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Simple differentiation method of mumps Hoshino vaccine strain from wild strains by reverse transcription
Naoko Yoshida1, Motoko Fujino, Yoshinori Ota
1Kitasato Institute for Life Sciences, Laboratory of Viral Infection I, Shirokane 5-9-1, Minato-ku, Tokyo 108-8641, Japan.
Abstract:
Mumps virus is still circulating and annual mumps outbreaks occur with fluctuating magnitudes in Japan. Aseptic meningitis has been reported after vaccination and it would be of importance to determine whether this was related to the vaccination. The objective of this study was to develop a sensitive, specific and rapid diagnostic method for the differentiation of the Hoshino vaccine strain from circulating wild types. We developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method of the hemagglutinin neuraminidase (HN) region for the detection of mumps virus genome from clinical samples. The typical ladder pattern disappeared after the LAMP products of the Hoshino vaccine strain were digested with ScaI, but those of wild types were not cut by ScaI. We obtained 19 cerebro spinal fluids (CSF) from the patients with aseptic meningitis and 17 salivary swab samples from the patients with acute parotitis after mumps vaccination, in which one case was complicated with orchitis. Mumps virus genome was detected in 18 CSF samples and in all NPS by RT-LAMP. The Hoshino vaccine strain was identified in 16 out of 18 CSF RT-LAMP positives and in 11 out of 17 NPS samples and the remaining samples were identified as wild types. RT-LAMP followed by ScaI digestion is a sensitive, simple and rapid differential method and useful for laboratory surveillance for vaccine-adverse events.
Insights
A new diagnostic method can distinguish the Hoshino mumps vaccine strain from wild mumps virus in clinical samples. This rapid test aids in investigating potential vaccine-adverse events like aseptic meningitis.
Area of Science:
- Virology
- Immunology
- Molecular Diagnostics
Background:
- Mumps virus circulates globally, causing annual outbreaks in Japan.
- Aseptic meningitis is a reported adverse event following mumps vaccination, necessitating differentiation between vaccine and wild strains.
Purpose of the Study:
- To develop a sensitive, specific, and rapid diagnostic method to differentiate the Hoshino mumps vaccine strain from wild-type strains.
- To assess the utility of this method in clinical samples from patients experiencing vaccine-adverse events.
Main Methods:
- Developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) targeting the hemagglutinin neuraminidase (HN) region of mumps virus.
- Utilized ScaI restriction enzyme digestion to differentiate between Hoshino vaccine strain and wild-type strains based on LAMP product patterns.
- Analyzed cerebrospinal fluid (CSF) and salivary swab samples from patients with aseptic meningitis and acute parotitis post-vaccination.
Main Results:
- Mumps virus genome was detected in 18/19 CSF samples and 17/17 NPS samples using RT-LAMP.
- The Hoshino vaccine strain was identified in 16/18 CSF and 11/17 NPS samples.
- Wild-type mumps strains were identified in the remaining positive samples, indicating co-circulation or vaccine strain identification.
Conclusions:
- RT-LAMP followed by ScaI digestion is a sensitive, simple, and rapid differential diagnostic method for mumps virus.
- This method is valuable for laboratory surveillance of vaccine-adverse events and understanding mumps epidemiology.
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