Serotype-specific antigen ELISA for detection of Chiba virus in stools.
Chiba virus (CV), a Norwalk-like virus (NLV), was first identified as a cause of oyster-associated outbreak of gastroenteritis that occurred in Chiba prefecture, Japan, in 1987. An enzyme-linked immunosorbent assay (ELISA), based on hyperimmune antisera to recombinant baculovirus-expressed capsid proteins of CV (rCV), was developed to detect CV antigen in stools. No cross-reactions were observed with other enteric viruses including enteroviruses, rotaviruses, astroviruses, or enteric adenoviruses. The ELISA was used to screen 101 stools collected from 16 oyster-associated outbreaks of acute gastroenteritis. Twelve stools (11.9%) from seven outbreaks were positive for CV antigen. Ten rCV ELISA-positive strains were confirmed by RT-PCR and nucleotide sequencing. ELISA-positive strains showed 96-100% nucleotide sequence identity to each other, though they were obtained nine years apart. Phylogenetic analysis demonstrated that all ten strains clustered with the prototype CV in genogroup I viruses. We concluded that the antigen ELISA described in this study is highly type-specific, and that this method should be useful for epidemiological surveys of Chiba virus infections.
Kobayashi
S
S
Laboratory of Virology, Aichi Prefectural Institute of Public Health, Nagoya, Japan. shinkoba@he.mirai.ne.jp
Sakae
K
K
Natori
K
K
Takeda
N
N
Miyamura
T
T
Suzuki
Y
Y
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002139:Caliciviridae / Q000276:immunology / Q000302:isolation & purification*; D017250:Caliciviridae Infections / Q000175:diagnosis / Q000821:virology*; D002213:Capsid / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005243:Feces / Q000821:virology*; D005759:Gastroenteritis / Q000175:diagnosis / Q000821:virology*; D006168:Guinea Pigs; D007564:Japan; D008969:Molecular Sequence Data; D010125:Ostreidae / Q000821:virology; D010802:Phylogeny; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D011994:Recombinant Proteins