Comparison of filtering methods, filter processing and DNA extraction kits for detection of mycobacteria in water

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F15%3AN0000008" target="_blank" >RIV/00027162:_____/15:N0000008 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00027162:_____/15:#0001245

Výsledek na webu

<a href="http://www.aaem.pl/Comparison-of-filtering-methods-filter-processing-and-DNA-extraction-kits-for-detection-of-mycobacteria-in-water,72303,0,2.html" target="_blank" >http://www.aaem.pl/Comparison-of-filtering-methods-filter-processing-and-DNA-extraction-kits-for-detection-of-mycobacteria-in-water,72303,0,2.html</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.5604/12321966.1167707" target="_blank" >10.5604/12321966.1167707</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Comparison of filtering methods, filter processing and DNA extraction kits for detection of mycobacteria in water

Popis výsledku v původním jazyce

Introduction and objective. Mycobacteria have been isolated from almost all types of natural waters, as well as from man-made water distribution systems. Detection of mycobacteria using PCR has been described in different types of water; however, currently, there is no standardised protocol for the processing of large volumes of water. Material and Methods. In the present study, different filtering methods are tested and optimised for tap or river water filtration up to 10 L, as well as filter processing and DNA isolation using four commercially available kits. Results. The PowerWater DNA isolation kit (MoBio, USA), together with a kit used for soil and other environmental samples (PowerSoil DNA isolation kit, MoBio), had the highest efficiency. Filtration of 10 L of water and elution of the filter in PBS with the addition of 0.05% of Tween 80 is suggested. Conclusions. The described protocol for filter elution is recommended, and the use of the PowerWater DNA isolation kit for the highest mycobacterial DNA yield from water samples. The described protocol is suitable for parallel detection of mycobacteria using cultivation.

Název v anglickém jazyce

Comparison of filtering methods, filter processing and DNA extraction kits for detection of mycobacteria in water

Popis výsledku anglicky

Introduction and objective. Mycobacteria have been isolated from almost all types of natural waters, as well as from man-made water distribution systems. Detection of mycobacteria using PCR has been described in different types of water; however, currently, there is no standardised protocol for the processing of large volumes of water. Material and Methods. In the present study, different filtering methods are tested and optimised for tap or river water filtration up to 10 L, as well as filter processing and DNA isolation using four commercially available kits. Results. The PowerWater DNA isolation kit (MoBio, USA), together with a kit used for soil and other environmental samples (PowerSoil DNA isolation kit, MoBio), had the highest efficiency. Filtration of 10 L of water and elution of the filter in PBS with the addition of 0.05% of Tween 80 is suggested. Conclusions. The described protocol for filter elution is recommended, and the use of the PowerWater DNA isolation kit for the highest mycobacterial DNA yield from water samples. The described protocol is suitable for parallel detection of mycobacteria using cultivation.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10606 - Microbiology

Projekt

<a href="/cs/project/LO1218" target="_blank" >LO1218: Zdravé zvíře jako zdroj zdravé potraviny</a><br>

Návaznosti

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

Rok uplatnění

2015

Kód důvěrnosti údajů

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

Název periodika

ANNALS OF AGRICULTURAL AND ENVIRONMENTAL MEDICINE

ISSN

1232-1966

e-ISSN

1898-2263

Svazek periodika

22

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

PL - Polská republika

Počet stran výsledku

4

Strana od-do

429-432

Kód UT WoS článku

000364164500008

EID výsledku v databázi Scopus

2-s2.0-84941617248