title: Development of an efficient method for recovery of Puumala and Puumala-related viruses by inoculation of Mongolian gerbils.
abstract: Puumala (PUU) virus and PUU-related viruses are difficult to isolate in cell culture. To determine whether animal inoculation would be a better alternative for virus recovery, the Sotkamo strain of PUU virus was inoculated into several animal species. Newborn Mongolian gerbils (MGs), mice, and rats were infected with the Sotkamo strain by intracerebral (ic), intraperitoneal (ip), and subcutaneous (sc) inoculation. Antibodies to PUU appeared in MGs at 30 days post-infection (dpi), and in mice and rats at 15 dpi. Interestingly, virus appeared at 7 dpi in lung and brain of MGs inoculated via ic and ip routes. Virus was detected in all tested tissues of MGs at 15 dpi, with a peak level of 1.36 x 10 (5) focus forming units (FFU)/g in brain tissue. The virus titer declined with the onset of the antibody response and became undetectable by 75 dpi, when the antibody titer reached the maximum level. The appearance of the virus in mice and rats was delayed as compared to MGs, and the virus titer was apparently lower, at approximately 4 to 8 x 10(3) FFU/g, at 15 dpi. In addition, lung homogenates of antibody-positive Clethrionomys (C.) rufocanus (captured in Tobetsu, Hokkaido, Japan) were inoculated into MGs by the ic route. PUU-related viral RNA was detected at 16 dpi in the brains of MG inoculated with the lung homogenate, and antibodies were detected at 45 dpi. These findings indicate that newborn MG inoculation is an efficient method to recover PUU and PUU-related viruses.
authors.affiliation: Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
authors.forename: Nandadeva
authors.initials: N
authors.lastname: Lokugamage
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kariwa
authors.forename: Kumari
authors.initials: K
authors.lastname: Lokugamage
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Hagiya
authors.forename: Hironobu
authors.initials: H
authors.lastname: Miyamoto
authors.forename: Masahiro A
authors.initials: MA
authors.lastname: Iwasa
authors.forename: Koichi
authors.initials: K
authors.lastname: Araki
authors.forename: Kumiko
authors.initials: K
authors.lastname: Yoshimatsu
authors.forename: Jiro
authors.initials: J
authors.lastname: Arikawa
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Mizutani
authors.forename: Ikuo
authors.initials: I
authors.lastname: Takashima
mesh_terms: D000818:Animals; D000917:Antibody Formation; D017931:DNA Primers; D019084:Fluorescent Antibody Technique, Indirect; D005849:Gerbillinae / Q000821:virology*; D051379:Mice; D029262:Puumala virus / Q000254:growth & development* / Q000302:isolation & purification*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014776:Virus Cultivation
chemical_list: D017931:DNA Primers