title: Immunostained plaque assay for detection and titration of rabies virus infectivity.
abstract: The fluorescent antibody test (FAT) is the most commonly used method for detection of the rabies virus (RV). The plaque assay can only be applied to fixed RVs, and cannot be used for street RVs. In this study, plaque formation allowing the determination of both fixed and street RVs was achieved using the immune plaque assay. The immune plaque assay carried out using both fixed and street RVs showed the formation of clear and countable plaques after immunostaining with anti-RV P monoclonal antibody and HRP-conjugated anti-mouse IgG. Plaque size increased with incubation time, and the plaque morphology differed according to viral strain. Fixed RVs had the dot-shaped regular plaque morphology and street RVs had the small irregular-shape plaque morphology. In addition, no significant differences were observed between the growth kinetics of the KGH strain when the virus was titrated using the FAT and the immune plaque assay. It allowed the successful detection and quantification of both street and fixed RVs through the production of clear, countable plaques, making it easy to obtain objective results. The assay is an applicable tool for the detection of RVs in various investigations, including virus neutralizing antibody testing, cell-to-cell spread, and viral drug sensitivity testing.
authors.affiliation: Division of Zoonoses, Korea National Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, Republic of Korea.
authors.forename: Jun-Sun
authors.initials: JS
authors.lastname: Park
authors.affiliation: Division of Zoonoses, Korea National Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, Republic of Korea.
authors.forename: Jihye
authors.initials: J
authors.lastname: Um
authors.affiliation: Department of Microbiology, College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju-si, Republic of Korea.
authors.forename: Young-Ki
authors.initials: YK
authors.lastname: Choi
authors.affiliation: Division of Zoonoses, Korea National Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, Republic of Korea.
authors.forename: Yeong Seon
authors.initials: YS
authors.lastname: Lee
authors.affiliation: Division of Zoonoses, Korea National Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, Republic of Korea.
authors.forename: Young Ran
authors.initials: YR
authors.lastname: Ju
authors.affiliation: Division of Zoonoses, Korea National Institute of Health, 187, Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do, Republic of Korea. Electronic address: tenksy@korea.kr.
authors.forename: Su Yeon
authors.initials: SY
authors.lastname: Kim
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D007118:Immunoassay / Q000379:methods; D051379:Mice; D008826:Microbial Sensitivity Tests; D009500:Neutralization Tests / Q000379:methods; D011818:Rabies / Q000175:diagnosis; D011820:Rabies virus / Q000254:growth & development* / Q000302:isolation & purification* / Q000472:pathogenicity; D010948:Viral Plaque Assay / Q000379:methods*
chemical_list: D000914:Antibodies, Viral
keywords: Detection; Immunostaining; Plaque assay; Rabies virus