Construction of the reference standard for the detection and quantification of Mycobacterium avium subsp. paratuberculosis by quantitative PCR in faeces

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F18%3AN0000078" target="_blank" >RIV/00027162:_____/18:N0000078 - isvavai.cz</a>

Výsledek na webu

<a href="http://www.eavld2018.org/images/files/EAVLD_2018-Abstract_book.pdf" target="_blank" >http://www.eavld2018.org/images/files/EAVLD_2018-Abstract_book.pdf</a>

DOI - Digital Object Identifier

—

Jazyk výsledku

angličtina

Název v původním jazyce

Construction of the reference standard for the detection and quantification of Mycobacterium avium subsp. paratuberculosis by quantitative PCR in faeces

Popis výsledku v původním jazyce

5th Congress of the European Association of Veterinary Laboratory Diagnostics, MCE BUSINESS Conference Centre, Brussels, Belgium, 14. – 17.10.2018 – presentation. Introduction. Nowadays, quantitative PCR (qPCR) becomes a frequent method for the direct detection of Mycobacterium avium subsp. paratuberculosis (MAP). Qualitative interpretation of qPCR data is not sufficient for the determination of infectious status of the animal due to the effect of passive shedding. Therefore, precise quantification of MAP is essential. However, the quantity of MAP assessed by qPCR is dependent on the type of the qPCR quantification standard. In practise this means that quantities of MAP determined by the different qPCR assays in different laboratories are not comparable with each other. The idea of reference standard should bring the unification of the DNA standards in qPCR quantification of MAP to enable to compare quantitative data. The aim of this study was to prepare a reference quantification standard for the determination of MAP quantity in faeces by qPCR, choose the suitable storage matrix and test the stability and repeatability over the time. Material and methods. Supernatant of the faecal suspension was artificially contaminated with MAP reference strain, mixed with the matrix and lyophilized. Different types of matrices were tested. Repeatability and stability of the MAP reference standard in time were tested by qPCR. Results. We have selected the most suitable lyophilization matrix and tested stability of the reference standards in time. Discussion and Conclusion. The initial batch if reference standard is now available for the testing in routine laboratories. As OIE RL for paratuberculosis we informed OIE Biological Standards Commission to include qPCR standard among OIE-Approved Reference Standards.

Název v anglickém jazyce

Construction of the reference standard for the detection and quantification of Mycobacterium avium subsp. paratuberculosis by quantitative PCR in faeces

Popis výsledku anglicky

5th Congress of the European Association of Veterinary Laboratory Diagnostics, MCE BUSINESS Conference Centre, Brussels, Belgium, 14. – 17.10.2018 – presentation. Introduction. Nowadays, quantitative PCR (qPCR) becomes a frequent method for the direct detection of Mycobacterium avium subsp. paratuberculosis (MAP). Qualitative interpretation of qPCR data is not sufficient for the determination of infectious status of the animal due to the effect of passive shedding. Therefore, precise quantification of MAP is essential. However, the quantity of MAP assessed by qPCR is dependent on the type of the qPCR quantification standard. In practise this means that quantities of MAP determined by the different qPCR assays in different laboratories are not comparable with each other. The idea of reference standard should bring the unification of the DNA standards in qPCR quantification of MAP to enable to compare quantitative data. The aim of this study was to prepare a reference quantification standard for the determination of MAP quantity in faeces by qPCR, choose the suitable storage matrix and test the stability and repeatability over the time. Material and methods. Supernatant of the faecal suspension was artificially contaminated with MAP reference strain, mixed with the matrix and lyophilized. Different types of matrices were tested. Repeatability and stability of the MAP reference standard in time were tested by qPCR. Results. We have selected the most suitable lyophilization matrix and tested stability of the reference standards in time. Discussion and Conclusion. The initial batch if reference standard is now available for the testing in routine laboratories. As OIE RL for paratuberculosis we informed OIE Biological Standards Commission to include qPCR standard among OIE-Approved Reference Standards.

Druh

O - Ostatní výsledky

CEP obor

—

OECD FORD obor

10606 - Microbiology

Projekt

<a href="/cs/project/VI20152020044" target="_blank" >VI20152020044: Multiplexní xMAP technologie pro komplexní detekci patogenních agens významných z pohledu zajištění ochrany zdraví lidí a zvířat</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2018

Kód důvěrnosti údajů

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