Real-time PCR assay for rapid detection and quantification of Campylobacter jejuni on chicken rinses from poultry processing plant

Debretsion, Aradom; Habtemariam, Tsegaye; Wilson, Saul; Nicholas, David; Yehualaeshet, Teshome

doi:10.1016/j.mcp.2006.10.006

Cited by 40 publications

(23 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Such an approach has already been employed for other pathogens like Listeria monocytogenes (Rantsiou et al, 2008). Methods to detect and quantify C. jejuni in foods until recently have been applied after an enrichment step (Josefsen et al, 2004;Nogva et al, 2000;Oliveira et al, 2005;Sails et al, 2003;Yang et al, 2003), while direct quantification of C. jejuni in chicken rinses has been performed (Debretsion et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Survey of Campylobacter jejuni in retail chicken meat products by application of a quantitative PCR protocol

Rantsiou

Lamberti

Cocolin

2010

International Journal of Food Microbiology

View full text Add to dashboard Cite

AbtractCampylobacter-contaminated food products are currently the cause of the highest number of gastroenteritis cases in developed countries. Apart for biosafety measures at the primary production level, no other official control measures are currently in place for its control. This is partly due to the lack of quantitative data regarding the prevelance and contamination level of different food products by Campylobacter spp.that does not allow for quantitative risk assessment. PCR-based methods, applied without prior enrichment, in food samples circumvent limitations associated with the quantification of foodborne pathogens by traditional, culture-dependent methods. In this study, we report the development of a protocol, based on the amplification of the rpoB gene of Campylobacter jejuni, by quantitative PCR (qPCR), directly in food samples. The quantification limit of the protocol was determined to be in the order of 10 colony forming units (cfu)/g or ml of food sample. The optimized protocol was applied for the survey of C. jejuni in naturally contaminated poultry samples. In parallel, traditional sampling was also performed. A high percentage of samples (87%) resulted to be positive by qPCR, while no C. jejuni was detected by traditional analysis. Furthermore, important differences were observed in the detection by qPCR between samples before and after enrichment.

show abstract

Section: Introductionmentioning

confidence: 99%

Survey of Campylobacter jejuni in retail chicken meat products by application of a quantitative PCR protocol

Rantsiou

Lamberti

Cocolin

2010

International Journal of Food Microbiology

View full text Add to dashboard Cite

show abstract

“…The introduction of real-time quantitative PCR (Q-PCR) has enabled faster, more sensitive, and less labor-intensive quantitative detection. Q-PCR methods for food-borne Campylobacter jejuni and C. coli in poultry, which is recognized as an important source of human Campylobacter infections, have been published (11,12,15,38,46). However, since control strategies mostly focus on reduction of the number of bacterial cells on the chicken carcass, the usefulness of these Q-PCR methods for risk assessment could be limited, since they detect all of the Campylobacter bacteria present in a sample, including the dead cells.…”

mentioning

confidence: 99%

Rapid Quantification of Viable Campylobacter Bacteria on Chicken Carcasses, Using Real-Time PCR and Propidium Monoazide Treatment, as a Tool for Quantitative Risk Assessment

Josefsen

Löfström

Hansen

et al. 2010

Appl Environ Microbiol

159

134

View full text Add to dashboard Cite

“…using phenotypic methods is difficult because of their low biochemical activity, fastidious nature, and frequent variability in the results obtained (On, 1996). Molecular methods and PCR-based techniques, in particular, are being increasingly applied to detect and quantify food-borne pathogens (Debretsion et al, 2007;Park et al, 2011;Alves et al, 2012;Ryu et al, 2013). As conventional PCR methods have several disadvantages, including cost, time-consuming separation of the PCR products by gel electrophoresis, and inability to analyze large number of samples, the real-time PCR technique provides rapid, highly sensitive, and specific identification.…”

Section: Pcr Optimizationmentioning

confidence: 99%

“…The newly developed method is capable of detecting 1 genomic copy per reaction, which is comparable with real-time PCR assays described by other authors. Debretsion et al (2007) reported a detection limit of 1 genome copy for TaqMan Campylobacter PCR assay, whereas Park et al (2011) showed a sensitivity of 7.33 × 10 1 copies for pure culture C. jejuni in a uniplex SYBR Green real-time PCR. We optimized various conditions of the amplification process, focusing on the annealing temperature and the concentration of primers to obtain the lowest C T value during amplification of C. jejuni DNA.…”

Section: Pcr Optimizationmentioning

confidence: 99%

“…Compared with the conventional phenotyping methods for pathogen identification, real-time PCR is a sensitive, accurate, and rapid method, and does not require postamplification steps that eliminate the risk of cross-contamination (Barletta et al, 2013). Thus far, numerous studies have been published to estimate the prevalence and concentration of Campylobacter in poultry samples during processing using different approaches (Rosenquist et al, 2006;Debretsion et al, 2007;Maćkiw et al, 2008;Hue et al, 2010;Habib et al, 2012;Trimble et al, 2013). As far as we are aware, no previous study has reported on the application of a simple and rapid real-time PCR method to allow sensitive and highly specific identification of C. jejuni at different stages of poultry processing.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Rapid identification of Campylobacter jejuni from poultry carcasses and slaughtering environment samples by real-time PCR

et al. 2014

View full text Add to dashboard Cite

The objective of this study was to develop a real-time PCR assay for rapid identification of Campylobacter jejuni and to apply the method in analyzing samples from poultry processing. A C. jejuni-specific primer set targeting a portion of the C. jejuni hippuricase gene was developed. The specificity of the newly designed primer pair was verified using 5 C. jejuni strains and 20 other bacterial strains. Sensitivity was determined to be as low as 1 genome copy per reaction. A total of 73 samples were collected at different sites along the processing line during 2 visits to a poultry slaughterhouse and were examined by direct plating onto modified charcoal cefoperazone deoxycholate agar or after enrichment in Bolton broth followed by plating on modified charcoal cefoperazone deoxycholate agar. The newly developed real-time PCR assay was used to identify the presumptive colonies as belonging to C. jejuni. A real-time PCR assay targeting 16S ribosomal RNA was also applied to determine Campylobacter spp. prevalence. Results from the real-time PCR analysis indicated considerable variability in Campylobacter contamination, with incidence rates of 72.7 and 27.6% for sampling days A and B, respectively. Campylobacter was isolated from 100% of prescalded and preeviscerated carcasses on sampling day A. In contrast, on sampling day B, the highest number of Campylobacter-positive carcasses was recovered after evisceration (60%). The chilling process significantly reduced (P < 0.05) Campylobacter population, but the percentage of positive samples on sampling day A increased to 80%. All samples collected from the processing environment, except scalding tank 3 and the prechiller and chiller tanks, were 100% positive on day A, whereas no campylobacters were isolated from machinery on sampling day B. Our results revealed the widespread of C. jejuni in poultry processing and proved that the newly developed real-time PCR assay is a simple, specific, and inexpensive method for rapid C. jejuni identification. The newly developed PCR method can be easily used in laboratories for reliable and unambiguous identification of C. jejuni in poultry samples.

show abstract

Real-time PCR assay for rapid detection and quantification of Campylobacter jejuni on chicken rinses from poultry processing plant

Cited by 40 publications

References 21 publications

Survey of Campylobacter jejuni in retail chicken meat products by application of a quantitative PCR protocol

Survey of Campylobacter jejuni in retail chicken meat products by application of a quantitative PCR protocol

Rapid Quantification of Viable Campylobacter Bacteria on Chicken Carcasses, Using Real-Time PCR and Propidium Monoazide Treatment, as a Tool for Quantitative Risk Assessment

Rapid identification of Campylobacter jejuni from poultry carcasses and slaughtering environment samples by real-time PCR

Contact Info

Product

Resources

About