Use of the particle agglutination/particle agglutination inhibition test for antigenic analysis of SARS‐CoV‐2
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Published:2023-02
Issue:2
Volume:17
Page:
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ISSN:1750-2640
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Container-title:Influenza and Other Respiratory Viruses
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language:en
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Short-container-title:Influenza Resp Viruses
Author:
Kobayashi Jun1,
Matsuyama Shutoku1,
Shirakura Masayuki2,
Arita Tomoko3,
Suzuki Yasushi3,
Asanuma Hideki3,
Watanabe Shinji2,
Hasegawa Hideki123,
Nakamura Kazuya2ORCID
Affiliation:
1. Laboratory of Acute Respiratory Virus, Research Center for Influenza and Respiratory Viruses National Institute of Infectious Diseases Tokyo Japan
2. Laboratory of Influenza Virus Surveillance, Research Center for Influenza and Respiratory Viruses National Institute of Infectious Diseases Tokyo Japan
3. Laboratory of Vaccine Seed Virus Development, Research Center for Influenza and Respiratory Viruses National Institute of Infectious Diseases Tokyo Japan
Abstract
AbstractBackgroundThe antigenicity of SARS‐CoV‐2 is a critical issue for the effectiveness of the vaccine, and thus, it should be phenotypically evaluated by serological assays as new field isolates emerge. The hemagglutination/hemagglutination inhibition (HA/HI) tests are well known as a representative method for antigenic analysis of influenza viruses, but SARS‐CoV‐2 does not agglutinate human or guinea pig red blood cells. Therefore, the antigenic analysis requires complicated cell‐based assays using special equipment such as plate reader or ELISPOT analyzer.MethodsBased on the HA/HI tests for influenza viruses, we developed the particle agglutination/particle agglutination inhibition (PA/PAI) test to easily and rapidly quantify the virus and antibody using human angiotensin‐converting enzyme 2 (hACE2)‐bound latex beads. The virus titers were determined by mixing the beads and the virus from culture supernatant, settling it overnight, and then observing the sedimentation/agglutination pattern (PA test). The neutralization antibody titers were determined by mixing virus‐infected hamster antisera in addition to the beads and virus (PAI test).ResultsThe PA titer was positively correlated with the plaque‐forming units. The PAI titer using the hamster antisera clearly revealed the antigenic difference between the omicron and previous variants. The antigenic differences were supported by the results shown in other methods.ConclusionsThe PAI test is an easy and rapid method to analyze the antigenicity of SARS‐CoV‐2.
Funder
Japan Agency for Medical Research and Development
Uehara Memorial Foundation
Subject
Infectious Diseases,Public Health, Environmental and Occupational Health,Pulmonary and Respiratory Medicine,Epidemiology