Abstract
A rapid and confident tool to identify and diagnose bacterial pathogens with more accuracy using DNA as fingerprints is necessary. Herein, we report a smart chemosensor having a terminal adenine sticking to the thymine of single-stranded DNA (ssDNA) through supramolecular interactions and, which leaves ssDNA when the same ssDNA matches with the targeting desired DNA. We have synthesized a naked-eye coloured chemosensor with carbazole. As a model genetic material, DNA of Clavibacter michiganensis subsp. michiganensis was hybridized to ssDNA and immobilized over nitrocellulose membrane. The prepared adenine-chemosensor, by passing through the nitrocellulose-ssDNA membrane caused the formation of ssDNA nitrocellulose-ssDNA-adenine-chemosensor. FTIR results of the immobilized ssDNAs showed that the matching of same ssDNA releases the adenine-chemosensor from the surface of nitrocellulose-ssDNA that results in formation of the double stranded DNA. The selectivity of chemosensor was also confirmed with different bacterial DNA (Bacillus subtilis) as control. These data highlights accurate and reliable results of a new diagnostic kit prototype promising for further studies, which is able to diagnose DNA quickly and precisely.
Highlights
Biosensors have recently become efficient analytical tools attracting the attention of researchers due to their easy, convenient and specific diagnosis potential compared to traditional methods
The aim of our study is to synthesize molecules that can act as sensors binding to single-stranded DNA, and to make fast and sensitive DNA diagnosis with the help of Fourier-transform infrared spectroscopy (FTIR) spectra to be obtained based on the hybridization of these molecules with target DNA
The peaks of the methylene (-CH2-) groups in the structure of the synthesized BSP-3 chemosensor were determined by hydrogen-1 NMR (HNMR) analysis (Figure 2a)
Summary
Biosensors have recently become efficient analytical tools attracting the attention of researchers due to their easy, convenient and specific diagnosis potential compared to traditional methods. DNA-based method “DNA sensors or gene sensors” are used for diagnosis and the detection of the targeted region.[1,2,3] These biosensors usually relies on detecting the hybridization of the target ssDNA strand, which are complementary strands.[1,2,4] The advances in molecular biology have become the reason for increasing the interest in the design of DNA hybridization-based biosensors.[5,6,7,8,9] DNA-based biosensors can be used in many areas such as clinical, environmental and food safety issues. Nucleotide sequences called primers or probes are compatible with bacterial DNA and hybrid molecules formed after adaptation. The hybrid detected with the presence of a reporter molecule provides signal amplification, while the product resulting from enzymatic amplification of the hybrid is detected indirectly.[10]
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