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Development of Nested Ultra-Rapid PCR Method for the Accurate Detection of Acute Bee Paralysis Virus (ABPV)

Acute Bee Paralysis Virus (ABPV)의 정확한 검출을 위한 Nested 초고속 PCR법 개발

  • Kim, MoonJung (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Kim, JungMin (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Kim, ByeongHee (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Kim, SoMin (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Truong, A Tai (Department of Life Science, College of Fusion Science, Kyonggi University) ;
  • Yoon, ByoungSu (Department of Life Science, College of Fusion Science, Kyonggi University)
  • 김문정 (경기대학교 융합과학대학 바이오융합학부 생명과학전공) ;
  • 김정민 (경기대학교 융합과학대학 바이오융합학부 생명과학전공) ;
  • 김병희 (경기대학교 융합과학대학 바이오융합학부 생명과학전공) ;
  • 김소민 (경기대학교 융합과학대학 바이오융합학부 생명과학전공) ;
  • ;
  • 윤병수 (경기대학교 융합과학대학 바이오융합학부 생명과학전공)
  • Received : 2018.09.13
  • Accepted : 2018.09.27
  • Published : 2018.09.30

Abstract

Acute bee paralysis virus (ABPV) is an infectious disease causing paralysis in healthy honeybee colonies. Because ABPV is highly homologous with relative viruses, it is difficult to distinguish ABPV from especially IAPV or KBV by molecular diagnosis. In this study, for accurate and specific detection of ABPV, a secondary nested PCR detection method following the primary detection PCR against ABPV was developed. It could be able to detect from less than 100 molecules of ABPV. The detection time was also minimized from RNA extraction to PCR. Using ultra-rapid ABPV-specific detection and nested PCR, the presence of ABPV was detected from infected bee samples within 50 minutes. It might be expected that this kind of detection would be helpful for accurate detection and monitoring of ABPV in environment.

Acknowledgement

Supported by : 농촌진흥청, 농림수산식품기술기획평가원, 중소기업청

References

  1. 민상현, 왕지희, 임수진, 이칠우, 윤병수. 2016. 꿀벌 병원성 바이러스의 정량적 검출을 위한 정량 실시간 재조합 효소-중합효소 증폭법의 개발. Journal of Apiculture 31(2): 147-156.
  2. 왕지희, 이도부, 구수진, 백문철, 민상현, 임수진, 이칠우, 윤병수. 2016. 다중PCR증폭과 특이 DNA-chip을 활용한 꿀벌 주요 11종 병원체의 검출법 개발. Journal of Apiculture 31(2): 133-146.
  3. 유미선, 이동우, 김일욱, 김동수, 권순환, 임윤규, 윤병수. 2007. 국내 양봉장에서 Kashmir bee virus(KBV)의 검출과 이의 분자생물학적 고찰. Journal of Apiculture 22(1): 33-42.
  4. 이중구, 이보람, 윤병수. 2012. Loop-mediated isothermal amplification(LAMP) 법을 이용한 Kakugo virus의 새로운 검출법 개발. Journal of Apiculture 27(4): 267-274.
  5. 이혜민, 이도부, 한상훈, 이명렬, 임윤규, 윤병수. 2005. 한국산 꿀벌에서 Deformed wing virus의 발견과 real-time PCR 검출법의 원리. Journal of apiculture 20(2): 85-94.
  6. 최용수, 이명렬, 이만영. 2007. 국내 꿀벌에서의 Israle acute paralysis virus(IAPV) 진단. Journal of apiculture 22(2): 159-165.
  7. Bailey, L., Gilbbs A. J., Woods R.D. 1963. Two viruses from adult honey bees (Apis mellifera Linnaeus). Journal of virology 21(3): 390-395.
  8. Bakony, Tamas, Farkas Robert, Szendroi Amdrea, Dobos-Kovacs Mihaly and Rusvai Mikols. 2002. Detection of acute bee paralysis virus by RT-PCR in honey bee and Varroa destructor field samples: rapid screening of representative Hungarian apiaries. Apidologie 33: 63-74.
  9. Carpana, E., Vecchi M. A., Lavazza A., Bassi S.,Dottori M. and Ritter W. 1991. Prevalence of acute paralysis virus (APV) and other viral infections in honeybees in Italy. Proc. Int. Symp. on Bee Pathology, Ghent, Belgium, pp. 155-165.
  10. Chen, Yan Ping and Siede Reinhold. Honey bee virus. 2007. Advances in virus research 70: 33-80.
  11. de Miranda, Joachim R., Cordoni Guido and Budge Giles. 2010. The acute bee paralysis virus-Kashmir bee virus-Israeli acute paralysis virus complex. Journal of Invertebrate Pathology 103: S30-S47.
  12. de Miranda, Joachim R., Dainat Bejamin, Locke Barbara, Cordoni Guido, Berthoud Helene, Gauthier Laurent, Neumann Peter, Budge Giles E., Ball Brenda V. and Stoltz Don B. 2010. Genetic characterization of slow bee paralysis virus of the honeybee (Apis mellifera L.). Journal of General Virology 91: 2524-2530.
  13. Faucon, J. P., Vitu C., Russo P. and Vignoni M. 1992. Diagnosis of acute paralysis: application to the epidemiology of honey bee viral diseases in France in 1990, Apidologie 23: 139-146.
  14. Hung, A.C.F. and Shimanuki H. 1999. A scientific note on the detection of Kashmir bee virus in individual honeybees and Varroa jacobsoni mites, Apidologie 30: 353-354.
  15. McMahon, Dino P., Wilfert Lena, Paxton Robert J. and Brown Mark J. F. 2018. Emerging viruses in bees: From molecules to ecology. Advances in virus research 101: 251-291.