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Immunocapture RT-PCR for Detection of Seed-borne Viruses on Cucurbitaceae Crops

Immunocapture RT-PCR을 이용한 박과작물 종자전염 바이러스의 검출

  • Received : 2010.03.16
  • Accepted : 2010.07.24
  • Published : 2010.08.01

Abstract

Immunocapture reverse transcription polymerase chain reaction (IC-RT-PCR) was applied to the detection of Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), and Zucchini green mottle mosaic virus (ZGMMV) on Cucurbitaceae crops. These seed-borne tobamoviruses were accurately detected from the infected leaves and seeds by IC-RT-PCR. This method was estimated to be about 100 times more sensitive than ELISA, and also it allowed the direct confirmation of ELISA results by using the captured antigens from a completed ELISA microwell. This convenient and reliable method could be used routinely for large-scale field surveys or seed tests of Cucurbitaceae crops.

박과작물에 발생하는 종자전염 바이러스 3종(CGMMV, KGMMV, ZGMMV)에 대한 검출을 위해 IC-RT-PCR의 적용 가능성을 시험하였다. IC-RT-PCR을 이용하여 감염 종자와 잎으로 부터 바이러스의 특이적인 검출이 가능하였으며, 검출 민감도는 ELISA 보다 100배 이상 높았다. 또한 반응을 마친 ELISA 마이크로플레이트에 남아있는 항원을 IC-RT-PCR의 template로 사용할 경우 ELISA 결과를 곧바로 확인할 수 있었다. 따라서 IC-RT-PCR에 의한 본 검정방법은 박과작물의 대규모 포장검사 및 종자검사에 편리하게 사용될 수 있을 것으로 기대된다.

Keywords

References

  1. 조점덕, 김정수, 이신호, 정봉남. 2007. 박과 작물 종자전염 바이러스 3종(CGMMV, ZGMMV, KGMMV)의 간편한 동시진단 VC/RT-PCR 유전자 진단. 식물병연구 13: 82-87. https://doi.org/10.5423/RPD.2007.13.2.082
  2. Harness, A. M., Johnson, J. C., Kulemeka, B. P., Sutula, C. L. and Bandla, M. D. 2003. Immunocapture RT-PCR as a confirmation tool for ELISA. Phytopathology 92: S34.
  3. 김정수, 이수헌, 최홍수, 최국선, 조점덕, 정봉남. 2008. 2007년 우리나라 주요 작물 바이러스병 발생 상황. 식물병연구 14: 1-9. https://doi.org/10.5423/RPD.2008.14.1.001
  4. 고숙주, 이용환, 차광홍, 박진우, 이수헌, 양광열. 2004. 전남지역의 수박에 발생하는 바이러스 병 발생 실태. 식물병연구 10: 39-43. https://doi.org/10.5423/RPD.2004.10.1.039
  5. Ko, S. J., Lee, Y. H., Cho, M. S., Park, J. W., Choi, H. S., Lim, G. C. and Kim, K. H. 2007. The incidence of virus disease on Melon in Jeonnam province during 2000-2002. Plant Pathol. J. 23: 215-218. https://doi.org/10.5423/PPJ.2007.23.3.215
  6. Lee, S. H., Kim, S. M., Kim, W. C. and Lee, K. W. 2003. Multiplex reverse transcription polymerase chain reaction assay for simultaneous detection of five cucurbit-infecting viruses. Plant Pathol. J. 19: 342.
  7. 이숙경, 송완엽, 김형무. 2004. RT-PCR에 의한 박 종자의 오이녹반모자이크바이러스 검정. 식물병연구 10: 53-57. https://doi.org/10.5423/RPD.2004.10.1.053
  8. Olmos, A., Dasi, M. A., Candresse, T. and Cambra, M. 1996. Print-capture PCR: a simple and highly sensitive method for the detection of Plum pox virus (PPV) in plant tissues. Nucleic Acids Research 24: 2192-2194. https://doi.org/10.1093/nar/24.11.2192
  9. Rampersad, S. N. and Umaharan, P. 2003. Detection of Begomoviruses in clarified plant extracts: A comparison of standard, direct-binding, and immunocapture PCR techniques. Phytophathology 93: 1153-1157. https://doi.org/10.1094/PHYTO.2003.93.9.1153
  10. Rowhani, A., Maningas, M. A., Lile, L. S., Daubert, S. D. and Golino, D. A. 1995. Development of a detection system for viruses of woody plants based on PCR analysis of immobilized virions. Phytopathology 85: 347-352. https://doi.org/10.1094/Phyto-85-347
  11. Wetzel, T., Candresse, T., Macquaire, G., Ravelonadro, M. and Dunez, J. 1992. A highly sensitive immunocapture polymerase chain reaction method for plum pox potyvirus detection. J. Virol. Methods 39: 27-37. https://doi.org/10.1016/0166-0934(92)90122-T

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