Efficiency of DNA isolation methods based on silica columns and magnetic separation tested for the detection of Mycobacterium avium subsp. paratuberculosis in milk and faeces.

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F20%3AN0000139" target="_blank" >RIV/00027162:_____/20:N0000139 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216224:14310/20:00117380 RIV/62157124:16270/20:43878495

Výsledek na webu

<a href="https://www.mdpi.com/1996-1944/13/22/5112" target="_blank" >https://www.mdpi.com/1996-1944/13/22/5112</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3390/ma13225112" target="_blank" >10.3390/ma13225112</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Efficiency of DNA isolation methods based on silica columns and magnetic separation tested for the detection of Mycobacterium avium subsp. paratuberculosis in milk and faeces.

Popis výsledku v původním jazyce

Timely and reliable detection of animals shedding Mycobacterium avium subsp. paratuberculosis (MAP) should help to effectively identify infected animals and limit infection transmission at early stages to ensure effective control of paratuberculosis. The aim of the study was to compare DNA extraction methods and evaluate isolation efficiency using milk and faecal samples artificially contaminated by MAP with a focus on modern instrumental automatic DNA isolation procedures based on magnetic separation. In parallel, an automatic and manual version of magnetic separation and two methods of faecal samples preparation were compared. Commercially available DNA isolation kits were evaluated, and selected kits were used in a trial of automatic magnetic beads-based isolation and compared with the manual version of each kit. Detection of the single copy element F57 was performed by qPCR to quantify MAP and determine the isolation efficiency. The evaluated kits showed significant differences in DNA isolation efficiencies. The best results were observed with the silica column Blood and tissue kit for milk and Zymo Research for faeces. The highest isolation efficiency for magnetic separation was achieved with MagMAX for both matrices. The magnetic separation and silica column isolation methods used in this study represent frequently used methods in mycobacterial diagnostics.

Název v anglickém jazyce

Efficiency of DNA isolation methods based on silica columns and magnetic separation tested for the detection of Mycobacterium avium subsp. paratuberculosis in milk and faeces.

Popis výsledku anglicky

Timely and reliable detection of animals shedding Mycobacterium avium subsp. paratuberculosis (MAP) should help to effectively identify infected animals and limit infection transmission at early stages to ensure effective control of paratuberculosis. The aim of the study was to compare DNA extraction methods and evaluate isolation efficiency using milk and faecal samples artificially contaminated by MAP with a focus on modern instrumental automatic DNA isolation procedures based on magnetic separation. In parallel, an automatic and manual version of magnetic separation and two methods of faecal samples preparation were compared. Commercially available DNA isolation kits were evaluated, and selected kits were used in a trial of automatic magnetic beads-based isolation and compared with the manual version of each kit. Detection of the single copy element F57 was performed by qPCR to quantify MAP and determine the isolation efficiency. The evaluated kits showed significant differences in DNA isolation efficiencies. The best results were observed with the silica column Blood and tissue kit for milk and Zymo Research for faeces. The highest isolation efficiency for magnetic separation was achieved with MagMAX for both matrices. The magnetic separation and silica column isolation methods used in this study represent frequently used methods in mycobacterial diagnostics.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

40301 - Veterinary science

Projekt

<a href="/cs/project/QK1910082" target="_blank" >QK1910082: Řešení problematiky výskytu bakteriálních, protozoárních a virových zoonotických agens v chovech malých přežvýkavců</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

2020

Kód důvěrnosti údajů

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

Název periodika

Materials

ISSN

1996-1944

e-ISSN

—

Svazek periodika

13

Číslo periodika v rámci svazku

22

Stát vydavatele periodika

CH - Švýcarská konfederace

Počet stran výsledku

12

Strana od-do

"5112"

Kód UT WoS článku

000594883700001

EID výsledku v databázi Scopus

2-s2.0-85096126884