Tissue culture isolation, electron microscopic characterization and PCR-detection of a betanodavirus isolated from diseased Asian sea bass.
A viral agent was successfully isolated from Asian sea bass juveniles (Letes calcarifer) exhibiting clinical signs of viral nervous necrosis (VNN) in southern Malaysia on February 2008. Eyes and brains of diseased fish were pooled, extracted and filtered. The filtrates were inoculated on SSN-1 cells...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English English |
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2010
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| Online Access: | http://psasir.upm.edu.my/id/eprint/25412/1/Tissue%20culture%20isolation.pdf |
| _version_ | 1796970752609615872 |
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| author | F. Hasoon, Mauida Mohd. Daud, Hassan Arshad, Siti Suri Bejo, Mohd Hair |
| author_facet | F. Hasoon, Mauida Mohd. Daud, Hassan Arshad, Siti Suri Bejo, Mohd Hair |
| author_sort | F. Hasoon, Mauida |
| collection | UPM |
| description | A viral agent was successfully isolated from Asian sea bass juveniles (Letes calcarifer) exhibiting clinical signs of viral nervous necrosis (VNN) in southern Malaysia on February 2008. Eyes and brains of diseased fish were pooled, extracted and filtered. The filtrates were inoculated on SSN-1 cells and incubated at 25°C. Cytopathic effect (CPE) recognized as rounded cells continued to aggregate and the vacuolation. Electron micrographs of the infected SSN-1 cells revealed icosahedral nucleocapsid virions with 22-28 nm in diameter. Viral harvest was resistant to chloroform and iodine treatments, which indicated that it was naked and contained RNA genome. Identification using reverse transcription polymerase chain reaction (RT-PCR) and sequence analysis of PCR product was conducted and gave a single PCR product at 460 bp in agarose gel, followed by nested PCR with specific primers for PCR products at 220 bp. Partial nucleotide sequence of the nervous necrosis virus coat protein gene showed 94.0-96.1% homology to the nucleotide sequences of coat protein gene from nervous necrosis virus isolated from other countries in the Southeast Asia.
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| first_indexed | 2024-03-06T08:02:44Z |
| format | Article |
| id | upm.eprints-25412 |
| institution | Universiti Putra Malaysia |
| language | English English |
| last_indexed | 2024-03-06T08:02:44Z |
| publishDate | 2010 |
| record_format | dspace |
| spelling | upm.eprints-254122015-09-28T02:14:07Z http://psasir.upm.edu.my/id/eprint/25412/ Tissue culture isolation, electron microscopic characterization and PCR-detection of a betanodavirus isolated from diseased Asian sea bass. F. Hasoon, Mauida Mohd. Daud, Hassan Arshad, Siti Suri Bejo, Mohd Hair A viral agent was successfully isolated from Asian sea bass juveniles (Letes calcarifer) exhibiting clinical signs of viral nervous necrosis (VNN) in southern Malaysia on February 2008. Eyes and brains of diseased fish were pooled, extracted and filtered. The filtrates were inoculated on SSN-1 cells and incubated at 25°C. Cytopathic effect (CPE) recognized as rounded cells continued to aggregate and the vacuolation. Electron micrographs of the infected SSN-1 cells revealed icosahedral nucleocapsid virions with 22-28 nm in diameter. Viral harvest was resistant to chloroform and iodine treatments, which indicated that it was naked and contained RNA genome. Identification using reverse transcription polymerase chain reaction (RT-PCR) and sequence analysis of PCR product was conducted and gave a single PCR product at 460 bp in agarose gel, followed by nested PCR with specific primers for PCR products at 220 bp. Partial nucleotide sequence of the nervous necrosis virus coat protein gene showed 94.0-96.1% homology to the nucleotide sequences of coat protein gene from nervous necrosis virus isolated from other countries in the Southeast Asia. 2010 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/25412/1/Tissue%20culture%20isolation.pdf F. Hasoon, Mauida and Mohd. Daud, Hassan and Arshad, Siti Suri and Bejo, Mohd Hair (2010) Tissue culture isolation, electron microscopic characterization and PCR-detection of a betanodavirus isolated from diseased Asian sea bass. Malaysian Applied Biology, 39 (2). pp. 7-13. ISSN 0126-8643 English |
| spellingShingle | F. Hasoon, Mauida Mohd. Daud, Hassan Arshad, Siti Suri Bejo, Mohd Hair Tissue culture isolation, electron microscopic characterization and PCR-detection of a betanodavirus isolated from diseased Asian sea bass. |
| title | Tissue culture isolation, electron microscopic characterization and PCR-detection of a betanodavirus isolated from diseased Asian sea bass. |
| title_full | Tissue culture isolation, electron microscopic characterization and PCR-detection of a betanodavirus isolated from diseased Asian sea bass. |
| title_fullStr | Tissue culture isolation, electron microscopic characterization and PCR-detection of a betanodavirus isolated from diseased Asian sea bass. |
| title_full_unstemmed | Tissue culture isolation, electron microscopic characterization and PCR-detection of a betanodavirus isolated from diseased Asian sea bass. |
| title_short | Tissue culture isolation, electron microscopic characterization and PCR-detection of a betanodavirus isolated from diseased Asian sea bass. |
| title_sort | tissue culture isolation electron microscopic characterization and pcr detection of a betanodavirus isolated from diseased asian sea bass |
| url | http://psasir.upm.edu.my/id/eprint/25412/1/Tissue%20culture%20isolation.pdf |
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