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Isolation and PCR amplification of a species-specific oxidoreductase-coding gene region in Listeria grayi

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Abstract:

Listeria grayi is a nonpathogenic Gram-positive bacterium that demonstrates considerable similarities to other members in the genus Listeria, including the foodborne human pathogen Listeria monocytogenes and the animal pathogen Listeria ivanovii. A rapid diagnostic test to identify and diagnose listeriosis would be valuable, especially in cases where the presence of L. grayi may complicate diagnosis. This test would be based on a unique gene present in L. grayi. In this study, after comparative screening of a recombinant L. grayi DNA library by dot blot hybridization, an L. grayi specific clone (lgr20-246) with an insert of 722 bp was isolated. By applying PCR primers derived from a distinct region of the clone not shared by other bacteria, a specific band of 420 bp was amplified from the genomic DNA of L. grayi only and not of other Listeria species or common bacteria. These results suggest that the PCR assay employing primers lgr20-246F and lgr20-246R provides an independent and precise means of distinguishing L. grayi from other Listeria species and common bacteria. Therefore, it would be another useful technique for laboratory differentiation of Listeria bacteria.Key words: Listeria grayi, PCR, identification, diagnosis.

Listeria grayi est une bactérie Gram-positive non pathogène qui démontre de grandes similitudes avec d'autres membres du genre Listeria, incluant la pathogène alimentaire humain Listeria monocytogenes et la pathogène animal Listeria ivanovii. La disponibilité d'un test de diagnostic rapide basé sur un gène présent uniquement chez L. grayi serait utile lorsque la présence de L. grayi pourrait compliquer l'identification et le diagnostic de la listériose. Dans cette étude, un clone spécifique de L. grayi (Igr20-246) renfermant un insert de 722 pb a été isolé suite à un criblage comparatif d'une banque d'ADN recombinant de L. grayi par hybridation sur transfert en point. En employant des amorces de PCR issues d'une région distincte du clone qui n'est pas partagée avec les autres bactéries, une bande spécifique de 420 pb a été amplifiée à partir de l'ADN génomique de L. grayi seulement, mais non d'autres Listeria ou d'autres bactéries communes. Ces résultats indiquent que le test de PCR employant les amorces lgr20-246F et lgr20-246R fournit un moyen indépendant et précis de distinguer L. grayi d'autres espèces de Listeria ou d'autres bactéries communes, et représente un ajout utile aux techniques existantes pour la différenciation des bactéries Listeria en laboratoire.Mots clés : Listeria grayi, PCR, identification, diagnostic.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: 2005-01-01

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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