Generation of monoclonal antibodies specific for ORF68 of koi herpesvirus.
Outbreaks of koi herpesvirus (KHV) infection in carp are still a serious problem worldwide. KHV is closely related to other two cyprinid herpesviruses, pox herpesvirus (CHV) and haematopoietic necrosis herpesvirus (CyHV-2) in goldfish. In this study, two major KHV antigenic proteins (ORF62 and ORF68) were identified by immunoscreening using a KHV-specific polyclonal antibody, and then monoclonal antibodies were generated for immunodiagnostic studies. After screening hybridoma cells, one mAb against ORF68 (mAb-7C6) was obtained but no mAbs against ORF62. mAb-7C6 specifically reacted with a lysate of KHV-infected koi fin cells (KF-1 cells) but not with lysates of CHV- or CyHV-2-infected KF-1 cells in an immuno-blotting analysis. Similar results were shown in the following tests: (1) a indirect fluorescent antibody test using infected KF-1 cells and (2) an immunohistochemical investigation by fast red stain (infected liver) or FITC detection (infected spleen). These results suggested that mAb-7C6 specifically reacts with KHV ORF68 protein.
Laboratory of Genome Science, Tokyo University of Marine Science and Technology, 4-5-7 Konan, Minato, Tokyo 108-8477, Japan. aoki@kaiyodai.ac.jp
Takashi
T
Aoki
Tomokazu
T
Takano
Sasimnanas
S
Unajak
Madoka
M
Takagi
Young Rim
YR
Kim
Seong Bin
SB
Park
Hidehiro
H
Kondo
Ikuo
I
Hirono
Tatsuo
T
Saito-Taki
Jun-Ichi
J
Hikima
Tae Sung
TS
Jung
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D002347:Carps / Q000821:virology; D002460:Cell Line; D005393:Fish Diseases / Q000175:diagnosis* / Q000276:immunology; D006564:Herpesviridae / Q000235:genetics* / Q000276:immunology; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D011994:Recombinant Proteins