Abstract
Mumps viruses show diverse cytopathic effects (CPEs) of infected cells and viral plaque formation (no CPE or no plaque formation in some cases) depending on the viral strain, highlighting the difficulty in mumps laboratory studies. In our previous study, a new sialidase substrate, 2-(benzothiazol-2-yl)-4-bromophenyl 5-acetamido-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosidonic acid (BTP3-Neu5Ac), was developed for visualization of sialidase activity. BTP3-Neu5Ac can easily and rapidly perform histochemical fluorescent visualization of influenza viruses and virus-infected cells without an antiviral antibody and cell fixation. In the present study, the potential utility of BTP3-Neu5Ac for rapid detection of mumps virus was demonstrated. BTP3-Neu5Ac could visualize dot-blotted mumps virus, virus-infected cells, and plaques (plaques should be called focuses due to staining of infected cells in this study), even if a CPE was not observed. Furthermore, virus cultivation was possible by direct pick-up from a fluorescent focus. In conventional methods, visible appearance of the CPE and focuses often requires more than 6 days after infection, but the new method with BTP3-Neu5Ac clearly visualized infected cells after 2 days and focuses after 4 days. The BTP3-Neu5Ac assay is a precise, easy, and rapid assay for confirmation and titration of mumps virus.
Highlights
Acute infection with mumps virus is manifested by demonstrable swelling of the parotid glands and several complications including aseptic meningitis, encephalitis, deafness, orchitis, oophoritis, and pancreatitis
We examined whether mumps virus could be cultivated by direct pick-up from fluorescent focuses stained with BTP3-Neu5Ac
To confirm that BTP3-Neu5Ac can visualize sialidase activity of mumps virus, dot-blotted mumps virus on a polyvinyl difluoride (PVDF) membrane was incubated with 10 μM BTP3-Neu5Ac at 37°C for 15 min
Summary
Acute infection with mumps virus is manifested by demonstrable swelling of the parotid glands and several complications including aseptic meningitis, encephalitis, deafness, orchitis, oophoritis, and pancreatitis. Clinical diagnosis is based on parotid swelling [1,2,3]. Laboratory detection and diagnosis are based on isolation of the virus, detection of the viral gene, or serological confirmation (generally the presence of anti-mumps virus IgM antibodies). Easy and Rapid Assay for Mumps Virus Detection.
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