Comparison of hipO and ceuE Gene Based PCR Assays for the Detection of Campylobacter Jejuni

Ayaz, ND

doi:10.17352/jcmbt.000008

Cited by 4 publications

(3 citation statements)

References 20 publications

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“…The choice of method will depend on the relevant conditions. Some authors use the boiling method (Shams and others ; Zang and others ), others prefer the kit (Frasao and others , b; Ayaz and others ; Gosselin‐Theberge and others ; Rodgers and others ), whereas others employ phenol‐chloroform‐isoamyl alcohol (25: 24: 1) extraction (Raja and others ).…”

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

confidence: 99%

“…However, quantification is excluded (Postollec and others ). With PCR, specificity is enhanced by using primers specific for Campylobacter (von Altrock and others ; Lemos and others ; Han and others ), C. coli (Gonzalez and others ; von Altrock and others ; Fontanot and others ; Frasao and others , b; Lemos and others ; Han and others ), C. jejuni (von Altrock and others ; Fontanot and others ; Lemos and others ; Ayaz and others ; Han and others ; Raja and others ), C. lari (Fontanot and others ), and C. upsaliensis (Fontanot and others ) (Table ). In addition to these specific primers, it is possible to amplify to different sizes (bp) by using a primer linked to the 16S‐23S ITS sequence, as this region has a variable size and is interspersed within well‐conserved regions, allowing differentiation between C. jejuni and C. coli (Fontanot and others ).…”

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

confidence: 99%

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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%

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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“…In the last ten years, the occurrence of Campylobacteriosis were in an increasing trend. As C. jejuni able to colonize poultry intestine that lead to poultry meat a reservoir of foodborne Campylobacteriosis (Ayaz et al, 2016). The main route of transmission of infection is eating of food of animal origin and particularly poultry meat (Butzler and Oosterom, 1991).…”

Section: Discussionmentioning

confidence: 99%

Phenotypic and genotypic chactracterization of Campylobacter isolated from poultry

Ahmed

El-Tawab

Hotzel

et al. 2019

Benha Veterinary Medical Journal

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Campylobacteriosis incidence and proportion of Campylobacter strains resistant to antibiotic have been increased worldwide in the last decades. Campylobacteriosis is considered as one of the major important zoonotic gastrointestinal diseases around the world. The aim of this study was isolation and biochemical characterization of Campylobacter from poultry. Campylobacter isolation have been done on MCCDA media, motility has been detected by phase contrast microscope, morphology has been detected by Gram stain, and confirmed by MALDI-TOF MS and molecular confirmation by using cPCR by amplification of 16S rRNA gene. A total of 102 isolates of Campylobacter were isolated from farm and small backyards located in different sources in El-Kalyobia , El-Monofia and El-sharkia Governorates by conventional bacteriological methods from which 40 were C. jejuni and 62 were C. coli. All isolates have been confirmed as Campylobacter by MALDI-TOF MS and detection of 16S rRNA by cPCR. PCR is a useful molecular tool for identification and confirmation of Campylobacter. It is rapid, sensitive, and specific than the culture methods, but the only disadvantage is that is expensive.

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Antibody-conjugated ferromagnetic nanoparticles with lateral flow test strip assay for rapid detection of Campylobacter jejuni in poultry samples

Poonlapdecha

Seetang-Nun

Wonglumsom

et al. 2018

International Journal of Food Microbiology

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The aim of this study was to develop a nanoparticle-based cell capture system combined with a lateral flow test strip (LFT) assay for rapid detection of Campylobacter jejuni from poultry samples. The developed assay was bench-marked against the standard modified Charcoal Cefoperazone Deoxycholate Agar (mCCDA) method according to ISO16140:2003 procedures. The synthesized ferromagnetic nanoparticles (FMNs) were modified with glutaraldehyde, then functionalized with polyclonal antibodies for specific C. jejuni capture and concentration from poultry samples. After lysing captured cells, DNA from C. jejuni was amplified by PCR using the primers designed to target the hipO gene, and the PCR amplicons were detected with the lateral flow test strip assay. Following the ISO16140:2003 guidelines, the relative detection limit, and the inclusivity and exclusivity tests were determined. The results showed that the limit of detection (LOD) of the assay was 10 or 1 cfu/ml with C. jejuni in pure culture and 10-10 cfu/ml with target cells spiked in poultry sample. In addition, the inclusivity and exclusivity tests were found to be 100%. Using field chicken samples (n = 60), the assay showed relative accuracy, relative specificity, and relative sensitivity of 96.67%, 100% and 93.33%, respectively. The positive predictive values (PPV) and negative predictive values (NPV), and the kappa index of concordance (k) were calculated as 100% and 93.75%, and 0.93, respectively. The developed assay required approximately 3 h to complete and gave results comparable to those analyzed by the standard culture method, which required 5-7 days. The assay is rapid, easy-to-use, and has potential to be directly applied to C. jejuni detection in various categories of poultry samples.

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Comparison of hipO and ceuE Gene Based PCR Assays for the Detection of Campylobacter Jejuni

Cited by 4 publications

References 20 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

Phenotypic and genotypic chactracterization of Campylobacter isolated from poultry

Antibody-conjugated ferromagnetic nanoparticles with lateral flow test strip assay for rapid detection of Campylobacter jejuni in poultry samples

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