Use of a single procedure for selective enrichment, isolation, and identification of plasmid-bearing virulent Yersinia enterocolitica of various serotypes from pork samples

Bhaduri, Saumya; Cottrell, Bryan J.; Pickard, Allan R.

doi:10.1128/aem.63.5.1657-1660.1997

Cited by 42 publications

(26 citation statements)

References 21 publications

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“…Sublethally damaged cells may fail to grow during the enrichment step and are thus less likely to be detected by a method involving enrichment than by direct detection. Application of a non‐selective enrichment medium will diminish this problem by avoiding the selective substances that suppress growth, especially during the initial lag phase ( Bhaduri et al . 1997 ).…”

Section: Discussionmentioning

confidence: 99%

Detection of low numbers of pathogenic Yersinia enterocolitica in environmental water and sewage samples by nested polymerase chain reaction

Waage

Vardund

Lund³

et al. 1999

J Appl Microbiol

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A.S. WAAGE, T. VARDUND, V. LUND and G. KAPPERUD.1999.Isolation of pathogenic Yersinia enterocolitica from water and sewage by traditional culture techniques is time‐consuming and subsequent differentiation between pathogenic and non‐pathogenic strains can be difficult and unreliable. A nested polymerase chain reaction (PCR) procedure was used for the detection of low numbers of Y. enterocolitica in spiked samples from natural surface sources with variable background flora ranging from oligotrophic water to sewage. Water and sewage samples were filtered and filters enriched overnight in a non‐selective medium. Nested PCR conducted on enriched broth, prepared by use of a rapid and simple preparation step consisting of centrifugation, proteinase K treatment and boiling, enabled the detection of 8–17 cfu 100 ml−1 water with background levels of up to 8700 heterotrophic organisms ml−1 and 10 000 cfu coliform organisms 100 ml−1 water. The analysis can be completed within 2–3 d and should be a significant tool in monitoring environmental waters and drinking water sources for the presence of pathogenic Y. enterocolitica.

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Section: Discussionmentioning

confidence: 99%

Detection of low numbers of pathogenic Yersinia enterocolitica in environmental water and sewage samples by nested polymerase chain reaction

Waage

Vardund

Lund³

et al. 1999

J Appl Microbiol

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“…Isolation and identi®cation of Y. enterocolitica and Y. pseudotuberculosis strains was carried out by the method of Bhaduri et al (1997) modi®ed by us. Brie¯y, small 10-g samples of organs were placed in 50 ml tryptic soy broth (TSB) supplemented with 0.3% sodium deoxycholate, 4 mg/ml irgasan and 0.01 ml Tween-80.…”

Section: Isolation and Identi®cationmentioning

confidence: 99%

Isolation of Pathogenic Yersiniae from Wild Animals in Bulgaria

Николова

Tzvetkov²,

Najdenski

et al. 2001

Journal of Veterinary Medicine, Series B

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Pathogenic Yersinia strains were isolated between December 1998 and April 1999 from 37 wild animals: rabbit (Lepus europeus), boar (Sus scrofa scrofa), asiatic jackal (Canis aureus), red fox (Vulpes vulpes), mouflon (Ovis musimon), european river otter (Lutra lutra), beech marten (Martes foina), polecat (Musleta putorius) and wild cat (Felis silvestris). It was established that among the wild animals Y. enterocolitica strains of serotype 0:3 predominated, accompanied by Y. pseudotuberculosis strains of serotype 0:3. In one sample from asiatic jackal and one sample from rabbit, Y. enterocolitica serotype 0:8 was isolated. Yersinia enterocolitica and Y. pseudotuberculosis strains were isolated from tonsils and tongues as well as from the viscera--lung, liver, heart, spleen, kidney and lymph nodes, mainly in young animals (1-2 years of age). The results showed that wild animals are a possible natural reservoir for pathogenic Y. enterocolitica and Y. pseudotuberculosis and are included in the epidemiological chain of yersinioses.

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“…Ground pork (n = 106) samples were obtained from local market and processed for selective enrichment based on the method of Bhaduri et al (1997). Briefly, 10 g of ground pork was aseptically mixed in 90 ml of Yersinia enrichment broth.…”

Section: Isolation Of Y Enterocolitica From Pork Samplesmentioning

confidence: 99%

Cloning, expression and characterization of attachment-invasion locus protein (Ail) ofYersinia enterocoliticaand its utilization in rapid detection by immunoassays

Balakrishna

Murali

Batra

2010

Letters in Applied Microbiology

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Aims: Rapid detection of pathogenic Yersinia enterocolitica isolates by using antisera raised against recombinant attachment‐invasion locus (Ail) protein. Methods and Results: The complete gene (471 bp) encoding for the Ail protein was amplified by PCR and cloned in pQE 30 UA vector. The recombinant clones were selected by polymerase chain reaction (PCR). Recombinant protein was expressed using induction with 1 mmol l−1 final concentration of isopropylthiogalactoside (IPTG). Polyclonal antibodies were raised in mice against this purified recombinant protein. An indirect plate ELISA was standardized based on rAil protein for the detection of Y. enterocolitica. Western blot analysis with the sera raised against recombinant Ail protein exhibited reaction at 17 kDa region of the native Ail protein present in pathogenic Y. enterocolitica standard strains and strains isolated from pork samples suggesting that the antigenicity of recombinant Ail protein was similar to that of native Ail protein. Nonpathogenic Y. enterocolitica and the other species of Yersinia, namely, Y. pseudotuberculosis, Y. intermedia, Y. kristenseni, Y. fredrickseni and also the Enterobacteriaceae organisms tested were not found reacting to polyclonal antisera against this recombinant Ail protein. Conclusion: The antibodies raised against recombinant Ail protein could specifically identify pathogenic Y. enterocolitica strains both by indirect plate ELISA and Western blot immunoassay. Significance and Impact of the Study: The method developed in this study may find application in the detection of pathogenic Y. enterocolitica not only from food and environmental samples but also from clinical samples.

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Use of a single procedure for selective enrichment, isolation, and identification of plasmid-bearing virulent Yersinia enterocolitica of various serotypes from pork samples

Cited by 42 publications

References 21 publications

Detection of low numbers of pathogenic Yersinia enterocolitica in environmental water and sewage samples by nested polymerase chain reaction

Detection of low numbers of pathogenic Yersinia enterocolitica in environmental water and sewage samples by nested polymerase chain reaction

Isolation of Pathogenic Yersiniae from Wild Animals in Bulgaria

Cloning, expression and characterization of attachment-invasion locus protein (Ail) ofYersinia enterocoliticaand its utilization in rapid detection by immunoassays

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