title: Simple differentiation method of mumps Hoshino vaccine strain from wild strains by reverse transcription loop-mediated isothermal amplification (RT-LAMP).
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.
authors.affiliation: Kitasato Institute for Life Sciences, Laboratory of Viral Infection I, Shirokane 5-9-1, Minato-ku, Tokyo 108-8641, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Yoshida
authors.forename: Motoko
authors.initials: M
authors.lastname: Fujino
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Ota
authors.forename: Tsugunori
authors.initials: T
authors.lastname: Notomi
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nakayama
mesh_terms: D001483:Base Sequence; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry; D006801:Humans; D008582:Meningitis, Aseptic / Q000821:virology; D008969:Molecular Sequence Data; D009107:Mumps / Q000276:immunology / Q000821:virology; D009108:Mumps Vaccine / Q000235:genetics*; D009109:Mumps virus / Q000235:genetics*; D010309:Parotitis / Q000821:virology; D012150:Polymorphism, Restriction Fragment Length; D012367:RNA, Viral / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014613:Vaccines, Attenuated / Q000235:genetics
chemical_list: D009108:Mumps Vaccine; D012367:RNA, Viral; D014613:Vaccines, Attenuated; C047020:endodeoxyribonuclease ScaI; D015252:Deoxyribonucleases, Type II Site-Specific