Real-time PCR detection of Campylobacter spp.: A comparison to classic culturing and enrichment

Boer, Paulo de; Rahaoui, H.; Leer, R.J.; Montijn, R.C.; Vossen, J.M.B.M. van der

doi:10.1016/j.fm.2015.05.006

Cited by 59 publications

(43 citation statements)

References 19 publications

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“…The application of qPCR to the food industry supports the detection of pathogens; the approach is recognized and trusted (Boyer and Combrisson ), and it has been used to detect different species of Campylobacter (Postollec and others ; Vencia and others ). For detection of the Campylobacter genus, the primers used are specific for 16 rRNA genes (Garcia and others ; de Boer and others ; Gosselin‐Theberge and others ). However, for detection of C. jejuni , the primers can be specific for the hip O gene (de Boer and others ), the cco N or the hyp O gene (Toplak and others ), the rpo B gene (Rantsiou and others ), or open reading frame (ORF) C (Sails and others ).…”

Section: Molecular Tools Used For Detection Of Campylobacter Spp In mentioning

confidence: 99%

“…For detection of the Campylobacter genus, the primers used are specific for 16 rRNA genes (Garcia and others ; de Boer and others ; Gosselin‐Theberge and others ). However, for detection of C. jejuni , the primers can be specific for the hip O gene (de Boer and others ), the cco N or the hyp O gene (Toplak and others ), the rpo B gene (Rantsiou and others ), or open reading frame (ORF) C (Sails and others ). For detection of C. coli , primers specific for the cad F gene (Toplak and others ) or the gly A gene (de Boer and others ) have been used.…”

Section: Molecular Tools Used For Detection Of Campylobacter Spp In mentioning

confidence: 99%

“…However, for detection of C. jejuni , the primers can be specific for the hip O gene (de Boer and others ), the cco N or the hyp O gene (Toplak and others ), the rpo B gene (Rantsiou and others ), or open reading frame (ORF) C (Sails and others ). For detection of C. coli , primers specific for the cad F gene (Toplak and others ) or the gly A gene (de Boer and others ) have been used. A significant number of studies report using qPCR for detection of Campylobacter spp.…”

Section: Molecular Tools Used For Detection Of Campylobacter Spp In mentioning

confidence: 99%

See 2 more Smart Citations

Molecular Detection, Typing, and Quantification of Campylobacter spp. in Foods of Animal Origin

Frasão

Marin

Conté-Júnior

2017

Comp Rev Food Sci Food Safe

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Abstract:The most frequently reported zoonosis and the main bacterial foodborne disease infection in humans is caused by Campylobacter spp., and C. jejuni and C. coli are the most common types. These bacteria can be found in the intestinal tracts of cattle, dogs, cats, sheep, poultry and pigs. The isolation of this microorganism is laborious because it requires specific media and a low oxygen concentration for growth. Additionally, differentiation between species through conventional bacteriology is difficult, as there are few different biochemical characteristics among the various species. Molecular microbiological techniques have become more important and are now broadly applied to help overcome difficulties in the identification, differentiation, and quantification of this pathogen. To date, there have been advances in the development and use of molecular techniques for the identification of microorganisms in foodstuffs. Tools such as pulsed-field gel electrophoresis and multilocus sequence typing are the most commonly used for typing. For the identification and confirmation of species, polymerase chain reaction (PCR) is crucial. Quantification by real-time PCR has wide applicability. To identify strains and antimicrobial resistance genes, sequencing technologies have been applied. This review builds on the discussion about the main and most widely used molecular methods for Campylobacter, as well as methods showing better potential for the classification, identification, and quantification of this important pathogen.

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Section: Molecular Tools Used For Detection Of Campylobacter Spp In mentioning

confidence: 99%

Section: Molecular Tools Used For Detection Of Campylobacter Spp In mentioning

confidence: 99%

Section: Molecular Tools Used For Detection Of Campylobacter Spp In mentioning

confidence: 99%

See 1 more Smart Citation

Molecular Detection, Typing, and Quantification of Campylobacter spp. in Foods of Animal Origin

Frasão

Marin

Conté-Júnior

2017

Comp Rev Food Sci Food Safe

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“…Detection of Campylobacter spp. in feces by direct PCR (direct detection of target DNA from samples) [75] or by PCR after a selective enrichment step (detection of target DNA from enrichment broth culture incubated at 37 °C for 4 h followed by 48 h at 42 °C ), has been applied successfully to human, avian and animal feces [76][77][78]. Determination of campylobacters in feces by PCR provides more accurate account of the incidence of the organism, furthermore, PCR potentially reduces the time for detection and eliminates the need for conventional confirmatory methods [5,73].…”

Section: Detection In Fecesmentioning

confidence: 99%

Campylobacter in the environment: A major threat to public health

Abulreesh¹,

Organji²,

Elbanna³

et al. 2017

APJTD

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“…A survey of Campylobacter in broilers at slaughter across the EU reported that 60.4% of colonised flocks carried C. jejuni and the remainder carried C. coli (Anonymous, ). This survey only identified a single Campylobacter colony per flock and could not account for the scenario where flocks are co‐colonised with both species (Arnold et al., ; de Boer et al., ).…”

Section: Introductionmentioning

confidence: 99%

Sensitivity of Direct Culture, Enrichment and PCR for Detection of Campylobacter jejuni and C. coli in Broiler Flocks at Slaughter

Rodgers

Simpkin

Lee

et al. 2016

Zoonoses and Public Health

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Broiler chicken flocks are a significant source of Campylobacter jejuni and Campylobacter coli that result in the major public health problem of campylobacteriosis. Accurate estimates of the prevalence of both C. coli and C. jejuni in flocks would enhance epidemiological understanding, risk assessment and control options. This study combined results from a panel of 10 detection tests (direct culture, enrichment and PCR) on caecal samples from flocks at slaughter. A parallel interpretation approach was used to determine the presence of Campylobacter spp. and for C. jejuni and C. coli individually. The sample was considered positive if at least one method detected the target and this interpretation was taken to represent a 'proxy gold standard' for detection in the absence of a gold standard reference test. The sensitivity of each individual method to detect Campylobacter spp., C. jejuni and C. coli was then estimated relative to the proxy gold standard. Enrichment in adapted Exeter broth (deficient in polymyxin B) with a resuscitation step was 100% sensitive, whilst direct culture on modified charcoal cefoperazone deoxycholate agar (mCCDA) was highly sensitive (97.9%). Enrichment methods using Preston broth and Bolton broth were significantly less sensitive. Enrichment in Exeter broth promoted the recovery of C. jejuni, whilst enrichment in Bolton broth favoured C. coli. A RT-PCR detection test could identify 80% of flocks that were co-colonised with both species. This study found that 76.3% (n = 127) of flocks were colonised with Campylobacter spp. The majority (95.9%) of Campylobacter-positive flocks were colonised with C. jejuni; however, approximately one-third of positive flocks were simultaneously colonised with both C. jejuni and C. coli. The findings highlight the impact of different detection methodologies on the accuracy of the estimated incidence of both C. jejuni and C. coli entering the abattoir within broiler flocks and the associated public health risks.

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Real-time PCR detection of Campylobacter spp.: A comparison to classic culturing and enrichment

Cited by 59 publications

References 19 publications

Molecular Detection, Typing, and Quantification of Campylobacter spp. in Foods of Animal Origin

Molecular Detection, Typing, and Quantification of Campylobacter spp. in Foods of Animal Origin

Campylobacter in the environment: A major threat to public health

Sensitivity of Direct Culture, Enrichment and PCR for Detection of Campylobacter jejuni and C. coli in Broiler Flocks at Slaughter

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