| dc.contributor.author |
Rodriguez-Lazaro, D |
en |
| dc.contributor.author |
Lloyd, J |
en |
| dc.contributor.author |
Herrewegh, A |
en |
| dc.contributor.author |
Ikonomopoulos, J |
en |
| dc.contributor.author |
D'Agostino, M |
en |
| dc.contributor.author |
Pla, M |
en |
| dc.contributor.author |
Cook, N |
en |
| dc.date.accessioned |
2014-06-06T06:45:53Z |
|
| dc.date.available |
2014-06-06T06:45:53Z |
|
| dc.date.issued |
2004 |
en |
| dc.identifier.issn |
03781097 |
en |
| dc.identifier.uri |
http://dx.doi.org/10.1016/j.femsle.2004.06.024 |
en |
| dc.identifier.uri |
http://62.217.125.90/xmlui/handle/123456789/2688 |
|
| dc.subject |
Food products |
en |
| dc.subject |
Internal amplification control (IAC) |
en |
| dc.subject |
Milk |
en |
| dc.subject |
Mycobacterium avium subsp. paratuberculosis |
en |
| dc.subject |
Real-time NASBA |
en |
| dc.subject.other |
water |
en |
| dc.subject.other |
addition reaction |
en |
| dc.subject.other |
article |
en |
| dc.subject.other |
bacterium detection |
en |
| dc.subject.other |
bacterium identification |
en |
| dc.subject.other |
centrifugation |
en |
| dc.subject.other |
controlled study |
en |
| dc.subject.other |
diagnostic accuracy |
en |
| dc.subject.other |
extraction |
en |
| dc.subject.other |
gene amplification |
en |
| dc.subject.other |
gene identification |
en |
| dc.subject.other |
gene targeting |
en |
| dc.subject.other |
milk |
en |
| dc.subject.other |
molecular dynamics |
en |
| dc.subject.other |
Mycobacterium avium |
en |
| dc.subject.other |
Mycobacterium paratuberculosis |
en |
| dc.subject.other |
nonhuman |
en |
| dc.subject.other |
nucleic acid analysis |
en |
| dc.subject.other |
priority journal |
en |
| dc.subject.other |
Animals |
en |
| dc.subject.other |
Bacterial Proteins |
en |
| dc.subject.other |
DNA, Bacterial |
en |
| dc.subject.other |
DNA-Binding Proteins |
en |
| dc.subject.other |
Food Microbiology |
en |
| dc.subject.other |
Fresh Water |
en |
| dc.subject.other |
Genes, Bacterial |
en |
| dc.subject.other |
Milk |
en |
| dc.subject.other |
Mycobacterium paratuberculosis |
en |
| dc.subject.other |
Quality Control |
en |
| dc.subject.other |
Self-Sustained Sequence Replication |
en |
| dc.subject.other |
Sensitivity and Specificity |
en |
| dc.subject.other |
Water Microbiology |
en |
| dc.subject.other |
Bacteria (microorganisms) |
en |
| dc.subject.other |
Mycobacterium |
en |
| dc.subject.other |
Mycobacterium avium |
en |
| dc.subject.other |
Mycobacterium avium subsp. paratuberculosis |
en |
| dc.title |
A molecular beacon-based real-time NASBA assay for detection of Mycobacterium avium subsp. paratuberculosis in water and milk |
en |
| heal.type |
journalArticle |
en |
| heal.identifier.primary |
10.1016/j.femsle.2004.06.024 |
en |
| heal.publicationDate |
2004 |
en |
| heal.abstract |
A molecular beacon-based real-time NASBA assay for detection and identification of Mycobacterium avium subsp. paratuberculosis has been developed. It targets and amplifies sequences from the dnaA gene which are specific for this bacterium. The assay includes an internal amplification control, to allow identification of inhibited reactions. The assay was tested against 18 isolates of M. avium subsp. paratuberculosis, 17 other mycobacterial strains and 25 non-mycobacterial strains, and was fully selective in that it detected all the targets but none of the non-targets. The lowest number of cells which the assay can detect with 99% probability is 150-200 cells per reaction (as determined using pure culture suspensions). Using centrifugation and nucleic acid extraction as sample treatment, the assay was able to consistently detect 103 M. avium subsp. paratuberculosis cells in 20 ml 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 104 M. avium subsp. paratuberculosis cells 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 M. avium subsp. paratuberculosis. Crown Copyright © 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved. |
en |
| heal.journalName |
FEMS Microbiology Letters |
en |
| dc.identifier.issue |
1 |
en |
| dc.identifier.volume |
237 |
en |
| dc.identifier.doi |
10.1016/j.femsle.2004.06.024 |
en |
| dc.identifier.spage |
119 |
en |
| dc.identifier.epage |
126 |
en |