Toward Fluorimetric-Paired-Emitter-Detector-Diode test for Bacillus anthracis DNA based on graphene oxide

Robert Ziółkowski , Sławomir Oszwałdowski , Konstancja Klaudia Kopyra , Katarzyna Zacharczuk , Aleksandra Anna Zasada , Elżbieta Malinowska

Abstract

The presented study is focused on the development of a cost-efficient and fully portable test for the detection of Bacillus anthracis pagA gene fragment as a real asymmetric PCR product. The complete detection procedure lasts only 2 min. with high selectivity and can be fully performed outside of the laboratory. The mass-produced parts (LEDs, PCR tube, multimeter and 2.5 V battery) make such a test also exceptionally cost-efficient. The key interaction used is that of graphene oxide's (GO) high affinity towards single-stranded DNA and its ability to quench the fluorescence. Initial optimization was conducted for HPLC purified cDNA. The test response was significantly dependent on GO concentration. The detected asymmetric PCR product was partly of double-stranded architecture which facilitates the test response. The developed Fluorimetric-Paired-Emitter-Detector-Diode (FPEDD) based DNA test allows for the detection of the pagA gene sequence at the level of 0.625 μM with 0.625–2.5 μM linear detection range. Both LOD and linear range can be improved in the case of longer assay times.

Author Robert Ziółkowski (FC / CMB)
Robert Ziółkowski,,
- Chair of Medical Biotechnology
, Sławomir Oszwałdowski (FC / CAC)
Sławomir Oszwałdowski,,
- Chair Of Analytical Chemistry
, Konstancja Klaudia Kopyra (FC / CMB)
Konstancja Klaudia Kopyra,,
- Chair of Medical Biotechnology
, Katarzyna Zacharczuk - [National Medicines Institute, Warsaw]
Katarzyna Zacharczuk,,
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, Aleksandra Anna Zasada - [National Medicines Institute, Warsaw]
Aleksandra Anna Zasada,,
-
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, Elżbieta Malinowska (FC / CMB)
Elżbieta Malinowska,,
- Chair of Medical Biotechnology
Journal seriesMicrochemical Journal, ISSN 0026-265X, e-ISSN 1095-9149
Issue year2020
Vol154
Pages104592_1-104592_9
Publication size in sheets0.3
ASJC Classification1602 Analytical Chemistry; 1607 Spectroscopy
DOIDOI:10.1016/j.microc.2019.104592
URL https://www.sciencedirect.com/science/article/pii/S0026265X19323598
Languageen angielski
File
wdpb_publikacje_pliki_plik_publikacja_3870_org.pdf 2.51 MB
Score (nominal)70
Score sourcejournalList
ScoreMinisterial score = 70.0, 07-08-2020, ArticleFromJournal
Publication indicators Scopus Citations = 0; Scopus SNIP (Source Normalised Impact per Paper): 2016 = 1.305; WoS Impact Factor: 2018 = 3.206 (2) - 2018=2.993 (5)
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