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Real-time PCR-based methods for detection of Mycobacterium avium Subsp paratuberculosis in water and milk

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dc.contributor.author Rodriguez-Lazaro, D en
dc.contributor.author D'Agostino, M en
dc.contributor.author Herrewegh, A en
dc.contributor.author Pla, M en
dc.contributor.author Cook, N en
dc.contributor.author Ikonomopoulos, J en
dc.date.accessioned 2014-06-06T06:46:44Z
dc.date.available 2014-06-06T06:46:44Z
dc.date.issued 2005 en
dc.identifier.issn 0168-1605 en
dc.identifier.uri http://62.217.125.90/xmlui/handle/123456789/3167
dc.subject Johne's disease en
dc.subject Crohn's disease en
dc.subject real-time PCR en
dc.subject pathogen detection en
dc.subject milk en
dc.subject water en
dc.subject.classification Food Science & Technology en
dc.subject.classification Microbiology en
dc.subject.other CROHNS-DISEASE en
dc.subject.other QUANTITATIVE PCR en
dc.subject.other DIAGNOSTIC PCR en
dc.subject.other LISTERIA-MONOCYTOGENES en
dc.subject.other SALMONELLA-ENTERICA en
dc.subject.other HEAT-RESISTANCE en
dc.subject.other JOHNES-DISEASE en
dc.subject.other FOOD SAMPLES en
dc.subject.other IDENTIFICATION en
dc.subject.other AMPLIFICATION en
dc.title Real-time PCR-based methods for detection of Mycobacterium avium Subsp paratuberculosis in water and milk en
heal.type journalArticle en
heal.language English en
heal.publicationDate 2005 en
heal.abstract A real-time PCR assay for quantitative detection of Mycobacterium avium paratuberculosis has been developed. It targets and amplifies sequences from the IS900 insertion element which is specific for this bacterium, and includes an internal amplification control. The assay was tested against 18 isolates of M. avium paratuberculosis, 17 other mycobacterial strains, and 25 non-mycobacterial strains, and was fully selective. It is capable of detecting < 3 genomic DNA copies with 99% probability or alternatively, using cells directly in the reaction, 12 cells can be detected with 99% probability. Using prior centrifugation, the assay was able to consistently and quantifiably detect 10(2) M. avium paratuberculosis cells in 20 all artificially contaminated drinking water. With a simple detergent and enzymatic sample pretreatment before centrifugation and nucleic acid extraction, the assay was able to consistently detect 10(2) M. avium paratuberculosis in 20 ml artificially contaminated semi-skimmed milk. The assay will be a useful addition to the range of diagnostic tools available for the study of All. avium paratuberculosis. Crown Copyright © 2004 Elsevier B.V. All rights reserved. en
heal.publisher ELSEVIER SCIENCE BV en
heal.journalName INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY en
dc.identifier.issue 1 en
dc.identifier.volume 101 en
dc.identifier.isi ISI:000229157300010 en
dc.identifier.spage 93 en
dc.identifier.epage 104 en


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