Comparative analysis of yadA and ail polymerase chain reaction methods for virulent Yersinia enterocolitica

Blais, Burton W.; Phillippe, Lucille M.

doi:10.1016/0956-7135(95)00015-j

Cited by 32 publications

(16 citation statements)

References 11 publications

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“…When MALDI-TOF MS did not provide sufficient interspecies discrimination, identification was performed by colony-PCR amplification and sequencing of the 16S rRNA genes using the fD1/rp2 primers designed by Weisburg et al (17). All clinical isolates of Y. enterocolitica were screened for the yadA gene, using the PCR assay described by Blais and Phillippe (18).…”

Section: Methodsmentioning

confidence: 99%

CHROMagar Yersinia, a New Chromogenic Agar for Screening of Potentially Pathogenic Yersinia enterocolitica Isolates in Stools

et al. 2013

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CHROMagar Yersinia (CAY) is a new chromogenic medium for the presumptive detection of virulent Yersinia enterocolitica in stools. Based on a comparative analysis of 1,494 consecutive stools from hospitalized patients, CAY was found to be just as sensitive as the reference medium (cefsulodin-irgasan-novobiocin agar) but was significantly more specific and had a very low falsepositive rate. CAY reduces the workload (and thus costs) for stool analysis and can therefore be recommended for routine laboratory use.

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Section: Methodsmentioning

confidence: 99%

CHROMagar Yersinia, a New Chromogenic Agar for Screening of Potentially Pathogenic Yersinia enterocolitica Isolates in Stools

et al. 2013

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“…In contrast, apathogenic strains of Y. enterocolitica do not contain these two genes. However, the plasmid harboring the yadA gene can be lost under certain cultivation conditions in the laboratory (4). This may lead to false-negative results in any test system based on the presence of this plasmid.…”

mentioning

confidence: 95%

Identification ofYersinia enterocoliticaat the Species and Subspecies Levels by Fourier Transform Infrared Spectroscopy

Kuhm

Suter

Felleisen

et al. 2009

Appl Environ Microbiol

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intermedia, and Y. rohdei strains from Y. enterocolitica, which could not be differentiated by the API 20E test system. The probability for correct biotype identification of Y. enterocolitica isolates was 98.3% (41 externally validated strains). For correct serotype identification, the probability was 92.5% (42 externally validated strains). In addition, the presence or absence of the ail gene, one of the main pathogenicity markers, was demonstrated using FT-IR. The probability for correct identification of isolates concerning the ail gene was 98.5% (51 externally validated strains). This indicates that it is possible to obtain information about genus, species, and in the case of Y. enterocolitica also subspecies type with a single measurement. Furthermore, this is the first example of the identification of specific pathogenicity using FT-IR.

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“…Virulence in Y. enterocolitica results from a complex interplay between a series of plasmid-and chromosomally located genes. However, PCR targets located on the virulence plasmid must be considered unsuitable as targets for detection because the plasmid is unstable and easily lost during laboratory treatment (5,22).…”

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confidence: 99%

Real-Time PCR Method for Detection of Pathogenic Yersinia enterocolitica in Food

Lambertz

Nilsson

Hallanvuo³

et al. 2008

Appl Environ Microbiol

116

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The current methods for the detection of pathogenic Yersinia enterocolitica bacteria in food are time consuming and inefficient. Therefore, we have developed and evaluated in-house a TaqMan probe-based real-time PCR method for the detection of this pathogen. The complete method comprises overnight enrichment, DNA extraction, and real-time PCR amplification. Also included in the method is an internal amplification control. The selected primer-probe set was designed to use a 163-bp amplicon from the chromosomally located gene ail (attachment and invasion locus). The selectivity of the PCR method was tested with a diverse range (n ‫؍‬ 152) of related and unrelated strains, and no false-negative or false-positive PCR results were obtained. The sensitivity of the PCR amplification was 85 fg purified genomic DNA, equivalent to 10 cells per PCR tube. Following the enrichment of 10 g of various food samples (milk, minced beef, cold-smoked sausage, fish, and carrots), the sensitivity ranged from 0.5 to 55 CFU Y. enterocolitica. Good precision, robustness, and efficiency of the PCR amplification were also established. In addition, the method was tested on naturally contaminated food; in all, 18 out of 125 samples were positive for the ail gene. Since no conventional culture method could be used as a reference method, the PCR products amplified from these samples were positively verified by using conventional PCR and sequencing of the amplicons. A rapid and specific real-time PCR method for the detection of pathogenic Y. enterocolitica bacteria in food, as presented here, provides a superior alternative to the currently available detection methods and makes it possible to identify the foods at risk for Y. enterocolitica contamination.

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Comparative analysis of yadA and ail polymerase chain reaction methods for virulent Yersinia enterocolitica

Cited by 32 publications

References 11 publications

CHROMagar Yersinia, a New Chromogenic Agar for Screening of Potentially Pathogenic Yersinia enterocolitica Isolates in Stools

CHROMagar Yersinia, a New Chromogenic Agar for Screening of Potentially Pathogenic Yersinia enterocolitica Isolates in Stools

Identification ofYersinia enterocoliticaat the Species and Subspecies Levels by Fourier Transform Infrared Spectroscopy

Real-Time PCR Method for Detection of Pathogenic Yersinia enterocolitica in Food

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