KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
권호 표지
DOI QR코드

DOI QR Code

The Early Detection of the Spore Using Sonication and Fluorescent Dye in the Field

현장에서 초음파 파쇄와 형광시약을 이용한 포자의 조기 탐지

  • Received : 2011.04.19
  • Accepted : 2011.08.08
  • Published : 2011.08.30
Download PDF
⟨ Previous Next ⟩

Abstract

This study was carried out to establish the optimum condition of cell disruption with a sonicator for the detection of the spore, Bacillus anthracis ${\Delta}$-sterne for the purpose of developing automatic fluorometer. The efficiency of sonication on the ${\Delta}$-sterne spore disruption was very weak. The ${\Delta}$-sterne spore with zirconia bead showed greater disruption than the ${\Delta}$-sterne spore alone when sonificated. The volumn of the zirconia bead added in the spore solution has little effect on the disruption efficiency. The detection limit of the ${\Delta}$-sterne spore with zirconia bead and the ${\Delta}$-sterne spore alone was $10^6$ CFU/mL and $5{\times}10^7$ CFU/mL respectively, when sample was sonicated for 20 seconds with a sonicator probe of 13 mm diameter.

Keywords

References

  1. Reisa, J. (1999) Chemical and Biological Terrorism. National Academy Press, Washington, D. C. USA.
  2. Turnbull, P. C., R. Bohm, O. Cosivi, M. Doganay, M. E. Hugh- Jones, D. D. Joshi, and M. K. Lalitha (1998) Guidelines for the Surveillance and Control of Anthrax in Human and Animals, WHO/EMC/ZDI 3rd ed. pp. 12-18. Switzerland.
  3. Chandler, D. P., J. Brown, C. J., Bruckner-Lea, L. Olson, G. J. Posakony, J. R. Stults, N. B. Valentine, and L. J. Bond (2001) Continuous spore disruption using radially focused, highfrequency ultrasound. Anal. Chem. 73: 3784-3789. https://doi.org/10.1021/ac010264j
  4. Marentis, T. C., B. Kusler, G. G. Yaralioglu, S. Liu, E. O. Hæggström, and B. T. Khuri-Yakub (2005) Microfluidic sonicator for real-time disruption of eukaryotic cells and bacterial spores for DNA analysis. Ultrasound in Med. & Biol. 31: 1265-1277. https://doi.org/10.1016/j.ultrasmedbio.2005.05.005
  5. Belgrader, P., D. Hansford, G. T. A. Kovacs, K. Venkateswaran, R. Mariella, Jr. F. Milanovich, S. Nasarabadi, M. Okuzumi, F. Pourahmadi, and M. A. Northrup (1999) A minisonicator to rapidly disrupt bacterial spores for DNA analysis. Anal. Chem. 71: 4232-4236. https://doi.org/10.1021/ac990347o
  6. Taylor, M. T., P. Belgrader, B. J. Furman, F. Pourahmadi, G. T. A. Kovacs, and M. A. Northrup (2001) Lysing bacterial spores by sonification through a flexible interface in a microfluidic system. Anal. Chem. 73: 492-496. https://doi.org/10.1021/ac000779v
  7. Kuske, C. R., K. L. Banton, D. L. Adorada, P. C. Stark, K. K. Hill, and P. J. Jackson (1998) Small-scale DNA sample preparation method for field PCR detection of microbial cells and spores in soil. Appl. Environ. Microbiol. 64: 2463-2472.
  8. More, M. I., J. B. Herrick, M. C. Silva, W. C. Ghiorse, and E. L. Madsen (1994) Quantitative cell lysis of indigenous microorganisms and rapid extraction of microbial DNA from sediment. Appl. Environ. Microbiol. 60: 1572-1580.
  9. Leighton, T. J. and R. H. Doi (1971) The stability of messenger ribonucleic acid during sporulation in Bacillus subtilis. J. Biol. Chem. 246: 3189-3195.