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.
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