Expression of recombinant capsid proteins of chitta virus, a genogroup II Norwalk virus, and development of an ELISA to detect the viral antigen.
The second open reading frame (ORF2) gene of the Chitta virus (CHV) was cloned to construct a recombinant baculovirus. The CHV ORF2 is predicted to encode a capsid protein of 535 amino acids (aa). CHV showed a high aa identity in the capsid region with genogroup II Norwalk virus (NV) (65-85%), but a low aa identity with genogroup I NV (44-46%). Phylogenetic analysis of the ORF2 gene demonstrated that CHV is genetically closely related to the Hawaii virus included in genogroup II NV. The recombinant capsid protein of CHV (rCHV) self-assembled to form empty virus-like particles (VLPs) when expressed in insect cells with the recombinant baculovirus. An enzyme-linked immunosorbent assay (ELISA) based on antisera to rCHV was developed to detect CHV antigen in stools. The antigen ELISA appeared to be highly specific to both rCHV and CHV-like strains. In addition, combined use of antigen ELISAs using antibodies against two antigenically distinct recombinant VLPs, the recombinant Chiba virus (rCV) and recombinant Seto virus (rSEV), enabled us to determine the genetic as well as antigenic relationship among these three viruses.
Kobayashi
S
S
Development Department, Mitsubishi Kagaku Bio-Clinical Laboratories Inc., Tokyo, Japan. shinkoba@he.mirai.ne.jp
Sakae
K
K
Suzuki
Y
Y
Ishiko
H
H
Kamata
K
K
Suzuki
K
K
Natori
K
K
Miyamura
T
T
Takeda
N
N
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis*; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D036022:Capsid Proteins; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005243:Feces / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D009663:Norwalk virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D010802:Phylogeny; D011994:Recombinant Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D018411:Spodoptera / Q000821:virology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; C121877:Norwalk virus capsid; D011994:Recombinant Proteins