0.917
IF5
1.024
IF
Q2
JCR
0.90
CiteScore
0.385
SJR
Q2
SJR
20
MNiSW
142.18
ICV
ORIGINAL PAPER
 
CC-BY 4.0
 
 

Quantitative assessment of Yersinia enterocolitica in raw pork meat using real time PCR (qPCR) technique

 
1
Lomza State University of Applied Sciences, Akademicka 14, 18-400 Łomża
J. Anim. Feed Sci. 2017;26(2):141–147
Publish date: 2017-06-14
KEYWORDS:
TOPICS:
ABSTRACT:
The aim of the present study was to develop a highly sensitive and specific TaqMan probe with a set of primers for detection of pathogenic Yersinia enterocolitica strains and to eliminate the pre-PCR enrichment step in the realtime PCR (qPCR). A newly developed qPCR assay which is sensitive and specific for quick and reliable quantitative assessment of Y. enterocolitica present in artificially contaminated raw pork meat samples is described. This protocol involves the qPCR method with a TaqMan probe. The primers and probe were designed on the base of locus_tag CH49_3099 gene. This protocol appeared to be reliable for both intended applications: 1. identification and quantification of Y. enterocolitica in artificially and naturally contaminated raw pork meat and 2. establishment of growth potentials of different serotypes of Y. enterocolitica in raw meat at the usually used storage temperatures. This developed method makes it possible to eliminate the pre-PCR enrichment step enabling for the rapid assessment of meat-related consumer exposure to this pathogen.
CORRESPONDING AUTHOR:
M. A. Stachelska   
Lomza State University of Applied Sciences, Akademicka 14, 18-400 Łomża
 
REFERENCES (21):
1. Bolton D.J., Ivory C., McDowell D., 2013. A small study of Yersinia enterocolitica in pigs from birth to carcass and characterization of porcine and human strains. Food Control 33, 521–524, https://doi.org/10.1016/j.foodcont.2013.03.039.
2. Bonardi S., Alpigiani I., Pongolini S., Morganti M., Tagliabue S., Bacci C., Brindani F., 2014. Detection, enumeration and characterization of Yersinia enterocolitica 4/O:3 in pig tonsils at slaughter in Northern Italy. Int. J. Food Microbiol. 177, 9–15, https://doi.org/10.1016/j.ijfoodmicro.2014.02.005.
3. Boyapalle S., Wesley I.V., Hurd H.S., Reddy P.G., 2001. Comparison of culture, multiplex, and 5’ nuclease polymerase chain reaction assay for the rapid detection of Yersinia enterocolitica in swine and pork products. J. Food Protect. 64, 1352–1361, https://doi.org/10.4315/0362-028X-64.9.1352.
4. EC, 2006. Guidance Document of the Commission of the European Communities on official controls, under Regulation (EC) No. 882/2004, concerning microbiological sampling and testing of foodstuffs.
5. Fosse J., Seegers H., Magras C., 2008. Foodborne zoonoses due to meat: a quantitative approach for a comparative risk assessment applied to pig slaughtering in Europe. Vet. Res. 39, 01, https://doi.org/10.1051/vetres:2007039.
6. Fredriksson-Ahomaa M., Korkeala H., 2003. Low occurrence of pathogenic Yersinia enterocolitica in clinical, food, and environmental samples: a methodological problem. Clin. Microbiol. Rev. 16, 220–229, https://doi.org/10.1128/CMR.16.2.220-2003.
7. Fredriksson-Ahomaa M., Stolle A., Stephan R., 2007. Prevalence of pathogenic Yersinia enterocolitica in pigs slaughtered at a Swiss abattoir. Int. J. Food Microbiol. 119, 207–212, https://doi.org/10.1016/j.ijfoodmicro.2007.07.050.
8. Galindo C.L., Rosenzweig J.A., Kirtley M.L., Chopra A.K., 2011. Pathogenesis of Y. enterocolitica and Y. pseudotuberculosis in human yersiniosis. J. Pathog. 2011, 182051, https://doi.org/10.4061/2011/182051.
9. Gürtler M., Alter T., Kasimir S., Linnebur M., Fehlhaber K., 2005. Prevalence of Yersinia enterocolitica in fattening pigs. J. Food Protect. 68, 850-854, https://doi.org/10.4315/0362-028X-68.4.850.
10. Martinez P.O., 2010. Prevalence of enteropathogenic Yersinia in pigs from different European countries and contamination in pork production chain. MA Dissertation. Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of Helsinki (Finland), http://urn.fi/URN:ISBN:978-952-10-6393-0.
11. Najdenski H., Heyndrickx M., Herman L., Werbrouck H., Van Coillie E., 2012. Quantification of Yersinia enterocolitica in raw milk using qPCR. Vet. Microbiol. 160, 428–434, https://doi.org/10.1016/j.vetmic.2012.06.013.
12. Stenkova A.M., Isaeva M.P., Rasskazov V.A., 2008. Development of a multiplex PCR procedure for detection of Yersinia genus with identification of pathogenic species (Y. pestis, Y. pseudotuberculosis, and Y. enterocolitica). Mol. Genet. Microbiol. Virol. 23, 119–125, https://doi.org/10.3103/S0891416808030038.
13. Tauxe R.V., Wauters G., Goossens V., Van Noyen R., Vandepitte J., Martin S.M., De Mol P., Thiers G., 1987. Yersinia enterocolitica infections and pork. Lancet 329(8542), 1129–1132, https://doi.org/10.1016/S0140-6736(87)91683-7.
14. Thisted Lambertz S., Granath K., Fredriksson-Ahomaa M., Johansson K.-E., Danielsson-Tham M.-L., 2007. Evaluation of a combined culture and PCR method (NMKL-163A) for detection of presumptive pathogenic Yersinia enterocolitica in pork products. Journal of Food Protection 70 (2), 335–340, https://doi.org/10.4315/0362-028X-70.2.335.
15. Thisted Lambertz S., Nilsson C., Hallanvuo S., Lindblad M., 2008. Real-time PCR method for detection of pathogenic Yersinia enterocolitica in food. Appl. Environ. Microbiol. 74, 6060–6067, https://doi.org/10.1128/AEM.00405-08.
16. Van Damme I., Berkvens D., Baré J., De Zutter L., 2013a. Influence of isolation methods on the occurrence of plasmid-carrying Yersinia enterocolitica serotype O:3 in slaughter pig tonsils, faeces and carcass surface swabs. Int. J. Food Microbiol. 164, 32–35, https://doi.org/10.1016/j.ijfoodmicro.2013.03.019.
17. Van Damme I., Berkvens D., Botteldoorn N., Dierick K., Wits J., Pochet B., De Zutter L., 2013b. Evaluation of the ISO 10273:2003 method for the isolation of human pathogenic Yersinia enterocolitica from pig carcasses and minced meat. Food Microbiol. 36, 170-175, https://doi.org/10.1016/j.fm.2013.05.007.
18. Van Damme I., Habib I., De Zutter L., 2010. Yersinia enterocolitica in slaughter pig tonsils: Enumeration and detection by enrichment versus direct plating culture. Food Microbiol. 27, 158-161, https://doi.org/10.1016/j.fm.2009.09.011.
19. Vázlerová M., Steinhauserová I., 2006. The comparison of the methods for the identification of pathogenic serotypes and biotypes of Yersinia enterocolitica: Microbiological methods and PCR. Czech J. Food Sci. 24, 217–222.
20. Wauters G., Kandolo K., Janssens M., 1987. Revised biogrouping scheme of Yersinia enterocolitica. Contrib. Microbiol. Immunol. 9, 14–21.
21. Weynants V., Jadot V., Denoel P.A., Tibor A., Letesson J.J., 1996. Detection of Yersinia enterocolitica serogroup O:3 by a PCR method. J. Clin. Microbiol. 34, 1224–1227.
ISSN:1230-1388