Detekce Mycobacterium avium subsp. paratuberculosis v nepasterovaném mléce pomocí IS900 and F57 kompetitivní real time kvantitativní PCR a kultivace

Kód výsledku v IS VaVaI

RIV/00027162:_____/08:#0000429 - isvavai.cz

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

On-farm spread of Mycobacterium avium subsp. paratuberculosis in raw milk studied by IS900 and F57 competitive real time quantitative PCR and culture examination

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

On-farm spread of Mycobacterium avium subsp. paratuberculosis in raw milk studied by IS900 and F57 competitive real time quantitative PCR and culture examination

Popis výsledku anglicky

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.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

GJ - Choroby a škůdci zvířat, veterinární medicina

OECD FORD obor

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Projekt

QH81065: Tlumení paratuberkulózy v České republice: zavedení nových metod pro urychlení detekce původce, sledování jeho přežívání a šíření v chovech, jeho distribuce v prostředí a posouzení rizik kontaminace krmiv a potravin

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2008

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

International Journal of Food Microbiology

ISSN

0168-1605

e-ISSN

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Svazek periodika

128

Číslo periodika v rámci svazku

2

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

8

Strana od-do

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Kód UT WoS článku

000261668100010

EID výsledku v databázi Scopus

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