DOI QR코드

DOI QR Code

Comparison of Real-Time PCR and Culture Methods for Detection of Campylobacter jejuni in Various Foods

다양한 식품에서 Campylobacter jejuni 검출을 위한 real-time PCR과 배지배양법의 비교검증

  • Received : 2010.10.14
  • Accepted : 2010.11.22
  • Published : 2011.02.28

Abstract

In this study, performances of culture methods using two selective media and real-time PCR were evaluated for detection of Campylobacter jejuni (C. jejuni) in various food samples. Sausage, ground beef, and radish sprouts inoculated with C. jejuni were enriched in Hunt broth and then streaked onto modified cefoperazone charcoal deoxycholate agar and Preston agar, followed by incubation under microaerobic conditions. The enriched Hunt broth (1 mL) was used in real-time PCR assay. No statistical differences were observed in sensitivity among the two selective media and real-time PCR for sausage and ground beef. However, the number of positives by real-time PCR in radish sprouts was much higher than the two selective media (p<0.05). It appears that real-time PCR could be used as an effective screening tool to detect C. jejuni, particularly in foods with a high number of background microflora such as fresh vegetables.

Keywords

Campylobacter jejuni;real-time PCR;culture method;selective media

References

  1. Park CE, Sanders GW. Occurrence of thermotolerant campylobacters in fresh vegetables sold at farmers' outdoor markets and supermarkets. Can. J. Microbiol. 38: 313-316 (1992) https://doi.org/10.1139/m92-052
  2. Frost JA, Gillespie IA, O'Brien SJ. Public health implications of Campylobacter outbreaks in England and Wales, 1995-9: Epidemiological and microbiological investigations. Epidemiol. Infect. 128: 111-118 (2002)
  3. Brandl MT, Haxo AF, Bates AH, Mandrell RE. Comparison of survival of Campylobacter jejuni in the phyllosphere with that in the rhizosphere of spinach and radish plants. Appl. Environ. Microb. 70:1182-1189 (2004) https://doi.org/10.1128/AEM.70.2.1182-1189.2004
  4. Shin SY, Kim KY, Park JH. Survival of Campylobacter jejuni under aerobic condition. Korean J. Food sci. Technol. 30: 916-923 (1998)
  5. Lee YD, Park JH. Selective detection of Campylobacter jejuni, C. coli, Arcobacter butzleri and Helicobacter pylori by polymerase chain reaction. Korean J. Food Sci. Technol, 34: 1134-1139 (2002)
  6. Karenlampi R, Hänninen ML. Survival of Campylobacter jejuni on various fresh produce. Int. J. Food Microbiol. 97: 187-195 (2004) https://doi.org/10.1016/j.ijfoodmicro.2004.04.019
  7. Sahin O, Kobalka P, Zhang Q. Detection and survival of Campylobacter in chicken eggs. J. Appl. Microbiol. 95: 1070-1079 (2003) https://doi.org/10.1046/j.1365-2672.2003.02083.x
  8. Belanger AE, Shryock TR. Macrolide-resistant Campylobacter: the meat of the matter. J. Antimicrob. Chemoth. 60: 715-723 (2007) https://doi.org/10.1093/jac/dkm300
  9. Korea Food and Drug Administration. Food code. Available from http://safefood.kfda.go.kr/RS/food_menu.jsp, Accessed Dec. 10, 2009.
  10. Corry JE, Post DE, Colin P, Laisney MJ. Culture media for the isolation of campylobacters. Int. J. Food Microbiol. 26: 43-76 (1995) https://doi.org/10.1016/0168-1605(95)00044-K
  11. Bolton FJ, Hutchinson DN, Coates D. Blood-free selective medium for isolation of Campylobacter jejuni from feces. J. Clin. Microbiol. 19: 169-171 (1984)
  12. Moran L, Kelly C, Madden RH. Factors affecting the recovery of Campylobacter spp. from retail packs of raw, fresh chicken using ISO 10272-1:2006. Lett. Appl. Microbiol. 48: 628-632 (2009) https://doi.org/10.1111/j.1472-765X.2009.02581.x
  13. Gun-Munro J, Rennie RP, Thornley JH, Richardson HL, Hodge D, Lynch J. Laboratory and clinical evaluation of isolation media for Campylobacter jejuni. J. Clin. Microbiol. 25: 2274-2277 (1987)
  14. Peterz M. Comparison of Preston agar and a blood-free selective medium for detection of Campylobacter jejuni in food. J. Assoc. Off. Anal. Chem. 74: 651-654 (1991)
  15. Karmali MA, Simor AE, Roscoe M, Fleming PC, Smith SS, Lane J. Evaluation of a blood-free, charcoal-based, selective medium for the isolation of Campylobacter organisms from feces. J. Clin. Microbiol. 23: 456-459 (1986)
  16. Line JE, Stern NJ, Lattuada CP, Benson ST. Comparison of methods for recovery and enumeration of Campylobacter from freshly processed broilers. J. Food Protect. 64: 982-986 (2001)
  17. Bolton FJ, Sails AD, Fox AJ, Wareing DR, Greenway DL. Detection of Campylobacter jejuni and Campylobacter coli in foods by enrichment culture and polymerase chain reaction enzyme-linked immunosorbent assay. J. Food Protect. 65: 760-767 (2002)
  18. Josefsen MH, Lubeck PS, Aalbaek B, Hoorfar J. Preston and Park-Sanders protocols adapted for semi-quantitative isolation of thermotolerant Campylobacter from chicken rinse. Int. J. Food Microbiol. 80: 177-183 (2003) https://doi.org/10.1016/S0168-1605(02)00151-4
  19. Sachse K. Specificity and performance of diagnostic PCR assays. Methods Mol. Biol. 216: 3-29 (2003)
  20. Yang C, Jiang Y, Huang K, Zhu C, Yin Y. Application of realtime PCR for quantitative detection of Campylobacter jejuni in poultry, milk, and environmental water. FEMS Immunol. Med. Mic. 38: 265-271 (2003) https://doi.org/10.1016/S0928-8244(03)00168-8
  21. Hyeon JY, Hwang IG, Kawk HS, Park JS, Heo S, Choi IS, Park CK, Seo KH. Evaluation of an automated ELISA and real-time PCR by comparing with a conventional culture method for the detection of Salmonella spp. in steamed pork and raw broccoli sprouts. Korean J. Food Sci. Anim. Resour. 29: 506-512 (2009) https://doi.org/10.5851/kosfa.2009.29.4.506
  22. U.S. Food and Drug Administration. Bacteriological Analytical Manual, Chapter7 Campylobacter. Available at: http://www.fda.gov/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/UCM072616. Accessed June 3 2010.
  23. Lee JH, Song KY, Hyeon JY, Hwang IG, Kwak HS, Han JA, Chung YH, Seo KH. Comparison of standard culture method and real-time PCR assay for detection of Staphylococcus aureus in processed and unprocessed foods. Korean J. Food Sci. Anim. Resour. 30: 410-418 (2010) https://doi.org/10.5851/kosfa.2010.30.3.410
  24. Han SR, Hyeon JY, Kim HY, Park JS, Heo S, Shin HC, Seo KH. Evaluation of conventional culture methods and validation of immunoassays for rapid detection of Listeria monocytogenes in dairy and processed foods. Korean J. Food Sci. Anim. Resour. 28: 616-622 (2008) https://doi.org/10.5851/kosfa.2008.28.5.616
  25. Seo KH, Brackett RE. Rapid, specific detection of Enterobacter sakazakii in infant formula using a real-time PCR assay. J. Food Protect. 68: 59-63 (2005)
  26. Best EL, Powell EJ, Swift C, Grant KA, Frost JA. Applicability of a rapid duplex real-time PCR assay for speciation of Campylobacter jejuni and Campylobacter coli directly from culture plates. FEMS Microbiol. Lett. 229: 237-241 (2003) https://doi.org/10.1016/S0378-1097(03)00845-0
  27. Chon JH, Hyeon JY, Hwang IG, Kwak HS, Han JA, Chung YH, Song KW, Seo KH. Comparison of standard culture method and real-time PCR for detection of Vibrio parahaemolyticus in seafoods and vegetables. Korean J. Food Sci. Technol. 42: 355-360 (2010)
  28. Evans MR, Ribeiro CD, Salmon RL. Hazards of healthy living: bottled water and salad vegetables as risk factors for Campylobacter infection. Emerg. Infect. Dis. 9: 1219-1225 (2003) https://doi.org/10.3201/eid0910.020823
  29. Hussain I, Shahid Mahmood M, Akhtar M, Khan A. Prevalence of Campylobacter species in meat, milk and other food commodities in Pakistan. Food Microbiol. 24: 219-222 (2007) https://doi.org/10.1016/j.fm.2006.06.001
  30. Luber P, Brynestad S, Topsch D. Quantification of Campylobacter species cross-contamination during handling of contaminated fresh chicken parts in chicken in kitchens. Appl. Environ. Microb. 72: 66-70 (2006) https://doi.org/10.1128/AEM.72.1.66-70.2006
  31. Malorny B, Paccassoni E, Fach P, Bunge C, Martin A, Helmuth R. Diagnostic real-time PCR for detection of Salmonella in food. Appl. Environ. Microb. 70: 7064-7052 (2004)
  32. Cai T, Jiang L, Yang C, Huang K. Application of real-time PCR for quantitative detection of Vibrio parahaemolyticus from seafood in eastern China. FEMS Immunol. Med. Mic. 46: 180-186 (2006) https://doi.org/10.1111/j.1574-695X.2005.00016.x

Cited by

  1. Comparative Evaluation of Real-Time PCR and Conventional Culture Method Using Two Selective Agars for the Detection of Cronobacter spp. in Powdered Infant Formula and Dried Pumpkin vol.31, pp.6, 2016, https://doi.org/10.13103/JFHS.2016.31.6.439
  2. Comparison of Conventional Culture Method and Real-time PCR for Detection of Yersinia enterocolitica in Sausage and Vegetable Salad vol.45, pp.1, 2013, https://doi.org/10.9721/KJFST.2013.45.1.133
  3. Improvement of Enterobacteriaceae enrichment broth by supplementation with sodium citrate for detection of Cronobacter sakazakii using real-time PCR vol.25, pp.4, 2016, https://doi.org/10.1007/s10068-016-0191-7

Acknowledgement

Supported by : 농촌진흥청