijms25168746
PMID:39201433
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11354378/
Abstract

The hemagglutination inhibition (HI) assay is a traditional laboratory procedure for detection and quantitation of serum antibodies of hemagglutinating viruses containing the hemagglutinin (HA) gene. The current study aimed to investigate the novel use of virus like particles (VLP) as an antigen for the HI assay. VLPs were prepared from a strain of H5N1 using a baculovirus expression system. The VLPs were characterized using the hemagglutination test, Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting, and transmission electron microscopy. The comparative HI assay was performed using three different seed antigens: A/chicken/Mexico/232/94 (H5N2), A/chicken/Egypt/18-H/09(H5N1) and A/goose/Guangdong/1/1996(H5N1). The HI assay of serum antibody titrations using homologous antigens to these vaccinal seeds were compared to the VLP's antigens for the same serum. The HI titers were logically relevant to the similarity between VLP antigens and vaccinal seeds, indicating the VLPs behave similarly to the standard HI assay which uses inactivated whole virus as an antigen. VLPs could be considered as an alternative to the HI assay antigen as they show a relatedness between the similarity with vaccinal seed and serum antibodies. Compared to typical entire H5N1 viral antigen prepared in SPF eggs that require proper inactivation to avoid any public health risk, VLPs prepared in tissue culture, plants or insect cells are a safe, inexpensive and scalable alternative to inactivated whole virus antigen.

摘要

血凝抑制（HI）试验是一种传统的实验室程序，用于检测和定量含有血凝素（HA）基因的血凝病毒的血清抗体。本研究旨在探讨病毒样颗粒（VLP）作为 HI 试验抗原的新用途。使用杆状病毒表达系统从 H5N1 株制备 VLPs。使用血凝试验、十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）、Western blot 和透射电子显微镜对 VLPs 进行了表征。使用三种不同的种子抗原进行了比较 HI 试验：A/chicken/Mexico/232/94（H5N2）、A/chicken/Egypt/18-H/09（H5N1）和 A/goose/Guangdong/1/1996（H5N1）。使用同源抗原对这些疫苗种子进行血清抗体滴度的 HI 试验与相同血清的 VLP 抗原进行了比较。HI 滴度与 VLP 抗原和疫苗种子之间的相似性具有逻辑相关性，表明 VLPs 的行为类似于使用灭活全病毒作为抗原的标准 HI 试验。VLPs 可以被认为是 HI 试验抗原的替代品，因为它们在与疫苗种子和血清抗体的相似性之间表现出相关性。与在 SPF 鸡蛋中制备的典型整个 H5N1 病毒抗原相比，VLPs 制备于组织培养物、植物或昆虫细胞中是一种安全、廉价且可扩展的替代物，替代了灭活的全病毒抗原。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c7d/11354378/54b28df4a862/ijms-25-08746-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c7d/11354378/1ffb34010db7/ijms-25-08746-g001.jpg
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