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Development of a stringent ELISA protocol to evaluate anti-viral hemorrhagic septicemia virus-specific antibodies in olive flounder Paralichthys olivaceus with improved specificity
Hyoung Jun Kim , Jeong Su Park , Se Ryun Kwon
J. Microbiol. 2015;53(7):481-485.   Published online June 27, 2015
DOI: https://doi.org/10.1007/s12275-015-5101-9
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AbstractAbstract
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.

Citations

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  • Development of an indirect ELISA for detection of the adaptive immune response of black carp (Mylopharyngodon piceus)
    Tongtong Wang, Shanshan Jin, Ruoxuan Lv, Yuting Meng, Guozhong Li, Yuxing Han, Qiusheng Zhang
    Journal of Immunological Methods.2023; 521: 113550.     CrossRef
  • Detection of Salmonid IgM Specific to the Piscine Orthoreovirus Outer Capsid Spike Protein Sigma 1 Using Lipid-Modified Antigens in a Bead-Based Antibody Detection Assay
    Lena Hammerlund Teige, Subramani Kumar, Grethe M. Johansen, Øystein Wessel, Niccolò Vendramin, Morten Lund, Espen Rimstad, Preben Boysen, Maria K. Dahle
    Frontiers in Immunology.2019;[Epub]     CrossRef
  • Production of a monoclonal antibody against of muskellunge (Esox masquinongy) IgM heavy chain and its use in development of an indirect ELISA for titrating circulating antibodies against VHSV-IVB
    Mohamed Faisal, Isaac F. Standish, Mary Ann Vogelbein, Elena V. Millard, Stephen L. Kaattari
    Fish & Shellfish Immunology.2019; 88: 464.     CrossRef
  • Phylogenetic analysis and duplex RT-PCR detection of viral hemorrhagic septicemia virus in olive flounder (Paralichthys olivaceus) from Korea
    Jee Youn Hwang, Seongdo Lee, Thanthrige Thiunuwan Priyathilaka, Hyerim Yang, Hyukjae Kwon, Mun Gyeong Kwon, Seong Don Hwang, Myoung-Jin Kim, Jehee Lee
    Aquaculture.2018; 484: 242.     CrossRef
  • Herpesvirus Infection Induces both Specific and Heterologous Antiviral Antibodies in Carp
    Julio M. Coll
    Frontiers in Immunology.2018;[Epub]     CrossRef
  • Validation of a serum neutralization test for detection of antibodies specific to cyprinid herpesvirus 3 in infected common and koi carp (Cyprinus carpio)
    J Cabon, L Louboutin, J Castric, S Bergmann, G Bovo, M Matras, O Haenen, N J Olesen, T Morin
    Journal of Fish Diseases.2017; 40(5): 687.     CrossRef
  • The amino-terminal domain of ORF149 of koi herpesvirus is preferentially targeted by IgM from carp populations surviving infection
    F. Torrent, A. Villena, P. A. Lee, W. Fuchs, S. M. Bergmann, J. M. Coll
    Archives of Virology.2016; 161(10): 2653.     CrossRef
Distribution of Marine Birnavirus in Cultured Olive Flounder Paralichthys olivaceus in Korea
Sung-Ju Jung , Seok-Ryel Kim , Il-Yong Joung , Shin-Ichi Kitamura , Hee-Taek Ceong , Myung-Joo Oh
J. Microbiol. 2008;46(3):265-273.   Published online July 5, 2008
DOI: https://doi.org/10.1007/s12275-008-0004-7
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AbstractAbstract
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
Seong-Joon Joh , Jeong-Ho Kim , Gang-Joon Heo
J. Microbiol. 2000;38(1):44-47.
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AbstractAbstract
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.

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