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A New Sensitive Method for the Detection of Mycoplasmas Using Fluorescence Microscopy

The result's identifiers

  • Result code in IS VaVaI

    <a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15110%2F19%3A73596065" target="_blank" >RIV/61989592:15110/19:73596065 - isvavai.cz</a>

  • Alternative codes found

    RIV/00216275:25310/19:39917055

  • Result on the web

    <a href="https://www.mdpi.com/2073-4409/8/12/1510/htm" target="_blank" >https://www.mdpi.com/2073-4409/8/12/1510/htm</a>

  • DOI - Digital Object Identifier

    <a href="http://dx.doi.org/10.3390/cells8121510" target="_blank" >10.3390/cells8121510</a>

Alternative languages

  • Result language

    angličtina

  • Original language name

    A New Sensitive Method for the Detection of Mycoplasmas Using Fluorescence Microscopy

  • Original language description

    Contamination of cell cultures by mycoplasmas is a very common phenomenon. As they can substantially alter cell metabolism and potentially spread to all cell cultures in laboratory, their early detection is necessary. One of the fastest and cheapest methods of mycoplasma detection relies on the direct staining of mycoplasmas’ DNA by DAPI or Hoechst dyes. Although this method is easy and fast to perform, it suffers from the low signal provided by these dyes compared to the nuclear DNA. Therefore, the reporter cell lines are used for cultivation of mycoplasmas before DAPI or the Hoechst staining step. In the study presented, we have developed and tested a new immunofluorescence assay for the detection of mycoplasmas. The method is based on the enzymatic labeling using DNA polymerase I and modified nucleotides utilizing nicks in the mycoplasmas’ DNA. Modified nucleotides are incorporated into mycoplasmas’ DNA and subsequently visualized by immunofluorescence microscopy. The developed approach is independent of the mycoplasma strain, does not intensely stain nuclear DNA, does not stain other bacteria, and provides higher sensitivity than the approach based on the direct labeling using DAPI or Hoechst dyes.

  • Czech name

  • Czech description

Classification

  • Type

    J<sub>ost</sub> - Miscellaneous article in a specialist periodical

  • CEP classification

  • OECD FORD branch

    10606 - Microbiology

Result continuities

  • Project

    Result was created during the realization of more than one project. More information in the Projects tab.

  • Continuities

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

Others

  • Publication year

    2019

  • Confidentiality

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

Data specific for result type

  • Name of the periodical

    Cells

  • ISSN

    2073-4409

  • e-ISSN

  • Volume of the periodical

    2019

  • Issue of the periodical within the volume

    8(12):1510

  • Country of publishing house

    CH - SWITZERLAND

  • Number of pages

    17

  • Pages from-to

  • UT code for WoS article

    000506645100002

  • EID of the result in the Scopus database