Korean Journal of Fisheries and Aquatic Sciences (한국수산과학회지)

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
권호 표지
DOI QR코드

DOI QR Code

Detection and Genetic Differentiation of Megalocytiviruses in Shellfish, via High-Resolution Melting (HRM) Analysis

HRM 분석법을 이용한 패류 내 Megalocytiviruses의 검출과 유전적 분석

  • Kim, Kwang Il (Aquatic Life Disease Control Division, National Fisheries Research & Development Institute) ;
  • Jin, Ji Woong (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Kim, Young Chul (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Jeong, Hyun Do (Department of Aquatic Life Medicine, Pukyong National University)
  • 김광일 (국립수산과학원 수산생물방역과) ;
  • 진지웅 (부경대학교 수산생명의학과) ;
  • 김영철 (부경대학교 수산생명의학과) ;
  • 정현도 (부경대학교 수산생명의학과)
  • Received : 2014.04.21
  • Accepted : 2014.06.16
  • Published : 2014.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Viruses in the genus Megalocytivirus have been subdivided into four subgroups. Among these subgroups 2 and 4, represented by the red sea bream iridovirus (RBIV) and the olive flounder iridovirus (FLIV), respectively, are non-exotic. subgroups 1 and 3, represented by the red sea bream iridovirus (RSIV) and the infectious spleen and kidney necrosis virus (ISKNV), respectively, have not been detected in Korea and are known as exotic. Shellfish are filter-feeders, and can thus filter and accumulate Megalocytivirus in their digestive glands, allowing us to track viral contamination in surrounding aquatic environment. In this study, we developed a high-resolution melting (HRM) analysis to differentiate among subgroups of Megalocytivirus accumulated in shellfish, and confirmed the convenience and efficiency of this method. More than two subgroups of Megalocytivirus were found in the digestive gland of a single shellfish. We classified all Megalocytivirus viruses from shellfish in Korea into subgroups 2 and 4, although proportions of subgroups were different among regions. Compared to nucleotide sequencing analysis, HRM analysis is a simple and rapid method for differentiating of Megalocytivirus subgroups.

Keywords

References

  1. Atmar RL, Neill FH, Romalde JL, Le Guyader F, Woodley CM, Metcalf TG and Estes MK. 1995. Detection of Norwalk virus and hepatitis A viruses in shellfish tissues with the PCR. Appl Environ Microbiol 61, 3014-3018.
  2. Do JW, Cha SJ, Kim JS, Jeong EJ, An EJ, Lee NS, Choi HJ, Lee CH, Park MS, Kim JW, Kim YC and Park JW. 2005. Phylogenetic analysis of the major capsid protein gene of iridovirus isolates from cultured flounders Paralichthys olivaceus in Korea. Dis Aquat Organ 64, 193-200. http://dx.doi.org/10.3354/dao064193.
  3. Gerba CP and Goyal SM. 1978. Detection and occurrence of enteric viruses in shellfish: A review. J Food Prot 41, 743-754. https://doi.org/10.4315/0362-028X-41.9.743
  4. Goyal SM, Gerba CP and Melnick JL. 1979. Human enteroviruses in oysters and their overlying waters. Appl Environ Microbiol 37, 572-575.
  5. Inouye K, Yamano K, Maeno Y, Nakajima K, Matsuoka M, Wada Y and Sorimachi M. 1992. Iridovirus infection of cultured red sea bream, Pagrus major. Fish Pathol 27, 19-27. https://doi.org/10.3147/jsfp.27.19
  6. Imajoh M, Ikawa T and Oshima SI. 2007. Characterization of a new fibroblast cell line from a tail fin of red sea bream, Pagrus major, and phylogenetic relationships of a recent RSIV isolate in Japan. Virus Res 126, 45-52. http://dx.doi.org/10.1016/j.virusres.2006.12.020.
  7. Jeong JB, Jun LJ, Yoo MH, Kim MS, Komisar JL and Jeong HD. 2003. Characterization of the DNA nucleotide sequences in the genome of red sea bream iridoviruses isolated in Korea. Aquaculture 220, 119-133. https://doi.org/10.1016/S0044-8486(02)00538-0
  8. Jeong JB, Kim HY, Jun LJ, Lyu JH, Park NG, Kim JK and Jeong HD. 2008. Outbreaks and risks of infectious spleen and kidney necrosis virus disease in freshwater ornamental fishes. Dis Aquat Organ 78, 209-215 https://doi.org/10.3354/dao01879
  9. Kim JW, Cho MY, Jin JW, Kim KH, Kim KI and Jeong HD. 2011. Detection of Megalocytivirus in shellfish using PCR with various DNA extraction methods. J Fish Pathol 24, 65-73. https://doi.org/10.7847/jfp.2011.24.2.065
  10. Kurita J and Nakajima K. 2012. Megalocytiviruses. Viruses 4, 521-538. https://doi.org/10.3390/v4040521
  11. Kristensen LS and Dobrovic A. 2008. Direct genotyping of single nucleotide polymorphisms in methyl metabolism genes using probe-free high-resolution melting analysis. Cancer Epidemiol Biomarkers Prev 17, 1240-1247. http://dx.doi.org/10.1158/1055-9965.EPI-07-2531.
  12. Oh MJ, Kitamura SI, Kim WS, Park MK, Jung SJ, Miyadai T and Ohtani M. 2006. Susceptibility of marine fish species to a Megalocytivirus, turbot iridovirus, isolated from turbot, Psetta maximus (L.). J Fish Dis 29, 415-421. https://doi.org/10.1111/j.1365-2761.2006.00734.x
  13. Suzuki S and Nojima M. 1999. Detection of a marine birnavirus in wild molluscan shellfish species from Japan. Fish Pathol 34, 121-125. https://doi.org/10.3147/jsfp.34.121
  14. Tajiri-Utagawa E, Hara M, Takahashi K, Watanabe M and Wakita T. 2008. Development of a rapid high-throughput method for high-resolution melting analysis for routine detection and genotyping of noroviruses. J Clin Microbiol 47, 435-440. http://dx.doi.org/10.1128/JCM.01247-08.
  15. Vazquez-Boucard C, Alvarez-Ruiz P, Escobedo-Fregoso C, Anguiano-Vega G, Duran-Avelar Mde J, Pinto VS and Escobedo- Bonilla CM. 2010. Detection of white spot syndrome virus (WSSV) in the Pacific oyster Crassostrea gigas. J Invertebr Pathol 104, 245-247. https://doi.org/10.1016/j.jip.2010.04.004
  16. Wittwer CT, Reed GH, Gundry CN, Vandersteen JG and Pryor RJ. 2003. High-resolution genotyping by amplicon melting analysis using LCGreen. Am Assoc Clin Chem 49, 853-860. https://doi.org/10.1373/49.6.853

Cited by

  1. vol.41, pp.8, 2018, https://doi.org/10.1111/jfd.12815