한국식품위생안전성학회지 (Journal of Food Hygiene and Safety)

  • 제18권4호
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  • Pages.224-228
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  • 2003
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  • 1229-1153(pISSN)
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  • 2465-9223(eISSN)
한국식품위생안전성학회 (The Korean Society of Food Hygiene and Safety)
권호 표지

Salmonella spp.의 RAPD Typing을 위한 Primer의 분리력 비교

Primers for Typing Salmonella spp. using Random Amplified Polymorphic DNA (RAPD) Analysis

  • 발행 : 2003.12.01
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초록

RAPD 분석은 한 개의 primer를 이용하여 임의의 DNA조각을 증폭하는 것이다. 이때 만들어지는 여러 형태의 DNA band유형을 이용하여 살모넬라를 분류할 수 있다. 살모넬라를 효과적으로 RAPD typing할 수 있는 primer들을 엄선하기 위하여 살모넬라 표준군주 16종을 대상으로 총 20가지 primer들의 RAPD 분리력을 비교하여 보았다. 결과는 재현성이 높았으며 이중 primer A, OPG04, OPG10, OPL-03을 16가지 균 모두를 다른 유형으로 분리하였고 primer OPB-17, OPB-6, OPG08, OPL-02는 15가지 유형으로 분리하였다. 이들은 discrimination index, band의 숫자, band scoring의 난이도 등을 고려해 볼 때 나머지 primer 들에 비해 우수한 분리력을 보였다. 이들 primer들은 장창 살모넬라의 RAPD typing에 이용될 수 있을 것으로 보이며, 현재는 이들을 이용하여 돈육 공장의 오염원 구명을 위한 연구를 수행 중이다.

Random amplified polymorphic DNA (RAPD) analysis is based on the amplification of random DNA segment using a single arbitratrary primer. For typing Salmonella spp., polymorphic DNA patterns identified by this method can be used. To select the primers for RAPD typing Salmonella spp., the performances of 20 primers were compared by analyzing 16 Salmonella spp. reference strains. Reproducible electrophoresis patterns were obtained. Among the 20 primers tested, 4 primers (A, OPG04, OPG10, OPL03) showed better differentiation than the others. At the time discrimination index, band clarity, band number and difficulty of band scoring were considered. These primers will be useful for typing Salmonella spp. in the future. Curretly, we are under investigation for the RAPD typing of contaminated Slmonella spp. for the risk analysis of pork processing plant using the primers.

키워드

참고문헌

  1. Olsen, J.E., Aabo, S., Hill, W., Notermans, S., Wernars, K., Granum, P.E., Popovic, T, Rasmussen, H.N. and Olsvik, O: Probes and polymerase chain reaction for the detection of food-borne bacterial pathogens. International J. Food Microbiol., 28, 1-78 (1995) https://doi.org/10.1016/0168-1605(94)00159-4
  2. Carrington, M. and Hoelzel, A.R.: Molecular Epidemiology. Oxford University Press, pp 41-43 (2001)
  3. Kerouanton, A, Brisabois, A., Denoyer, E., Dilasser, F., Grout, J., Salvat, G. and Picard, B.: Comparison of five typing methods for the epidemiological study of Listeria monocytogenes. International J. Food Microbiol., 43, 61-71 (1998) https://doi.org/10.1016/S0168-1605(98)00098-1
  4. Soto, S.M., Guerra, B., Gonzalez-Hevia, M.A. and Mendoza, M.C.: Potential of three-way randomly amplified polymorphic DNA analysis as a typing method for twelve Salmonella serotypes. Appl. Environ. Microbiol.. 65, 4830-4836 (1999)
  5. Mare, L., Dicks, L.M.T. and van der Walt, M.L.: Characterization of south African isolates of Salmonella enteritidis by phage typing, numerical analysis of RAPD-PCR banding patterns and plasmid profiles. International, J. Food Microbiol., 64, 237-245 (2001) https://doi.org/10.1016/S0168-1605(00)00448-7
  6. Millemann, Y., Lesage-Descauses, M.-C., Lafont, J.-P. and Chaslus-Dancla, E.: Comparison of randomly amplified polymorphic DNA analysis and enterobacterial repetitive intergenic consensus-PCR for epidemiological studies of Salmonella. FEMS Immunol. & Microbiol., 14, 129-134 (1996) https://doi.org/10.1111/j.1574-695X.1996.tb00279.x
  7. Martin, M.C., Gonzalez-Hevia, M.A., Moro, I. and Mendoza, M.C.: Genetic typing methods applied to the differentiation of clonal lines among Salmonella enterica serogroup G strains causing human salmonellosis. FEMS Immunol. & Microbiol., 19, 215-221 (1997) https://doi.org/10.1016/S0928-8244(97)00086-2
  8. Landeras, E., Gonzalez-Hevia, M.A. and Mendoza, M.C.: Molecular epidemiology of Salmonella serotype enteritidis. Relationships between food, water and pathogenic strains. International J. Food Microbiol., 43, 81-90 (1998) https://doi.org/10.1016/S0168-1605(98)00092-0
  9. Laconcha, I., Lopez-Molina, N., Rementeria, A., Audicana, A., Perales, I. and Garaizar, J.: Phage typing combined with pulsed-field gel electrophoresis and random amplified polymorphic DNA increases discrimination in the epidemiological analysis of Salmonella enteritidis strains. International J. Food Microbiol., 40, 27-34 (1998) https://doi.org/10.1016/S0168-1605(98)00007-5
  10. Tsen, H., Hu, H.H., Lin, J.S., Huang, C.H. and Wang, T.K.: Analysis of the Salmonella typhimurium isolates from foodpoisoning cases by molecular subtyping methods. Food Microbiol., 17, 143-152 (2000) https://doi.org/10.1006/fmic.1999.0284
  11. Choi, W.S. and Hong, C.-H.:Rapid enumeration of Listeria monocytogenes in milk using competitive PCR. International J. Food Microbial., 84, 79-85 (2003) https://doi.org/10.1016/S0168-1605(02)00401-4
  12. Hunter, P.R. and Gaston, M.A.: Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity. J. Clin. Microbiol., 26, 2465-2466 (1998)
  13. Van de Peer, Y., and De Wachter, R.: TREECON for Windows: a software package for the construction and drawing of evolutionary trees for the Microsoft Windows environment. Comput. Applic. Biosci., 10, 569-570 (1994)
  14. Link, W., Dixens, C., Singh, M., Schwall, M. and Melchinger, A.E.: Genetic diversity in European and Mediterranean faba bean germ plasm revealed by RAPD markers. Theor. Appl. Genet., 90, 27-32 (1995)
  15. Lin, A.W., Usera, M.A., Barret, T.J. and Goldsby, R.A.: Application of random amplified polymorphic DNA analysis to differentiate strains of Salmonella enteritidis. J. Clin. Microbiol., 34, 870-876 (1996)
  16. Williams J.G.K., Kubelin, A.R., Livak, K.J., Rafalski, J.A. and Tingey, S.V.: DNA polymorphism amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res., 18, 6531-6535 (1990) https://doi.org/10.1093/nar/18.22.6531
  17. Miyata, M., Aoki, T., Inglis, V., Yoshida, T. and Endo, M.: RAPD analysis of Aeromonas salmonicida and Aeromonas hydrophila. J. Appl. Bacteriol., 79, 181-185 (1995) https://doi.org/10.1111/j.1365-2672.1995.tb00933.x
  18. Chansiripornchai, N., Ramasoota, P., Bangtrakulnonth, A, Sasipreeyajan, J. and Svenson, S.B.: Application of randomly amplified polymorphic DNA (RAPD) analysis for typing avian Salmonella enterica subsp. enterica. FEMS Immunol. Med. Microbiol., 29, 221-225 (2000) https://doi.org/10.1111/j.1574-695X.2000.tb01526.x
  19. Martinez, I., Rorvik, L.-M., Brox, V., Lassen, J., Seppola, M., Gram, L. and Fonnesbech-Vogel, B.: Genetic variability among isolates of Listeria monocytogenes from food products, clinical samples and processing environments, estimated by RAPD typing. International J. Food Microbiol., 84, 285-297 (2003) https://doi.org/10.1016/S0168-1605(02)00423-3