Multiplex PCR method for distinguishing two genotypes of Megalocytivirus isolates in Korea
Megalocytivirus is a genus of piscine viruses that belongs to the Iridoviridae family, and this family includes red seabream iridovirus (RSIV), turbot reddish body iridovirus (TRBIV), and infectious spleen and kidney necrosis virus. RSIV causes high mortality and economic losses in the rock bream (O...
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ftdoajarticles:oai:doaj.org/article:8150897fda724d02b893ad5b93a4373e 2023-05-15T18:41:11+02:00 Multiplex PCR method for distinguishing two genotypes of Megalocytivirus isolates in Korea Hanchang Sohn Seongdo Lee Hyukjae Kwon Mun Gyeong Kwon Jee Youn Hwang Seong Don Hwang Jehee Lee 2021-11-01T00:00:00Z https://doi.org/10.1016/j.aqrep.2021.100886 https://doaj.org/article/8150897fda724d02b893ad5b93a4373e EN eng Elsevier http://www.sciencedirect.com/science/article/pii/S2352513421003021 https://doaj.org/toc/2352-5134 2352-5134 doi:10.1016/j.aqrep.2021.100886 https://doaj.org/article/8150897fda724d02b893ad5b93a4373e Aquaculture Reports, Vol 21, Iss , Pp 100886- (2021) Megalocytivirus Red seabream iridovirus Turbot reddish body iridovirus Multiplex polymerase chain reaction Aquaculture. Fisheries. Angling SH1-691 article 2021 ftdoajarticles https://doi.org/10.1016/j.aqrep.2021.100886 2022-12-31T13:36:58Z Megalocytivirus is a genus of piscine viruses that belongs to the Iridoviridae family, and this family includes red seabream iridovirus (RSIV), turbot reddish body iridovirus (TRBIV), and infectious spleen and kidney necrosis virus. RSIV causes high mortality and economic losses in the rock bream (Oplegnathus fasciatus) Korean aquaculture industry. The World Organization for Animal Health’s Office International des Epizooties has provided a manual for RSIV detection. However, distinguishing it from TRBIV has not been confirmed. In this study, a multiplex PCR method was established to detect two genotypes of Megalocytivirus, RSIV and TRBIV. New primer pairs were optimized for the PCR reaction. A mixture of one universal and two specific primer pairs could amplify three distinct products targeting ATPase, DPO, and LRP genes. The sensitivities of primer pairs were evaluated, showing a detection limit of 2.0 × 105 copies, using a plasmid construct, against the three target genes of Megalocytivirus. Moreover, specificity evaluation indicated that primer pairs were not subjected to interference by other virus strains. This evaluation of multiplex PCR showed that it can distinguish two genotypes of Megalocytivirus from 21 Korean Megalocytivirus isolates, including 20 RSIV and one TRBIV. Finally, we describe a new multiplex PCR method to detect different genotypes of Megalocytivirus simultaneously, which makes the diagnosis of viral diseases occurring in the Korean aquaculture industry more convenient. Article in Journal/Newspaper Turbot Directory of Open Access Journals: DOAJ Articles Aquaculture Reports 21 100886 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Megalocytivirus Red seabream iridovirus Turbot reddish body iridovirus Multiplex polymerase chain reaction Aquaculture. Fisheries. Angling SH1-691 |
spellingShingle |
Megalocytivirus Red seabream iridovirus Turbot reddish body iridovirus Multiplex polymerase chain reaction Aquaculture. Fisheries. Angling SH1-691 Hanchang Sohn Seongdo Lee Hyukjae Kwon Mun Gyeong Kwon Jee Youn Hwang Seong Don Hwang Jehee Lee Multiplex PCR method for distinguishing two genotypes of Megalocytivirus isolates in Korea |
topic_facet |
Megalocytivirus Red seabream iridovirus Turbot reddish body iridovirus Multiplex polymerase chain reaction Aquaculture. Fisheries. Angling SH1-691 |
description |
Megalocytivirus is a genus of piscine viruses that belongs to the Iridoviridae family, and this family includes red seabream iridovirus (RSIV), turbot reddish body iridovirus (TRBIV), and infectious spleen and kidney necrosis virus. RSIV causes high mortality and economic losses in the rock bream (Oplegnathus fasciatus) Korean aquaculture industry. The World Organization for Animal Health’s Office International des Epizooties has provided a manual for RSIV detection. However, distinguishing it from TRBIV has not been confirmed. In this study, a multiplex PCR method was established to detect two genotypes of Megalocytivirus, RSIV and TRBIV. New primer pairs were optimized for the PCR reaction. A mixture of one universal and two specific primer pairs could amplify three distinct products targeting ATPase, DPO, and LRP genes. The sensitivities of primer pairs were evaluated, showing a detection limit of 2.0 × 105 copies, using a plasmid construct, against the three target genes of Megalocytivirus. Moreover, specificity evaluation indicated that primer pairs were not subjected to interference by other virus strains. This evaluation of multiplex PCR showed that it can distinguish two genotypes of Megalocytivirus from 21 Korean Megalocytivirus isolates, including 20 RSIV and one TRBIV. Finally, we describe a new multiplex PCR method to detect different genotypes of Megalocytivirus simultaneously, which makes the diagnosis of viral diseases occurring in the Korean aquaculture industry more convenient. |
format |
Article in Journal/Newspaper |
author |
Hanchang Sohn Seongdo Lee Hyukjae Kwon Mun Gyeong Kwon Jee Youn Hwang Seong Don Hwang Jehee Lee |
author_facet |
Hanchang Sohn Seongdo Lee Hyukjae Kwon Mun Gyeong Kwon Jee Youn Hwang Seong Don Hwang Jehee Lee |
author_sort |
Hanchang Sohn |
title |
Multiplex PCR method for distinguishing two genotypes of Megalocytivirus isolates in Korea |
title_short |
Multiplex PCR method for distinguishing two genotypes of Megalocytivirus isolates in Korea |
title_full |
Multiplex PCR method for distinguishing two genotypes of Megalocytivirus isolates in Korea |
title_fullStr |
Multiplex PCR method for distinguishing two genotypes of Megalocytivirus isolates in Korea |
title_full_unstemmed |
Multiplex PCR method for distinguishing two genotypes of Megalocytivirus isolates in Korea |
title_sort |
multiplex pcr method for distinguishing two genotypes of megalocytivirus isolates in korea |
publisher |
Elsevier |
publishDate |
2021 |
url |
https://doi.org/10.1016/j.aqrep.2021.100886 https://doaj.org/article/8150897fda724d02b893ad5b93a4373e |
genre |
Turbot |
genre_facet |
Turbot |
op_source |
Aquaculture Reports, Vol 21, Iss , Pp 100886- (2021) |
op_relation |
http://www.sciencedirect.com/science/article/pii/S2352513421003021 https://doaj.org/toc/2352-5134 2352-5134 doi:10.1016/j.aqrep.2021.100886 https://doaj.org/article/8150897fda724d02b893ad5b93a4373e |
op_doi |
https://doi.org/10.1016/j.aqrep.2021.100886 |
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Aquaculture Reports |
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21 |
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100886 |
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