Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
권호 표지
DOI QR코드

DOI QR Code

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

조제분유와 건조호박에서 Cronobacter spp. 검출을 위한 두 가지 선택배지와 Real-time PCR의 비교검증

  • Kim, Hong-Seok (Center for One Health, College of Veterinary Medicine, Konkuk University) ;
  • Shin, Minjung (Center for One Health, College of Veterinary Medicine, Konkuk University) ;
  • Chon, Jung-Whan (Center for One Health, College of Veterinary Medicine, Konkuk University) ;
  • Lim, Jong-Soo (Center for One Health, College of Veterinary Medicine, Konkuk University) ;
  • Kim, Young-Ji (Center for One Health, College of Veterinary Medicine, Konkuk University) ;
  • Kim, Dong-Hyeon (Center for One Health, College of Veterinary Medicine, Konkuk University) ;
  • Chang, Ho-Seok (Center for One Health, College of Veterinary Medicine, Konkuk University) ;
  • Kim, Hyunsook (Department of Food & Nutrition, College of Human Ecology, Hanyang University) ;
  • Om, Ae-Son (Department of Food & Nutrition, College of Human Ecology, Hanyang University) ;
  • Oh, Deog-Hwan (Department of Food Science and Biotechnology, School of Bioconvergence Science and Technology, Kangwon National University) ;
  • Song, Kwang-Young (Center for One Health, College of Veterinary Medicine, Konkuk University) ;
  • Seo, Kun-Ho (Center for One Health, College of Veterinary Medicine, Konkuk University)
  • 김홍석 (건국대학교 수의과대학 식품안전건강연구소) ;
  • 신민정 (건국대학교 수의과대학 식품안전건강연구소) ;
  • 천정환 (건국대학교 수의과대학 식품안전건강연구소) ;
  • 임종수 (건국대학교 수의과대학 식품안전건강연구소) ;
  • 김영지 (건국대학교 수의과대학 식품안전건강연구소) ;
  • 김동현 (건국대학교 수의과대학 식품안전건강연구소) ;
  • 장호석 (건국대학교 수의과대학 식품안전건강연구소) ;
  • 김현숙 (한양대학교 생활과학대학 식품영양학과) ;
  • 엄애선 (한양대학교 생활과학대학 식품영양학과) ;
  • 오덕환 (강원대학교 농업생명과학대학 식품생명공학과) ;
  • 송광영 (건국대학교 수의과대학 식품안전건강연구소) ;
  • 서건호 (건국대학교 수의과대학 식품안전건강연구소)
  • Received : 2016.07.22
  • Accepted : 2016.10.25
  • Published : 2016.12.30
Download PDF
⟨ Previous Next ⟩

Abstract

In the present study, the performance of culture methods using two selective agars and real-time PCR were compared for selective isolation of Cronobacter in powdered infant formula and dried pumpkin. Two food samples were spiked with the pathogen and then preenriched in distilled water. A small portion of preenrichment (10 mL) was incubated in Enterobacteriaceae enrichment both, followed by inoculation onto Druggan-Forsythe-Iversen agar (DFI agar) and Cronobacter sakazakii chromogenic plating agar (R&F agar). The preenrichment and enrichment (1 mL each) was used in real-time PCR assay. In powdered infant formula (PIF), no statistical difference was observed between both culture methods and real-time PCR with preenrichemt (p > 0.05). However, the number of positives obtained by R&F agar and real-time PCR was much higher than that of culture method using DFI agar in dried pumpkin (p < 0.05). In particular, R&F agar yielded a significantly greater selectivity than DFI agar in dried pumpkin (p < 0.05). Real-time PCR and R&F agar, which are currently recommended by US FDA, could be used as an alternative detection tools for the isolation of Cronobacter in PIF and ingredient of child foods such as dried pumpkin that has high number of competing natural microflora.

두 종류의 Cronobacter 선택배지(DFI agar, R&F agar)의 분유 및 건조호박 내 Cronobacter의 선택 분리능을 real-time PCR법과 함께 비교하였다. 분유에서의 Cronobacter 검출률은 세 가지 방법에서 유의적인 차이를 나타내지 않았으나(p < 0.05), 건조호박의 경우 R&F배지와 real-time PCR법이 DFI에서보다 유의적으로 높은 검출률을 보였다(p < 0.05). 배지 간 선택성에 있어서도, R&F 선택배지는 건조호박에서 DFI에 비해 유의적으로 높은 선택성을 나타냈다(p < 0.05). Real-time PCR 및 R&F배지의 사용은 분유뿐만 아니라, 건조 호박 등의 높은 경쟁세균총을 갖는 영유아식의 원료로 사용될 수 있는 식품군에서도 Cronobacter를 효과적으로 검출할 수 있는 방법으로 사료된다.

Keywords

References

  1. Burdette JH, Santos C.: Enterobacter sakazakii brain abscess in the neonate: the importance of neuroradiologic imaging. Pediatr. Radiol., 30(1), 33-34 (2000). https://doi.org/10.1007/s002470050009
  2. Gallagher PG, Ball WS.: Cerebral infarctions due to CNS infection with Enterobacter sakazakii. Pediatr. Radiol., 21(2), 135-136 (1991). https://doi.org/10.1007/BF02015629
  3. Liu Y, Cai X, Zhang X, Gao Q, Yang X, Zheng Z, Luo M, Huang X.: Real time PCR using TaqMan and SYBR Green for detection of Enterobacter sakazakii in infant formula. J. Microbiol. Meth., 65(1), 21-31 (2006). https://doi.org/10.1016/j.mimet.2005.06.007
  4. Lai KK.: Enterobacter sakazakii infections among neonates, infants, children, and adults. Case reports and a review of the literature. Medicine, 80(2), 113-122 (2001). https://doi.org/10.1097/00005792-200103000-00004
  5. Kim K, Jang SS, Kim SK, Park JH, Heu S, Ryu S.: Prevalence and genetic diversity of Enterobacter sakazakii in ingredients of infant foods. Int. J. Food Microbiol., 122(1), 196-203 (2008). https://doi.org/10.1016/j.ijfoodmicro.2007.11.072
  6. US Food and Drug Administration. Bacteriological Analytical Manual, Chapter 29, Cronobacter (2012) http://www.fda.gov/Food/FoodScienceResearch/LaboratoryMethods/ucm2-89378.htm (accessed December 13, 2016)
  7. Korea Food and Drug Administration. Korea Food Code, Chapter 5-29, Ready-to-Eat and Convenience Foods (2013).
  8. Korea Food and Drug Administration (2007). http://www.mfds.go.kr/index.do?seq=1151&mid=675 (accessed December 13, 2016).
  9. Lee EJ, Kim SG, Yoo SR, Oh SS, Hwang IG, Kwon GS, Park JH.: Microbial Contamination by Bacillus cereus, Clostridium perfringens, and Enterobacter sakazakii in Sunsik. Food Sci. Biotechnol., 16(6), 948-953 (2007).
  10. Chon JW, Song KY, Kim SY, Hyeon JY, Seo KH.: Isolation and Characterization of Cronobacter from Desiccated Foods in Korea. J. Food Sci., 77(7), 354-358 (2012). https://doi.org/10.1111/j.1750-3841.2012.02750.x
  11. Kandhai MC, Heuvelink, AE, Reij MW, Beumer, RR, Dijk R, van Tilburg JC, van Schothorst M, Gorris LG.: A study into the occurrence of Cronobacter spp. in The Netherlands between 2001 and 2005. Food Cont., 21(8), 1127-1136 (2010). https://doi.org/10.1016/j.foodcont.2010.01.007
  12. Lee EJ, Ryu TH, Park JH.: Tolerance of Korean Cronobacter spp. isolates to desiccation. Korean J. Food Sci. Technol., 41(6), 681-686 (2009).
  13. Song KY, Chon JW, Kim HS, Seo KH.: Current Cronobacter spp. Researches on Prevalence, Control, and Detection. Korean J. Microbiol., 48(4), 229-239 (2012). https://doi.org/10.7845/kjm.2012.051
  14. Chon JW, Song KY, Kim SY, Hyeon JY, KimYG, Seo KH.: Comparison of Real-Time PCR and Conventional Culture Method for Detection of Cronobacter spp. in Powdered Foods. Korean J. Microbiol., 47(1), 87-91 (2011).
  15. Chon JW, Hyeon JY, Hwang IG, Kwak HS, Han JA, Seo KH.: Comparison of Real-Time PCR and Culture Methods for Detection of Campylobacter jejuni in Various Foods. Korean J. Food Sci. Technol., 43(1), 119-123 (2011). https://doi.org/10.9721/KJFST.2011.43.1.119
  16. Kim YK, Chon JW, Lee JH, Kwak HS, Hwang IG, Seo KH.: Comparison of Conventional Culture Method and Real-time PCR for Detection of Yersinia enterocolitica in Sausage and Vegetable Salad. Korean J. Food Sci. Technol., 45(1), 133-136 (2013). https://doi.org/10.9721/KJFST.2013.45.1.133
  17. Hyeon JY, Hwang IG, Kwak HS, Park JS, Heo S, Seo KH.: Evaluation of an Automated ELISA (VIDAS(R)) 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(4), 506-512 (2009). https://doi.org/10.5851/kosfa.2009.29.4.506
  18. Gurtler JB, Beuchat LR.: Performance of Media for Recovering Stressed Cells of Enterobacter sakazakii as Determined Using Spiral Plating and Ecometric Techniques. Appl. Environ. Microbiol., 71(12), 7661-7669 (2005). https://doi.org/10.1128/AEM.71.12.7661-7669.2005
  19. Restino L, Frampton EW, Lionberg WC, Becker RJ.: A Chromogenic Plating Medium for the Isolation and Identification of Enterobacter sakazakii from Foods, Food Ingredients, and Environmental Sources. J. Food Prot., 69(2), 315-322 (2006). https://doi.org/10.4315/0362-028X-69.2.315
  20. Chen YI, Hammack TS, Song KY, Lampel KA.: Evaluation of a revised U.S. Food and Drug Administration method for the detection and isolation of Enterobacter sakazakii in powdered infant formula: precollaborative study. J AOAC Int., 92(3), 862-872 (2009).
  21. Lee JH, Song KY, Hyeon JY, Hwang IG, Kwak HS, 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(3), 410-418 (2010). https://doi.org/10.5851/kosfa.2010.30.3.410
  22. Seo KH, Brackett RE.: Rapid, Specific Detection of Enterobacter sakazakii in Infant Formula Using a Real-Time PCR Assay. J. Food Prot., 68(1), 59-63 (2005). https://doi.org/10.4315/0362-028X-68.1.59
  23. Dominique C, Laurence M, Virginie P, Jose-Carlos F, Chantal B, Sylvain B.: Multilocus sequence analysis of the genus Citrobacter and description of Citrobacter pasteurii sp. nov. Int. J. Syst, Evol. Microbiol., 65(5), 1486-1490 (2015). https://doi.org/10.1099/ijs.0.000122
  24. Hyeon JY, Hwang IG, Kwak HS, Park C, Choi IS, Seo KH.: Evaluation of PCR inhibitory effect of enrichment broths and comparison of DNA extraction methods for detection of Salmonella Enteritidis using real-time PCR assay. J. Vet. Sci., 11(2), 143-149 (2010). https://doi.org/10.4142/jvs.2010.11.2.143
  25. Chon JW, Hyeon JY, Yim JH, Kim JH, Song KY, Seo KH.: Improvement of Modified Charcoal-Cefoperazone-Deoxycholate Agar by Supplementation with a High Concentration of Polymyxin B for Detection of Campylobacter jejuni and C. coli in Chicken Carcass Rinses. Appl. Environ. Microbiol., 78(5), 1624-1626 (2012). https://doi.org/10.1128/AEM.07180-11