On-farm spread of Mycobacterium avium subsp. paratuberculosis in raw milk studied by IS900 and F57 competitive real time quantitative PCR and culture examination
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F08%3A%230000429" target="_blank" >RIV/00027162:_____/08:#0000429 - isvavai.cz</a>
Result on the web
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DOI - Digital Object Identifier
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Alternative languages
Result language
angličtina
Original language name
On-farm spread of Mycobacterium avium subsp. paratuberculosis in raw milk studied by IS900 and F57 competitive real time quantitative PCR and culture examination
Original language description
We developed two duplex real time qPCR systems specific for Mycobacterium avium subsp. paratuberculosis (MAP) detection. These real time qPCR assays amplify the multicopy element IS900 for qualitative analysis and the single copy element F57 for quantitative analysis. Both assays incorporate an internal amplification control. The sensitivity of the developed assays and isolation efficiency were demonstrated through the analysis of raw milk samples artificially contaminated with MAP cells and with plasmids containing cloned fragments of the targets. The developed assays for milk analysis were applied to samples from one farm with two faecal shedding cows. Three hundred and forty five individual milk samples were tested by real time qPCR assays and by cultivation. Hundred and eleven (32.5%) individual milk samples were positive by the real time qPCR, no milk sample was culture positive. The spread of MAP in individual, tank and bulk tank milk samples was also monitored.
Czech name
Detekce Mycobacterium avium subsp. paratuberculosis v nepasterovaném mléce pomocí IS900 and F57 kompetitivní real time kvantitativní PCR a kultivace
Czech description
Cílem studie bylo vyvinout duplexní real time qPCR systémy s interní amplifikační kontrolou specifické pro detekci Mycobacterium avium subsp. paratuberculosis (MAP). Jako detekční cíle byly zvoleny pro kvalitativní analýzu multikopiový IS900 element a pro kvantifikační analýzu jednokopiový F57 element. Citlivost vyvinutých PCR systému a účinnost izolace byly stanoveny pomocí umělé kontaminace nepasterovaného mléka definovanou bakteriální suspenzí a zároveň pomocí plazmidových konstruktů. Vyvinuté systémy byly použity na analýzu.
Classification
Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
GJ - Diseases and animal vermin, veterinary medicine
OECD FORD branch
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Result continuities
Project
<a href="/en/project/QH81065" target="_blank" >QH81065: Suppression of paratuberculosis in the CZ: introduction of new methods for the causative agent detection, monitoring of its survival and spread in studs, its distribution in the environment and examination of food and feed contamination.</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>Z - Vyzkumny zamer (s odkazem do CEZ)
Others
Publication year
2008
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
International Journal of Food Microbiology
ISSN
0168-1605
e-ISSN
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Volume of the periodical
128
Issue of the periodical within the volume
2
Country of publishing house
NL - THE KINGDOM OF THE NETHERLANDS
Number of pages
8
Pages from-to
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UT code for WoS article
000261668100010
EID of the result in the Scopus database
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