Original Articles
Vol. 9 No. 2 (2020)
Detection and quantification of Campylobacter in foods: New analytic approaches to detect and quantify Campylobacter spp. in food samples
Publisher's note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
Received: 30 September 2019
Accepted: 7 January 2020
Accepted: 7 January 2020
1408
Views
748
Downloads
70
HTML
Authors
The aim of the present study was to develop rapid qualitative and quantitative methods based on the use of Real-Time PCR and Droplet Digital PCR (ddPCR), in order to have reliable techniques to detect and quantify Campylobacter spp. in food samples. The gene 16S-rRNA was used as specific target for Campylobacter spp. Real- Time PCR evaluation assay and a not competitive internal control was ushered in it. To investigate the selectivity of the method, 26 Campylobacter strains and 40 non-Campylobacter strains were tested and in order to verify the application of Real-Time PCR method, 5 pork meat samples were experimentally inoculated with a Campylobacter jejuni strain. Subsequently, dilutions with a bacterial load of Campylobacter jejuni within 10-106 CFU/mL were chosen for the optimization of the ddPCR assay. Lastly, a total of 54 naturally contaminated foods samples were analyzed through molecular (Real-Time PCR and ddPCR) and traditional methods. The Real-Time PCR protocol demonstrated to amplify only the Campylobacter spp. strains and when Campylobacter jejuni was experimentally inoculated in meat samples the pathogen was always detected. The ddPCRs assay allowed to quantify a level of contamination of 10 CFU/mL, but it was unable to quantify levels of 105 – 106 CFU/mL. Lastly, Campylobacter spp. was never detected in the 54 samples tested. In conclusion, the novel analytic approach proposed, based on an initial screening of the samples with Real-Time PCR and then on quantification of Campylobacter spp. with a ddPCR on those positive, represents a quick monitoring tool and, if used correctly, it would allow the implementation of food safety.
Downloads
Download data is not yet available.
Citations
Ahmed A, Engelberts MFM, Boer KR, Ahmed N, Hartskeerl RA, 2009. Development and Validation of a Real-Time PCR for Detection of Pathogenic Leptospira Species in Clinical Materials. PLoS One 4:9 pp 4–11. DOI: https://doi.org/10.1371/journal.pone.0007093
EFSA, 2018. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2017. EFSA Journal 2018;16(12):5500.
El-zamkan MA, Hameed KGA, 2016. Prevalence of Campylobacter jejuni and Campylobacter coli in raw milk and some dairy products. Vet. World 9:1147–1151. DOI: https://doi.org/10.14202/vetworld.2016.1147-1151
Erdősi O, Szakmár K, Szili Z, Szita G, Bernáth S, Sövényi J, Laczay P 2018. Rapid in-house detection method of campylobacter spp . from food by redox potential monitoring combined with Real-Time PCR. Acta Vet Hung. 66,1,:1–11. DOI: https://doi.org/10.1556/004.2018.001
European Commission, 2017. Commission Regulation (EU) 2017/1495 of 23 August 2017 amending Regulation (EC) No 2073/2005 as regards Campylobacter in broiler carcases. In: Official Journal of the European Union L 218/1.
Fabiani L, Delibato E, Volpe G, Piermarini S, De Medici D, Palleschi G, 2019. Development of a sandwich ELIME assay exploiting different antibody combinations as sensing strategy for an early detection of Campylobacter. Sensor Actuat B-Chem 290:318-325. DOI: https://doi.org/10.1016/j.snb.2019.03.140
Gobert G, Cotillard A, Fourmestraux C, Pruvost L, Miguet J, Boyer M, 2018. Droplet digital PCR improves absolute quantification of viable lactic acid bacteria in faecal samples. J Microbiol Methods. 148:64–73. DOI: https://doi.org/10.1016/j.mimet.2018.03.004
Hansson I, Sandberg M, Habib I, Lowman R, Engvall EO, 2018. Knowledge gaps in control of Campylobacter for prevention of campylobacteriosis. Transbound Emerg Dis. 65, 1:30–48. DOI: https://doi.org/10.1111/tbed.12870
Hindson BJ, Ness KD, Masquelier DA Belgrader P, Heredia NJ, Makarewicz AJ, Bright IJ, Lucero MY, Hiddessen AL, Legler TC, Kitano TK, Hodel MR, Petersen JF, Wyatt PW, Steenblock ER, Shah PH, Bousse LJ, Troup CB, Mellen JC, Wittmann DK, Erndt NG, Cauley TH, Koehler RT, So AP, Dube S, Rose KA, Montesclaros L, Wang S, Stumbo DP, Hodges SP, Romine S, Milanovich FP, White HE, Regan JF, Karlin-Neumann GA, Hindson CM, Saxonov S, Colston BW, 2011. High-Throughput Droplet Digital PCR System for Absolute Quantitation of DNA Copy Number. Anal Chem. 83,22:8604-10 DOI: https://doi.org/10.1021/ac202028g
ISO,2017.Microbiology of the food chain — Horizontal method for detection and enumeration of Campylobacter spp. — Part 1: Detection method ISO Norm 10272-1:2017. International Standardization Organization ed., Geneva, Switzerland.
ISO, 2017. Microbiology of the food chain — Horizontal method for detection and enumeration of Campylobacter spp. — Part 2: Colony-count technique. ISO Norm 10272-2:2017. International Standardization Organization ed., Geneva, Switzerland.
Karki AB., Marasini D, Oakey CK, Mar K., Fakhr MK, 2018. Campylobacter coli From Retail Liver and Meat Products Is More Aerotolerant Than Campylobacter jejuni. Front Microbiol. 9:2951. DOI: https://doi.org/10.3389/fmicb.2018.02951
Persson S, Eriksson R, Lowther J, Ellström P, Simonsson M, 2018. Comparison between RT droplet digital PCR and RT real-time PCR for quantification of noroviruses in oysters. Int J Food Microbiol. 284:73–83. DOI: https://doi.org/10.1016/j.ijfoodmicro.2018.06.022
Peruzy MF, Murru N, Yu Z, Cnockaert M, Joossens M, Proroga YTR., Houf K 2019. Determination of the microbiological contamination in minced pork by culture dependent and 16S amplicon sequencing analysis. Int J Food Microbiol. 290:27–35. DOI: https://doi.org/10.1016/j.ijfoodmicro.2018.09.025
Peruzy MF, Murru N, Yu Z, Kerkhof P, Neola B, Joossens M, Proroga YTR., Houf K 2019. Assessment of microbial communities on freshly killed wild boar meat by MALDI-TOF MS and 16S rRNA amplicon sequencing. Int J Food Microbiol. 301: 51–60. DOI: https://doi.org/10.1016/j.ijfoodmicro.2019.05.005
Peruzy MF, Murru N, Perugini AG, Capuano F, Delibato E, Mercogliano R, Korkeala H, Proroga YTR 2017. Evaluation of virulence genes in Yersinia enterocolitica strains using SYBR Green real-time PCR. Food Microbiol. 65:231–235. DOI: https://doi.org/10.1016/j.fm.2017.03.004
Porcellato D, Narvhus J, Skeie SB, 2016. Detection and quantification of Bacillus cereus group in milk by droplet digital PCR. J Microbiol Methods. 127: 1–6. DOI: https://doi.org/10.1016/j.mimet.2016.05.012
Ricke SC, Feye KM, Chaney WE., Shi Z, Pavlidis H, Yang Y, 2019. Developments in Rapid Detection Methods for the Detection of Foodborne Campylobacter in the United States. Front Microbiol. 9:3280 DOI: https://doi.org/10.3389/fmicb.2018.03280
Tang H, Cai Q, Li H, Hu P, 2016. Comparison of droplet digital PCR to real-time PCR for quantification of hepatitis B virus DNA. Biosci Biotechnol Biochem. 80,11:2159-2164 DOI: https://doi.org/10.1080/09168451.2016.1196576
Wang M, Yang J, Gai Z, Huo S, Zhu J, Li J, Wang R, Xing S, Shi G, Shi F, Zhang L, 2018. Comparison between digital PCR and real-time PCR in detection of Salmonella typhimurium in milk. International Journal of Food Microbiology. Int J Food Microbiol. 266: 251–256. DOI: https://doi.org/10.1016/j.ijfoodmicro.2017.12.011
Wang Y, Salazar JK, 2016. Culture-Independent Rapid Detection Methods for Bacterial Pathogens and Toxins in Food Matrices. Compr Rev Food Sci F 15 DOI: https://doi.org/10.1111/1541-4337.12175
Yu Z, Peruzy MF, Dumolin C, Joossens M, Houf ,K 2019. Assessment of food microbiological indicators applied on poultry carcasses by culture combined MALDI-TOF MS identification and 16S rRNA amplicon sequencing. Food Microbiol. 82:53-61 DOI: https://doi.org/10.1016/j.fm.2019.01.018
Zhao C, Ge B, Villena JD E, Sudler R, Yeh E, Zhao S, White DG, Wagner D, Meng J., 2001. Prevalence of Campylobacter spp., Escherichia coli, and Salmonella Serovars in Retail Chicken, Turkey, Pork, and Beef from the Greater Washington, D.C., Area. Appl Environ Microbiol. 67(12): 5431–5436. DOI: https://doi.org/10.1128/AEM.67.12.5431-5436.2001
How to Cite
1.
Detection and quantification of Campylobacter in foods: New analytic approaches to detect and quantify Campylobacter spp. in food samples. Ital J Food Safety [Internet]. 2020 Aug. 28 [cited 2025 Nov. 8];9(2). Available from: https://www.pagepressjournals.org/ijfs/article/view/8591
Most read articles by the same author(s)
- Nicoletta Murru, Giuseppe Blaiotta, Maria Francesca Peruzy, Serena Santonicola, Raffaelina Mercogliano, Maria Aponte, Screening of oxalate degrading lactic acid bacteria of food origin , Italian Journal of Food Safety: Vol 6 no. 2 (2017)
- Donatella Nava, Salvatore Capo, Vincenzo Caligiuri, Valerio Giaccone, Loredana Biondi, Gerardo Francesco Vaccaro, Achille Guarino, Federico Capuano, Study of the population dynamics of Listeria monocytogenes and Pseudomonas fluorescens in buffalo mozzarella by means of challenge testing , Italian Journal of Food Safety: Vol. 5 No. 3 (2016)
- Andrea Ariano, Luigi Scarano, Amalia Mormile, Maria Barile, Giuseppe Palma, Nicoletta Murru, Shelf life of anchovy products (Engraulis encrasicolus): evaluation of sensory, microbiological and chemical properties , Italian Journal of Food Safety: Vol. 3 No. 1 (2014)
- Raffaelina Mercogliano, Serena Santonicola, Gennaro Raimo, Maurizio Gasperi, Giampaolo Colavita, Extraction and identification of microplastics from mussels: Method development and preliminary results , Italian Journal of Food Safety: Vol. 10 No. 1 (2021)
- Amalia Mormile, Luigi Scarano, Andrea Ariano, Nicoletta Murru, Lucia Vollano, Aniello Anastasio, Microbial characteristics of Conciato Romano: an artisanal cheese made from raw sheep’s milk , Italian Journal of Food Safety: Vol. 2 No. 3 (2013)
- Nicola Costanzo, Adriano Michele Luigi Santoro, Eleonora Sarno, Antonio Di Loria, Rosa Daniela Grembiale, Domenico Britti, Federico Capuano, Use of sonoelastography to evaluate texture modifications of Mozzarella di Bufala Campana Protected Designation of Origin during storage at different temperatures , Italian Journal of Food Safety: Vol 4, No 4 (2015)
- Serena Santonicola, Maria Francesca Peruzy, Mariagrazia Girasole, Nicoletta Murru, Maria Luisa Cortesi, Raffaelina Mercogliano, Preliminary study on physicochemical and biochemical stress markers at poultry slaughterhouse , Italian Journal of Food Safety: Vol 6 no. 2 (2017)
- Orlandina Di Maro, Yolande T.R. Proroga, Silvia Castellano, Anna Balestrieri, Federico Capuano, Enrico Arletti, Michelangelo Vietina, Melissa Bizzarri, Nicoletta Murru, Maria Francesca Peruzy, Daniela Cristiano, Detection of pathogenic Vibrio spp. in foods: polymerase chain reaction-based screening strategy to rapidly detect pathogenic Vibrio parahaemolyticus, Vibrio cholerae, and Vibrio vulnificus in bivalve mollusks and preliminary results , Italian Journal of Food Safety: Vol. 13 No. 1 (2024)
- Giorgio Smaldone, Raffaele Marrone, Lucia Vollano, Maria Francesca Peruzy, Carmela Maria Assunta Barone, Rosa Luisa Ambrosio, Aniello Anastasio, Microbiological, rheological and physical-chemical characteristics of bovine meat subjected to a prolonged ageing period , Italian Journal of Food Safety: Vol. 8 No. 3 (2019)
- Serena Santonicola, Michela Volgare, Emilia Di Pace, Raffaelina Mercogliano, Mariacristina Cocca, Gennaro Raimo, Giampaolo Colavita, Research and characterization of fibrous microplastics and natural microfibers in pelagic and benthic fish species of commercial interest , Italian Journal of Food Safety: Vol. 12 No. 1 (2023)
