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J Immunol Methods. 2017 Jul;446:15-20. doi: 10.1016/j.jim.2017.03.020. Epub 2017 Apr 1.

A rapid, simple, and accurate plaque assay for human respiratory syncytial virus (HRSV).

Author information

1
Department of Biomedical Engineering, College of Medicine, Kyung Hee University, Seoul, Republic of Korea.
2
Department of Biomedical Science, Graduate School, Kyung Hee University, Seoul, Republic of Korea.
3
Department of Emergency, Yanbian University Hospital, Yanji City, China.
4
Department of Pathology, Kyung Hee University Medical Center, Seoul, Republic of Korea.
5
Department of Medical Zoology, Kyung Hee University School of Medicine, Seoul, Republic of Korea. Electronic address: fsquan@khu.ac.kr.

Abstract

Plaque assays of human respiratory syncytial virus (HRSV) are time-consuming, requiring 4 to 7 days for plaque formation and several hours for dye staining. Here, we describe a simple method by which RSV plaques can be visualized and counted with the naked eye only 2 days after infection of HEp-2 cells. In this assay, the infected cells are stained with monoclonal antibodies and the plaques are developed using diaminobenzidine (DAB). We tested the accuracy of this new plaque assay by comparing the results obtained on days 1, 2, 3, 4, 5, and 6 post-infection. The whole procedure is significantly simpler than the traditional method, with an immunostaining process of around 1.5h. Our method is rapid, accurate, and simple; thus, it has the potential to significantly contribute to studies related to RSV disease.

KEYWORDS:

Human respiratory syncytial virus; Immunostaining; Plaque assay

PMID:
28373028
DOI:
10.1016/j.jim.2017.03.020
[Indexed for MEDLINE]

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