Antigenic properties of porcine teschovirus 1 (PTV-1) Talfan strain and molecular strategy for serotyping of PTVs.
For reliable diagnosis of porcine teschovirus (PTV) infection we created an RT-PCR-based molecular strategy for serotyping that encompassed the dominant neutralizing antigenic site of PTV, followed by phylogenetic analyses of amplicons. We identified neutralizing antigenic sites of PTV-1 Talfan strain through epitope mapping of neutralizing monoclonal antibodies (MAbs), using synthetic peptides spanning the capsid proteins. All 11 MAbs obtained recognized peptides in the EF loop ("puff") of VP2 protein. Two MAbs concurrently reacted to peptides, one in the GH loop of VP1 and one in the VP1 C terminus. Three-dimensional modeling of Talfan capsid protein predicted exposure of all these sites on the virion surface in a close line centered around puff. We then designed a single pair of degenerate primers to VP2 and amplified the region of approximately 320 bp encompassing puff in 8 PTV prototype strains and 6 field isolates. Phylogenetic analyses of the puff sequences of 11 prototype strains and 34 field isolates obtained from databanks showed that all homotypic strains (both field and prototype) were always monophyletic, except for one 'untypable' Japanese strain. This RT-PCR-based strategy appears to be a reliable surrogate for serotyping and could facilitate the diagnosis and epidemiological study of PTV infection.
Kaku
Y
Y
Department of Exotic Diseases, National Institute of Animal Health, Tokyo, Japan. ykaku@nih.go.jp
Murakami
Y
Y
Sarai
A
A
Wang
Y
Y
Ohashi
S
S
Sakamoto
K
K
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D017931:DNA Primers; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009500:Neutralization Tests / Q000379:methods*; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D010802:Phylogeny; D010850:Picornaviridae Infections / Q000175:diagnosis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012703:Serotyping / Q000379:methods*; D052700:Teschovirus / Q000145:classification* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D017931:DNA Primers; D000939:Epitopes; D010455:Peptides