Publication detail

Real-time polymerase chain reaction as a tool for evaluation of magnetic poly(glycidyl methacrylate)-based microspheres in molecular diagnostics

TRACHTOVÁ, Š. ŠPANOVÁ, A. HORÁK, D. KOZÁKOVÁ, H. RITTICH, B.

Original Title

Real-time polymerase chain reaction as a tool for evaluation of magnetic poly(glycidyl methacrylate)-based microspheres in molecular diagnostics

Type

journal article in Web of Science

Language

English

Original Abstract

DNA amplification by real-time polymerase chain reaction (RT-PCR) was used for the evaluation of efficiency of polymer coating of magnetic hydrophilic poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) (P(HEMA-co-GMA)) and poly(glycidyl methacrylate) (PGMA) microspheres with/without carboxyl groups. The inhibition effect of magnetic microspheres on real-time polymerase chain reaction (RT-PCR) course was evaluated by regression analysis after the addition of different concentrations of tested microspheres to PCR mixtures. Microspheres mostly did not interfere in RT-PCR till the concentration 50 µg/25µl PCR mixture. No relationship between Fe content (and microsphere diameter) and inhibition effect was found. Microspheres containing carboxyl groups extinguished the fluorescence at lower concentrations (10-20 µg/25µl PCR mixture) without inhibition of DNA amplification as PCR products were detected using agarose gel electrophoresis. Negative effect of maghemite on PCR course was partially reduced by coating of magnetic core by silica or polymers. Two inhibition mechanisms of DNA amplification were discussed in this work.

Keywords

magnetic microspheres; inhibitory effect; real-time polymerase chain reaction; regression analysis

Authors

TRACHTOVÁ, Š.; ŠPANOVÁ, A.; HORÁK, D.; KOZÁKOVÁ, H.; RITTICH, B.

Released

27. 1. 2016

Publisher

BENTHAM SCIENCE PUBL LTD

Location

U ARAB EMIRATES

ISBN

1381-6128

Periodical

CURRENT PHARMACEUTICAL DESIGN

Year of study

22

Number

5

State

United Arab Emirates

Pages from

639

Pages to

646

Pages count

8

URL

http://www.eurekaselect.com/138077/article

BibTex

@article{BUT133353,
  author="Štěpánka {Trachtová} and Alena {Španová} and Daniel {Horák} and Hana {Kozáková} and Bohuslav {Rittich}",
  title="Real-time polymerase chain reaction as a tool for evaluation of magnetic poly(glycidyl methacrylate)-based microspheres in molecular diagnostics",
  journal="CURRENT PHARMACEUTICAL DESIGN",
  year="2016",
  volume="22",
  number="5",
  pages="639--646",
  doi="10.2174/1381612822666151228105006",
  issn="1381-6128",
  url="http://www.eurekaselect.com/138077/article"
}