Efficient detection of pathogen virus in sand dabs, <i>Paralichthys olivaceus</i> using loop-mediated isothermal amplification (LAMP)

HWANG Jinik; PARK So Yun; SUH Sung-Suk; PARK Mirye; LEE Sukchan; LEE Taek-Kyun

doi:10.1007/s13131-016-0889-7

Article Contents

Article Navigation > Acta Oceanologica Sinica > 2016 > 35(8): 44-50

HWANG Jinik, PARK So Yun, SUH Sung-Suk, PARK Mirye, LEE Sukchan, LEE Taek-Kyun. Efficient detection of pathogen virus in sand dabs, Paralichthys olivaceus using loop-mediated isothermal amplification (LAMP)[J]. Acta Oceanologica Sinica, 2016, 35(8): 44-50. doi: 10.1007/s13131-016-0889-7

Citation:

HWANG Jinik, PARK So Yun, SUH Sung-Suk, PARK Mirye, LEE Sukchan, LEE Taek-Kyun. Efficient detection of pathogen virus in sand dabs, Paralichthys olivaceus using loop-mediated isothermal amplification (LAMP)[J]. Acta Oceanologica Sinica, 2016, 35(8): 44-50. doi: 10.1007/s13131-016-0889-7

Citation:

PDF( 17616 KB)

Efficient detection of pathogen virus in sand dabs, Paralichthys olivaceus using loop-mediated isothermal amplification (LAMP)

doi: 10.1007/s13131-016-0889-7

1.
South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje 53201, Republic of Korea;Korea University of Science and Technology, Daejeon 34113, Republic of Korea
2.
South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje 53201, Republic of Korea
3.
Department of Genetic Engineering, Sungkyunkwan University, Suwon 16419, Republic of Korea

Received Date: 2015-08-11
Rev Recd Date: 2015-12-16

Abstract

Abstract

Viral hemorrhagic septicemia virus (VHSV) and marine birnavirus (MABV) are the causative pathogens for some of the most explosive epidemics of emerging viral diseases in many Asian countries, leading to huge economic losses in aquaculture. Rapid molecular detection for surveillance or diagnosis has been a critical component in reducing the prevalence of pathogen infection. The loop-mediated isothermal amplification (LAMP) of DNA is currently one of the most commonly used molecular diagnostic tools, as it is simple, quick, and easy to amplify target DNA under isothermal conditions. In the present study, a novel and highly specific LAMP assay for the sensitive and rapid detection of VHSV and MABV infection in fish was developed. Using a set of synthesized primers matching a specific region of the genome, the efficiency and specificity of the LAMP assay were optimized in terms of the reaction temperature and DNA polymerase concentration, as they are the main determinants of the sensitivity and specificity of the LAMP assay. In particular, we demonstrated that our assay could be applied to efficient detection of VHSV and MABV infection in the wild fish, Paralichthys olivaceus. Our results demonstrate the simplicity and convenience of this method for the detection of viral infection in aquatic organisms.
- viral hemorrhagic septicaemia virus (VHSV),
- marine birnavirus (MABV),
- polymerase chain reaction,
- loop-mediated isothermal amplification

FullText(HTML)

References(29)

References

Al-Hussinee L, Lumsden J S. 2011. Detection of VHSV IVb within the gonads of Great Lakes fish using in situ hybridization. Dis Aquat Organ, 95(1):81-86

Azumi K, Nakamura S, Kitamura S I, et al. 2007. Accumulation of or-ganotin compounds and marine birnavirus detection in Korean ascidians. Fish Sci, 73(2):263-269

Bolasina S, Tagawa M, Yamashita Y, et al. 2006. Effect of stocking density on growth, digestive enzyme activity and cortisol level in larvae and juveniles of Japanese flounder, Paralichthys oli-vaceus. Aquaculture, 259(1-4):432-443

Caipang C M, Hirono I, Aoki T, 2003. Development of a real-time PCR assay for the detection and quantification of red seabream iridovirus (RSIV). Fish Pathol (in Japanese), 38(1):1-7

Chinchar V G, Hyatt A, Miyazaki T, et al. 2009. Family Iridoviridae:poor viral relations no longer. In:Van Etten J L, ed. Lesser Known Large dsDNA Viruses. Berlin Heidelberg:Springer, 123-170

Dhar A K, Roux M M, Klimpel K R. 2001. Detection and quantifica-tion of infectious hypodermal and hematopoietic necrosis vir-us and white spot virus in shrimp using real-time quantitative PCR and SYBR Green chemistry. J Clin Microbiol, 39(8):2835-2845

Go J, Lancaster M, Deece K, et al. 2006. The molecular epidemiology of iridovirus in Murray cod (Maccullochella peelii peelii) and dwarf gourami (Colisa lalia) from distant biogeographical re-gions suggests a link between trade in ornamental fish and emerging iridoviral diseases. Mol Cell Probes, 20(3-4):212-222

Hirayama T, Imajoh M, Oshima S I. 2005. Quantification of aquabirnaviruses isolated from different host species by real-time RT-PCR. Microbiol Immunol, 49(4):361-371

Hoffmann B, Beer M, Schütze H, et al. 2005. Fish rhabdoviruses:mo-lecular epidemiology and evolution. In:Fu Z F, ed. The World of Rhabdoviruses. Berlin Heidelberg:Springer, 81-117

Hope K M, Casey R N, Groocock G H, et al. 2010. Comparison of quantitative RT-PCR with cell culture to detect viral hemor-rhagic septicemia virus (VHSV) IVb infections in the Great Lakes. J Aquat Anim Health, 22(1):50-61

Huang S M, Tu Chen, Tseng C H, et al. 2011. Genetic analysis of fish iridoviruses isolated in Taiwan during 2001-2009. Arch Virol, 156(9):1505-1515

Inaba M, Kimura T, Kikukawa R, et al. 2007. Annual dynamics of mar-ine birnavirus (MABV) in cultured Japanese flounder Paralich-thys olivaceus and sea water. Fish Sci, 73(3):615-622

Inaba M, Suzuki S, Kitamura S I, et al. 2009. Distribution of marine birnavirus (MABV) in marine organisms from Okinawa, Japan, and a unique sequence variation of the VP2/NS region. J Micro-biol, 47(1):76-84

Jeong J B, Kim H Y, Kim K H, et al. 2006. Molecular comparison of iridoviruses isolated from marine fish cultured in Korea and imported from China. Aquaculture, 255(1-4):105-116

Kamakura M, Suzuki S, Kusuda R. 1995. Characterization of birnavir-us isolated from diseased tiger puffer. In:Program and Ab-stracts of the Autumn Meeting of the Japanese Society of Fish Pathology (in Japanese). Ise, Japan:Japanese Society of Fish Pathology, 14

Kitamura S I, Kamata S I, Nakano S I, et al. 2003. Detection of marine birnavirus genome in zooplankton collected from the Uwa Sea, Japan. Dis Aquat Organ, 54(1):69-72

Kitamura S I, Ko J Y, Lee W L, et al. 2007. Seasonal prevalence of lymphocystis disease virus and aquabirnavirus in Japanese flounder, Paralichthys olivaceus and blue mussel, Mytilus gallo-provincialis. Aquaculture, 266(1-4):26-31

Kuboki N, Inoue N, Sakurai T, et al. 2003. Loop-mediated isothermal amplification for detection of African trypanosomes. J Clin Mi-crobiol, 41(12):5517-5524

Kurita J, Nakajima K, Hirono I, et al. 1998. Polymerase chain reaction (PCR) amplification of DNA of red sea bream[Pagrus major] iridovirus (RSIV). Fish Pathol (in Japanese), 33:17-23

Lang A S, Rise M L, Culley A I, et al. 2009. RNA viruses in the sea. FEMS Microbiol Rev, 33(2):295-323

Leong J C, Brown D, Dobos P, et al. 2000. Family Birnaviridae. Virus taxonomy:Classification and Nomenclature of Viruses. In:Van Regenmortel M H V, Fauquet C M, Bishop D H L, et al., eds. Seventh report of the International Committee on Taxonomy of Viruses. San Diego:Academic, 481-490

Liu Wangta, Hsu C H, Chang C Y, et al. 2006. Immune response against grouper nervous necrosis virus by vaccination of virus-like particles. Vaccine, 24(37-39):6282-6287

Mori Y, Nagamine K, Tomita N, et al. 2001. Detection of loop-medi-ated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem and Bio-phys Res Commun, 289(1):150-154

Nakajima K, Sorimachi M. 1995. Production of monoclonal antibod-ies against red sea bream[Pagrus major] iridovirus. Fish Pathol (in Japanese), 30:47-52

Notomi T, Okayama H, Masubuchi H, et al. 2000. Loop-mediated iso-thermal amplification of DNA. Nucl Acids Res, 28(12):e63-e63

Parida M, Sannarangaiah S, Dash P K, et al. 2008. Loop mediated iso-thermal amplification (LAMP):a new generation of innovative gene amplification technique; perspectives in clinical diagnos-is of infectious diseases. Rev Med Virol, 18(6):407-421

Takano R, Nishizawa T, Arimoto M, et al. 2000. Isolation of viral hem-orrhagic septicemia virus (VHSV) from wild Japanese flounder, Paralichthys olivaceus. Bull Eur Assoc Fish Pathol, 20:186-193

Williams T, Barbosa-Solomieu V, Chinchar V G. 2005. A decade of ad-vances in iridovirus research. Adv Virus Res, 65:173-248

Wolf K. 1988. Fish Viruses and Fish Viral Diseases. Ithaca, NY:Cor-nell University Press

Relative Articles

Supplements(0)

Cited By

Proportional views