Journal of Veterinary Science

The Korean Society of Veterinary Science (대한수의학회)
권호 표지
DOI QR코드

DOI QR Code

Rapid detection of deformed wing virus in honeybee using ultra-rapid qPCR and a DNA-chip

  • Kim, Jung-Min (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Lim, Su-Jin (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Kim, SoMin (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Kim, MoonJung (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Kim, ByoungHee (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Tai, Truong A (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Kim, Seonmi (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Yoon, ByoungSu (Department of Life Science, College of Fusion Science, Kyonggi University)
  • Received : 2019.02.12
  • Accepted : 2019.09.24
  • Published : 2020.01.31
Download PDF
⟨ Previous Next ⟩

Abstract

Fast and accurate detection of viral RNA pathogens is important in apiculture. A polymerase chain reaction (PCR)-based detection method has been developed, which is simple, specific, and sensitive. In this study, we rapidly (in 1 min) synthesized cDNA from the RNA of deformed wing virus (DWV)-infected bees (Apis mellifera), and then, within 10 min, amplified the target cDNA by ultra-rapid qPCR. The PCR products were hybridized to a DNA-chip for confirmation of target gene specificity. The results of this study suggest that our method might be a useful tool for detecting DWV, as well as for the diagnosis of RNA virus-mediated diseases on-site.

Keywords

Acknowledgement

This work (Grants No. 318093-03) was supported by Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry(IPET) through Agri-Bio industry Technology Development Program, Export Promotion Technology Development Program (115067-02). Also this work was supported by Korea IPET through Advanced Production Technology Development Program (115102-03) and this work was supported by Rural Development Administration through Agriculture Science and Technology Development (Project No. PJ01408002).

References

  1. McMenamin AJ, Genersch E. Honey bee colony losses and associated viruses. Curr Opin Insect Sci 2015;8:121-129. https://doi.org/10.1016/j.cois.2015.01.015
  2. Genersch E, Yue C, Fries I, de Miranda JR. Detection of Deformed wing virus, a honey bee viral pathogen, in bumble bees (Bombus terrestris and Bombus pascuorum) with wing deformities. J Invertebr Pathol 2006;91:61-63. https://doi.org/10.1016/j.jip.2005.10.002
  3. Tehel A, Brown MJ, Paxton RJ. Impact of managed honey bee viruses on wild bees. Curr Opin Virol 2016;19:16-22. https://doi.org/10.1016/j.coviro.2016.06.006
  4. Reddy KE, Noh JH, Yoo MS, Kim YH, Kim NH, Doan HT, Ramya M, Jung SC, Van Quyen D, Kang SW. Molecular characterization and phylogenetic analysis of deformed wing viruses isolated from South Korea. Vet Microbiol 2013;167:272-279. https://doi.org/10.1016/j.vetmic.2013.08.018
  5. Tentcheva D, Gauthier L, Jouve S, Canabady-Rochelle L, Dainat B, Cousserans F, Colina ME, Ballb BV, Bergoin M. Polymerase chain reaction detection of deformed wing virus (DWV) in Apis mellifera and Varroa destructor. Apidologie (Celle) 2004;35:431-439. https://doi.org/10.1051/apido:2004021
  6. Chen YP, Higgins JA, Feldlaufer MF. Quantitative real-time reverse transcription-PCR analysis of deformed wing virus infection in the honeybee (Apis mellifera L.). Appl Environ Microbiol 2005;71:436-441. https://doi.org/10.1128/AEM.71.1.436-441.2005
  7. Yoo MS, Thi KC, Van Nguyen P, Han SH, Kwon SH, Yoon BS. Rapid detection of sacbrood virus in honeybee using ultra-rapid real-time polymerase chain reaction. J Virol Methods 2012;179:195-200. https://doi.org/10.1016/j.jviromet.2011.10.014
  8. Jamnikar Ciglenecki U, Toplak I. Development of a real-time RT-PCR assay with TaqMan probe for specific detection of acute bee paralysis virus. J Virol Methods 2012;184:63-68. https://doi.org/10.1016/j.jviromet.2012.05.010
  9. Haddad NJ, Noureddine A, Al-Shagour B, Loucif-Ayad W, El-Niweiri MA, Anaswah E, Hammour WA, El-Obeid D, Imad A, Shebl MA, Almaleky AS, Nasher A, Walid N, Bergigui MF, Yanez O, de Miranda JR. Distribution and variability of deformed wing virus of honeybees (Apis mellifera) in the Middle East and North Africa. Insect Sci 2017;24:103-113. https://doi.org/10.1111/1744-7917.12277
  10. Kukielka D, Esperon F, Higes M, Sanchez-Vizcaino JM. A sensitive one-step real-time RT-PCR method for detection of deformed wing virus and black queen cell virus in honeybee Apis mellifera. J Virol Methods 2008;147:275-281. https://doi.org/10.1016/j.jviromet.2007.09.008
  11. Lim HY. Development of novel rapid detection method for deformed wing virus (DWV) using ultra-fast high-performance PCR (UF-HP PCR). J Apic 2013;28:237-244.
  12. Ma M, Ma C, Li M, Wang S, Yang S, Wang S. Loop-mediated isothermal amplification for rapid detection of Chinese sacbrood virus. J Virol Methods 2011;176:115-119. https://doi.org/10.1016/j.jviromet.2011.05.028
  13. Yoo MS, Noh JH, Yoon BS, Reddy KE, Kweon CH, Jung SC, Kang SW. Reverse transcription loop-mediated isothermal amplification for sensitive and rapid detection of Korean sacbrood virus. J Virol Methods 2012;186:147-151. https://doi.org/10.1016/j.jviromet.2012.08.009
  14. Han SH, Lee DB, Lee DW, Kim EH, Yoon BS. Ultra-rapid real-time PCR for the detection of Paenibacillus larvae, the causative agent of American Foulbrood (AFB). J Invertebr Pathol 2008;99:8-13. https://doi.org/10.1016/j.jip.2008.04.010
  15. Lee DW, Kim EH, Yoo MS, Han SH. Ultra-rapid real-time PCR for the detection of human immunodeficiency virus (HIV). Korean J Microbiol 2007;43:91-99.
  16. Lim SJ, Kim JM, Lee CW, Yoon BS. Development of ultra-rapid multiplex PCR detection against 6 major pathogens in honeybee. J Apic 2017;32:27-39. https://doi.org/10.17519/apiculture.2017.04.32.1.27
  17. Lung O, Fisher M, Beeston A, Hughes KB, Clavijo A, Goolia M, Pasick J, Mauro W, Deregt D. Multiplex RT-PCR detection and microarray typing of vesicular disease viruses. J Virol Methods 2011;175:236-245. https://doi.org/10.1016/j.jviromet.2011.05.023
  18. Choe SE, Nguyen LT, Noh JH, Koh HB, Jean YH, Kweon CH, Kang SW. Prevalence and distribution of six bee viruses in Korean Apis cerana populations. J Invertebr Pathol 2012;109:330-333. https://doi.org/10.1016/j.jip.2012.01.003
  19. Tantillo G, Bottaro M, Di Pinto A, Martella V, Di Pinto P, Terio V. Virus infections of honeybees Apis Mellifera. Ital J Food Saf 2015;4:5364.
  20. Cai HY, Caswell JL, Prescott JF. Nonculture molecular techniques for diagnosis of bacterial disease in animals: a diagnostic laboratory perspective. Vet Pathol 2014;51:341-350. https://doi.org/10.1177/0300985813511132
  21. Wang JH, Lee DB, Ku SJ, Peak MC, Min SH, Lim SJ, Lee CW, Yoon BS. Development of a detection method against 11 major pathogens of honey bee using amplification of multiplex PCR and specific DNA-chip. J Apic 2016;31:133-146.