Research Support, Non-U.S. Gov'ts
- Development of a stringent ELISA protocol to evaluate anti-viral hemorrhagic septicemia virus-specific antibodies in olive flounder Paralichthys olivaceus with improved specificity
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Hyoung Jun Kim , Jeong Su Park , Se Ryun Kwon
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J. Microbiol. 2015;53(7):481-485. Published online June 27, 2015
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DOI: https://doi.org/10.1007/s12275-015-5101-9
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Abstract
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Olive flounder were vaccinated with polyinosinic:polycytidylic
acid [Poly (I:C)] to prevent viral hemorrhagic septicemia
(VHS). Vaccine efficacy was verified by detection of anti-
VHS virus (VHSV) antibodies using enzyme-linked immunosorbent
assay (ELISA). In the study, ELISA absorbance values
of the negative control group [Poly (I:C)-MEM10] were saturated
when an ELISA protocol, that includes pretreatment
of the fish sera with 5% skim milk, was used. However,
the saturated OD values in the negative control did not correlate
with a specific immune response against VHSV, because
the group showed low survival rate (only 10%) following
the VHSV challenge. Also, OD values of Poly (I:C)-
VHSV group were high, and the group showed high survival
rate (97.5%) against VHSV challenge test. It was suggested
that the high OD values were possibly due to the
presence of anti-fetal bovine serum (FBS) cross-reactivity.
To compensate this, we subtracted the absorbance of infectious
hematopoietic necrosis (IHNV)-Ag plates from
those of the VHSV-Ag plates. However, the average value
for the Poly (I:C)-VHSV group (0.167) was lower than expected
even though high survival rate. We used an advanced
ELISA system to pre-treat fish sera with 5% skim milk and
two novirhabdoviruses as capture antigens as well as 50%
FBS. The corrected absorbance values for pre-treated fish
sera from the negative control Poly (I:C)-MEM10 and experimental
Poly (I:C)-VHSV groups averaged 0.033 and
0.579, respectively. The specific VHSV antibody response
of the vaccinated group was assessed using fish sera pretreated
with skim milk and FBS and by calculating the corrected
absorbance values from ELISA with two novirhabdovirus
capture antigens.
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Citations
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- Distribution of Marine Birnavirus in Cultured Olive Flounder Paralichthys olivaceus in Korea
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Sung-Ju Jung , Seok-Ryel Kim , Il-Yong Joung , Shin-Ichi Kitamura , Hee-Taek Ceong , Myung-Joo Oh
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J. Microbiol. 2008;46(3):265-273. Published online July 5, 2008
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DOI: https://doi.org/10.1007/s12275-008-0004-7
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Scopus
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Abstract
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Surveys of marine birnavirus (MABV) were undertaken in cultured olive flounder Paralichthys olivaceus from the south and west coastal areas and Jeju in Korea during the period January 1999 to April 2007. MABV was detected in all seasons from the fry, juveniles and adult fish from the areas examined. Evident cytopathic effects of the virus including rounding and cell lysis were observed in chinook salmon embryo (CHSE-214) and rainbow trout gonad (RTG-2) cells, but not in fathead minnow (FHM) and epithelial papilloma of carp (EPC) cells. Nucleotide sequences of the VP2/NS junction region of the Korean isolates showed 97.8%~100% similarity, and they belonged to the same genogroup. Cross neutralization tests with serotype-specific rabbit antisera against MABV strains exhibited a close antigenic relationships between strains, and were distinct from infectious pancreatic necrosis virus (IPNV) strains. Coinfection of MABV with bacteria (Streptococcus iniae, Vibrio spp.) and viruses (nervous necrosis virus, lymphocystis disease virus,
viral hemorrhagic septicemia virus) was observed.
- Genetic Analysis of the VP2/NS Junction Region on Segment A of Marine Birnavirus Isolated from Cultured Flounder Paralichthys olivaceous
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Seong-Joon Joh , Jeong-Ho Kim , Gang-Joon Heo
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J. Microbiol. 2000;38(1):44-47.
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Abstract
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The cDNA of VP2/NS junction region on segment A of the marine birnavirus (MBV) isolated from flounder (DS strain) was amplified using the reverse transcription (RT)-polymerase chain reaction (PCR). Its cDNA nucleotide and deduced amino acid sequences were analyzed, and compared with the reference strains of MBV and infectious pancreatic necrosis virus (IPNV). Analyses of the nucleotide and deduced amino acid sequences revealed that the DS strain is very similar to the reference strains of MBV, distant from the IPNV.