Evaluation of a PCR assay for the detection and identification of Campylobacter jejuni and Campylobacter coli in retail poultry products

2005 ◽  
Vol 156 (4) ◽  
pp. 568-574 ◽  
Author(s):  
Estibaliz Mateo ◽  
Jose Cárcamo ◽  
Maria Urquijo ◽  
Ildefonso Perales ◽  
Aurora Fernández-Astorga
Keyword(s):  
Campylobacter Jejuni ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
Poultry Products ◽  
Detection And Identification

Incidence of Salmonella, Campylobacter jejuni, Campylobacter coli, and Listeria monocytogenes in Poultry Carcasses and Different Types of Poultry Products for Sale on the Belgian Retail Market

1999 ◽  
Vol 62 (7) ◽  
pp. 735-740 ◽  
Author(s):  
M. UYTTENDAELE ◽  
P. DE TROY ◽  
J. DEBEVERE
Keyword(s):  
Listeria Monocytogenes ◽  
Campylobacter Jejuni ◽  
Broiler Chickens ◽  
Salmonella Enteritidis ◽  
Contamination Level ◽  
Contamination Rate ◽  
Campylobacter Coli ◽  
Retail Market ◽  
Salmonella Spp ◽  
Poultry Products

From January 1997 to May 1998, 772 samples of poultry carcasses and poultry products for sale on the retail market in Belgium were analyzed for the presence of Salmonella spp., Salmonella Enteritidis, Campylobacter jejuni, C. coli, and Listeria monocytogenes per 100 cm2 or 25 g. Poultry samples were contaminated with Salmonella (36.5%), C. jejuni and C. coli (28.5%), and L. monocytogenes (38.2%). In about 12.3% of the poultry samples, the L. monocytogenes contamination level exceeded 1 CFU per g or cm2. Significant differences in pathogen contamination rates of poultry products were noticed between the poultry products originating from Belgian, French, and U.K. abattoirs. Poultry products derived from broiler chickens running free in pine woods until slaughtering age (12 to 13 weeks) had a significantly (P < 0.05) lower contamination rate of Salmonella than poultry products from enclosed broilers slaughtered at the age of 6 to 8 weeks. A significantly (P < 0.05) lower pathogen contamination rate was noted for Salmonella, C. jejuni, and C. coli for poultry cuts without skin compared to poultry cuts with skin on. An increase in pathogen contamination rate was noticed during cutting and further processing. To diminish C. jejuni, C. coli, Salmonella, and L. monocytogenes contamination rates, hygienic rules of slaughter and meat processing must be rigorously observed. At the moment, zero tolerance for these pathogens is not feasible, and there is a need to establish criteria allowing these pathogens to be present at reasonable levels in the examined poultry samples.

scholarly journals Detection of Small Numbers of Campylobacter jejuni and Campylobacter coli Cells in Environmental Water, Sewage, and Food Samples by a Seminested PCR Assay

1999 ◽  
Vol 65 (4) ◽  
pp. 1636-1643 ◽  
Author(s):  
Astrid S. Waage ◽  
Traute Vardund ◽  
Vidar Lund ◽  
Georg Kapperud
Keyword(s):  
Campylobacter Jejuni ◽  
Simple Procedure ◽  
Environmental Water ◽  
Food Samples ◽  
Proteinase K ◽  
Coliform Bacteria ◽  
Intergenic Sequence ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
Seminested Pcr

ABSTRACT A rapid and sensitive assay was developed for detection of small numbers of Campylobacter jejuni and Campylobacter coli cells in environmental water, sewage, and food samples. Water and sewage samples were filtered, and the filters were enriched overnight in a nonselective medium. The enrichment cultures were prepared for PCR by a rapid and simple procedure consisting of centrifugation, proteinase K treatment, and boiling. A seminested PCR based on specific amplification of the intergenic sequence between the two Campylobacter flagellin genes, flaA andflaB, was performed, and the PCR products were visualized by agarose gel electrophoresis. The assay allowed us to detect 3 to 15 CFU of C. jejuni per 100 ml in water samples containing a background flora consisting of up to 8,700 heterotrophic organisms per ml and 10,000 CFU of coliform bacteria per 100 ml. Dilution of the enriched cultures 1:10 with sterile broth prior to the PCR was sometimes necessary to obtain positive results. The assay was also conducted with food samples analyzed with or without overnight enrichment. As few as ≤3 CFU per g of food could be detected with samples subjected to overnight enrichment, while variable results were obtained for samples analyzed without prior enrichment. This rapid and sensitive nested PCR assay provides a useful tool for specific detection of C. jejuni or C. coli in drinking water, as well as environmental water, sewage, and food samples containing high levels of background organisms.

scholarly journals Detection of Campylobacter jejuni, Campylobacter coli, and virulence genes in poultry products marketed in Northeastern Brazil

2021 ◽  
Vol 10 (10) ◽  
pp. e542101019224
Author(s):  
Felipe Pereira de Melo ◽  
Priscila Oliveira da Silva ◽  
Saruanna Millena dos Santos Clemente ◽  
Renata Pimentel Bandeira de Melo ◽  
José Givanildo da Silva ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Virulence Genes ◽  
Northeastern Brazil ◽  
Campylobacter Coli ◽  
Public Markets ◽  
Poultry Products ◽  
Public Market

In this study, we evaluated the prevalence of Campylobacter jejuni, Campylobcater coli, and virulence genes in fresh, chilled, and frozen chicken carcasses with livers and gizzards sold in public markets and supermarkets. Of the 90 samples analyzed, C. jejuni was the most prevalent, with 28.8% of positive samples, whereas C. coli was positive in 15.6% of samples. In public market samples, C. coli had a higher prevalence than C. jejuni, with 16.7% positive samples detected, whereas in supermarket samples, C. jejuni was more prevalent (36.7% positivity). C. jejuni was detected in all forms of commercialized carcasses; however, there was a higher prevalence (43.3%) in chilled samples than C. coli, which was not detected in frozen samples but showed a higher prevalence (16.7%) in fresh samples. Both species were detected in different poultry products, with C. jejuni being more prevalent (53.3%) in liver samples. C. coli showed a higher prevalence in samples of meat pieces (10%). The presence of five virulence genes related to adherence (Peb1, JlpA, CadF, and CapA) and invasion (CiaB) was also observed in both species.

scholarly journals Evaluation of a Multiplex PCR Assay for the Identification of Campylobacter jejuni and Campylobacter coli

2017 ◽  
Vol 3 (1) ◽  
pp. 6-8
Author(s):  
Saeed Shams ◽  
Mehdi Ghorbanalizadgan ◽  
Somayeh Haj Mahmmodi ◽  
Alessandra Piccirillo ◽  
◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Campylobacter Jejuni ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

scholarly journals Multiplex PCR Assay for Identifi cation and Differentiation of Campylobacter jejuni and Campylobacter coli Isolates

Folia Medica ◽  
2016 ◽  
Vol 58 (2) ◽  
pp. 95-100 ◽  
Author(s):  
Maria R. Pavlova ◽  
Elina G. Dobreva ◽  
Katucha I. Ivanova ◽  
Galina D. Asseva ◽  
Ivan N. Ivanov ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Campylobacter Jejuni ◽  
Accurate Method ◽  
Clinical Isolates ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
Biochemical Tests ◽  
Multiplex Pcr Assay ◽  
Hydrolysis Of ◽  
Campylobacter Spp

AbstractIntroduction: Campylobacter spp. are important causative agents of gastrointestinal infections in humans. The most frequently isolated strains of this bacterial genus are Campylobacter jejuni and Campylobacter coli. To date, genetic methods for bacterial identification have not been used in Bulgaria. We optimized the multiplex PSR assay to identify Campylobacter spp. and differentiate C. jejuni from C. coli in clinical isolates. We also compared this method with the routinely used biochemical methods.Aim: To identify Campylobacter spp. and discriminate C. coli from C. jejuni in clinical isolates using multiplex PCR assay.Materials and methods: Between February 2014 and January 2015 we studied 93 stool samples taken from patients with diarrheal syndrome and identified 40 species of Campylobacter spp. in them. The clinical material was cultured in microaerophilic atmosphere, the isolated strains being biochemically diff erentiated (hydrolysis of sodium hippurate for C. jejuni, and hydrolysis of indoxyl acetate for C. coli). DNA was isolated from the strains using QiaAmp MiniKit (QIAGEN, Germany). Twenty strains were tested with multiplex PCR for the presence of these genes: cadF, characteristic for Campylobacter spp., hipO for C. jejuni and asp for C. coli.Results and discussion: The biochemical tests identified 16 strains of C. jejuni, 3 strains of C. coli, and 1 strain of C. upsaliensis. After the multiplex PCR assay the capillary gel electrophoresis confirmed 16 strains of C. jejuni, 2 strains of C. coli and 2 strains of Campylobacter spp. - because of the presence of the gene cadF. C. jejuni has the gene hipO, and it is possible that this gene may not be expressed in the biochemical differentiation yielding a negative reaction as a result. In comparison, we can conclude that the genetic differentiation is a more accurate method than the biochemical tests.Conclusion: The multiplex PCR assay is a fast, accurate method for identifi cation of Campylobacter spp. which makes it quite necessary in the clinical diagnostic practice.

scholarly journals Evaluation of a Multiplex PCR Assay for the Identification of Campylobacter jejuni and Campylobacter coli

2017 ◽  
Vol 3 (1) ◽  
pp. 6-8 ◽  
Author(s):  
Saeed Shams ◽  
◽  
Mehdi Ghorbanalizadgan ◽  
Somayeh Haj Mahmmodi ◽  
Alessandra Piccirillo ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Campylobacter Jejuni ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

scholarly journals Evaluation of Diagnostic Performance of BD Max EBP Assay in Patients with Diarrheal Illness.

2021 ◽  
Author(s):  
Ozlem KOYUNCU OZYURT ◽  
Imran SAGLIK ◽  
Betil OZHAK ◽  
Derya MUTLU ◽  
Belkıs LEVENT ◽  
...  
Keyword(s):  
Acute Diarrhea ◽  
Diagnostic Performance ◽  
Detection Methods ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
Chi Square ◽  
Diarrheal Illness ◽  
Percent Agreement ◽  
Detection And Identification ◽  
Chi Square Test

Abstract Background: Detection of the etiological agents in patients with acute diarrhea is challenging due to a wide variety of pathogens. Detection and identification of the pathogens of acute diarrhea are important for both individual patient care and public health. The aim of this study is to evaluate the diagnostic performance of BD Max Enteric Bacterial Pathogens (EBP) PCR assay in patients with diarrheal illness. Methods: Between 1 January 2014 and 31 May 2015, one thousand two hundred twenty four stool samples from pediatric or adult patients with diarrhea submitted for routine analysis of bacterial stool pathogens were included in the study. We compared the BD Max EBP PCR assay to culture for the detection of Salmonella spp., Shigella spp., Campylobacter jejuni, and Campylobacter coli and an EIA (Enzym Immun Assay) for Shiga toxins 1 and 2. Discordant results were adjudicated by either antigen detection methods or Film array GI (Gastro Intestinal) Panel. Coinfections were excluded. The chi-square test was used for comparisons of PPA and NPA.Results: The PPA (positive percent agreement) values for the BD Max EBP assay was 100% and NPA (negative percent agreement) was between 98.0%-99.7%, when compared with culture and EIA. After discrepant analysis, the PPA values for the BD Max EBP assay was 100% and NPA was between 99.5%-100%. Conclusion: The BD Max EBP assay showed a high correlation rate with conventional and molecular methods for the detection of stool pathogens.

scholarly journals Development of a multiplex PCR assay for identification of Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalis subsp. hyointestinalis, Campylobacter jejuni, Campylobacter lari and Campylobacter upsaliensis

2007 ◽  
Vol 56 (11) ◽  
pp. 1467-1473 ◽  
Author(s):  
Wataru Yamazaki-Matsune ◽  
Masumi Taguchi ◽  
Kazuko Seto ◽  
Ryuji Kawahara ◽  
Kentaro Kawatsu ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Campylobacter Jejuni ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
Specific Pcr ◽  
Campylobacter Fetus ◽  
Multiplex Pcr Assay ◽  
Pcr Products ◽  
Campylobacter Lari ◽  
Campylobacter Upsaliensis

A multiplex PCR assay has been developed for the identification of the six common Campylobacter taxa associated with human gastroenteritis and/or septicaemia, namely Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalis subsp. hyointestinalis, Campylobacter jejuni, Campylobacter lari and Campylobacter upsaliensis. The assay was developed using a combination of newly designed and published primers. It provided a specific PCR product for each of the five Campylobacter species and the one subspecies, and each of the PCR products was sufficiently distinguished by a difference in size by agarose gel electrophoresis. On evaluation of efficacy with 142 Campylobacter strains, the assay correctly identified all strains as 1 of the 6 Campylobacter taxa. This multiplex PCR assay is a rapid, simple and practical tool for identification of the six Campylobacter taxa commonly associated with gastroenteritis and/or septicaemia in humans, and offers an effective alternative to conventional biochemical-based assays.

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