Development of a m-PCR assay for simultaneous identification of Campylobacter jejuni and C. coli

Denis, M.; Soumet, Christophe; Rivoal, Katell; Ermel, Gwennola; Blivet, D.; Salvat, Gilles; Colin, P.

doi:10.1046/j.1472-765x.1999.00658.x

Cited by 313 publications

(222 citation statements)

References 16 publications

Supporting

Mentioning

212

Contrasting

Unclassified

Order By: Relevance

“…For Salmonella spp., the isolated strain was analyzed by the riboprinting method described by De Cesare et al (2001); for C. jejuni and C. coli, the bacterial DNA was extracted and purified using the NucleoSpin Tissue kit (Mecherey-Nagel, Dü ren, Germnay) as described by the manufacturer, and a multiplex PCR protocol described by Denis et al (1999) was used. For VTEC O157:H7, the serotype and the presence of stx1, stx2, and eae genes in E. coli O157: H7 isolate were confirmed by PCR as described above.…”

Section: Laboratory Investigationsmentioning

confidence: 99%

Sale of Raw Milk in Northern Italy: Food Safety Implications and Comparison of Different Analytical Methodologies for Detection of Foodborne Pathogens

Giacometti

Serraino

Finazzi

et al. 2012

Foodborne Pathogens and Disease

View full text Add to dashboard Cite

The safety of raw milk sold in Northern Italy was investigated in relation to hygiene quality parameters and presence of Salmonella spp., Listeria monocytogenes, thermotolerant Campylobacter, and Verocytotoxin producing Escherichia coli O157:H7. The performance of different analytical methods used-official culture method (ISO), modified Bacteriological Analytical Manual cultural method (mBAM), and polymerase chain reaction (PCR)-was evaluated. The presence of Mycobacterium avium subsp. paratuberculosis (Map) was investigated only by PCR. All samples met regulations for alkaline phosphatase and inhibitory substance, while 18% and 44.8% of samples collected from vending machines had, respectively, somatic cell count (SCC) > 300,000/mL and total bacterial count (TBC) > 50,000 CFU/mL. The correlation between hygienic quality parameters in samples collected from bulk tank and vending machines showed a significant increase of TBC in vending machines meaning that raw milk was mishandled during distribution and sale. All pathogens investigated were detected in raw milk sold at vending machines; a total of five samples (5%) had at least one pathogen, of which two were detected by PCR and three by mBAM. None of the samples was positive by cultural ISO methods. Even if the comparison of analytical methods showed that none performs significantly better than the others, testing a higher volume of milk (25 versus 210 mL) affects significantly the detection rate of pathogens. Three samples (3%) were positive for Map, suggesting that raw milk is a significant source of Map exposure for consumers. The observed TBC increase and the detection of several pathogenic bacteria pose questions on the safety of raw milk; the use of ISO seems inefficient in detecting a low contamination level of pathogens in milk and consequently not appropriate as official method for testing. In order to ensure consumer's safety, a new approach for the raw milk chain is required.

show abstract

Section: Laboratory Investigationsmentioning

confidence: 99%

Sale of Raw Milk in Northern Italy: Food Safety Implications and Comparison of Different Analytical Methodologies for Detection of Foodborne Pathogens

Giacometti

Serraino

Finazzi

et al. 2012

Foodborne Pathogens and Disease

View full text Add to dashboard Cite

show abstract

“…In particular, it is often difficult to differentiate between C. jejuni and C. coli by biochemical methods (24). Hippurate hydrolysis activity is a sole biochemical marker for discrimination between C. jejuni and C. coli (5,22). Owing to very weak hippuricase activity of some C. jejuni strains, hippurate-negative strains of C. jejuni are also well recognized (32).…”

mentioning

confidence: 99%

“…Owing to very weak hippuricase activity of some C. jejuni strains, hippurate-negative strains of C. jejuni are also well recognized (32). Several PCR-based assays have been developed to facilitate the differentiation of C. jejuni from C. coli (4,5,9,15,18,24,33), however the ability of these assays to accurately identify Campylobacter species has not yet been established.…”

mentioning

confidence: 99%

Evaluation of a Cytolethal Distending Toxin (cdt) Gene‐Based Species‐Specific Multiplex PCR Assay for the Identification of Campylobacter Strains Isolated from Poultry in Thailand

Samosornsuk

Asakura

Yoshida

et al. 2007

Microbiology and Immunology

View full text Add to dashboard Cite

We have recently developed a cytolethal distending toxin (cdt) gene‐based species‐specific multiplex PCR assay for identifying Campylobacter jejuni, C. coli and C. fetus. In the present study, the applicability of this assay was evaluated with 34 Campylobacter‐like organisms isolated from poultry in Thailand for species identification and was compared with other assays including API Campy, 16S rRNA gene sequence, and hippuricase (hipO) gene detection. Of the 34 strains analyzed, 20, 10 and 1 were identified as C. jejuni, C. coli, and Arcobacter cryaerophilus, respectively, and 3 could not be identified by API Campy. However, 16S rRNA gene analysis, showed that all 34 strains are C. jejuni/coli. To discriminate between these 2 species, the hipO gene, which is specifically present in C. jejuni, was examined by PCR and was detected in 20 strains, which were identified as C. jejuni by API Campy but not in the remaining 14 strains. Collective results indicated that 20 strains were C. jejuni whereas the 14 strains were C. coli. When the cdt gene‐based multiplex PCR was employed, however, 19, 20 and 19 strains were identified as C. jejuni while 13, 14 and 13 were identified as C. coli by the cdtA, cdtB and cdtC gene‐based multiplex PCR, respectively. Pulsed‐field gel electrophoresis revealed that C. jejuni and C. coli strains analyzed are genetically diverse. Taken together, these data suggest that the cdt gene‐based multiplex PCR, particularly cdtB gene‐based multiplex PCR, is a simple, rapid and reliable method for identifying the species of Campylobacter strains.

show abstract

“…Utilizou-se o multiplex-PCR proposto por Denis et al (12) adaptado por Perdoncini et al (13) para diferenciar C. jejuni e C. coli, com três pares de primers para cada reação, baseados na sequência para região 16rRNA comum entre as espécies, como segue: MD16S1 (5'-ATC TAA TGG CTT AAC CAT TAA AC -3') e MD16S2 (5' -GGA CGG TAA CTA GTT TAG TAT T -3'), com 857pb. A identificação das espécies C. jejuni e C. coli foram baseadas no gene mapA e ceuE, respectivamente, com a sequência de primers MDmapA1 (5' -CTA TTT TAT TTT TGA GTG CTT GTG -3') e MDmapA2 (5'-GCT TTA TTT GCC ATT TGT TTT ATT A -3') para o gene mapA e COL3 (5' -AAT TGA AAA TTG CTC CAA CTA TG -3') e MDCOL2 (5' -TGA TTT TAT TAT TTG TAG CAG CG-3') para o gene ceuE.…”

Section: Methodsunclassified

Campylobacter jejuni e Campylobacter coli EM CARCAÇAS DE FRANGO RESFRIADAS E CONGELADAS

Cisco

Tedesco

Perdoncini³

et al. 2017

Ciênc. anim. bras.

View full text Add to dashboard Cite

ResumoEspécies de Campylobacter spp. termotolerantes são agentes de surtos de campilobacteriose em humanos e os produtos de origem avícola são considerados uma importante fonte de infecção. Foram identificados Campylobacter jejuni e Campylobacter coli em carcaças de frango resfriadas e congeladas coletadas em três abatedouros entre 2014 e 2015. A detecção de Campylobacter spp. foi realizada por microbiologia convencional e a identificação de C. jejuni e C. coli por multiplex-PCR. Dentre as amostras avaliadas verificou-se Campylobacter spp. termotolerante em 63,8%, sendo 72,2% em carcaças resfriadas e 55,5% em carcaças congeladas. Destas, 83,3% foram positivas para C. jejuni e 66,6% para C. coli, enquanto 50% foram positivas para ambas as espécies. A presença de Campylobacter spp. termotolerante em carcaças de frangos de corte prontas para consumo representa uma importante fonte de transmissão destes patógenos para humanos. Palavras-chave: abatedouros avícolas; campilobacteriose; termotolerantes. AbstractSpecies of thermophilic Campylobacter spp. are agents of campylobacteriosis outbreaks in humans, and poultry products are implicated as major sources of infection. The detection of Campylobacter spp. was performed by conventional microbiology in poultry carcasses collected in slaughterhouses and species were identified by multiplex-PCR. Thermophilic Campylobacter spp. was verified in 63.8% of the samples, being 72.2% in chilled carcasses and 55.5% in frozen carcasses. Of these, 83.3% were positive for C. jejuni and 66.6% to C. coli, while 50% were positive for both. The presence of thermophilic Campylobacter spp. in ready-to-eat poultry products represents a potential source of human campylobacteriosis.

show abstract

Development of a m-PCR assay for simultaneous identification of Campylobacter jejuni and C. coli

Cited by 313 publications

References 16 publications

Sale of Raw Milk in Northern Italy: Food Safety Implications and Comparison of Different Analytical Methodologies for Detection of Foodborne Pathogens

Sale of Raw Milk in Northern Italy: Food Safety Implications and Comparison of Different Analytical Methodologies for Detection of Foodborne Pathogens

Evaluation of a Cytolethal Distending Toxin (cdt) Gene‐Based Species‐Specific Multiplex PCR Assay for the Identification of Campylobacter Strains Isolated from Poultry in Thailand

Campylobacter jejuni e Campylobacter coli EM CARCAÇAS DE FRANGO RESFRIADAS E CONGELADAS

Contact Info

Product

Resources

About