Abstract
A sensitive and selective optical DNA biosensor was developed for dengue virus detection based on novel square-planar piperidine side chain-functionalized N,N′-bis-4-(hydroxysalicylidene)-phenylenediamine-nickel(II), which was able to intercalate via nucleobase stacking within DNA and be functionalized as an optical DNA hybridization marker. 3-Aminopropyltriethoxysilane (APTS)-modified porous silica nanospheres (PSiNs), was synthesized with a facile mini-emulsion method to act as a high capacity DNA carrier matrix. The Schiff base salphen complexes-labelled probe to target nucleic acid on the PSiNs renders a colour change of the DNA biosensor to a yellow background colour, which could be quantified via a reflectance transduction method. The reflectometric DNA biosensor demonstrated a wide linear response range to target DNA over the concentration range of 1.0 × 10−16–1.0 × 10−10 M (R2 = 0.9879) with an ultralow limit of detection (LOD) at 0.2 aM. The optical DNA biosensor response was stable and maintainable at 92.8% of its initial response for up to seven days of storage duration with a response time of 90 min. The reflectance DNA biosensor obtained promising recovery values of close to 100% for the detection of spiked synthetic dengue virus serotypes 2 (DENV-2) DNA concentration in non-invasive human samples, indicating the high accuracy of the proposed DNA analytical method for early diagnosis of all potential infectious diseases or pathological genotypes.
Highlights
Dengue fever has become a major public health concern in Malaysia and many other tropical countries [1]
It is presumed that the as-synthesized piperidine side chain functionalized nickel(II) salphen complex was bound to the double-stranded DNA (dsDNA) by intercalation between base pairs ascribed to the square-planar coordination geometry structured of the Schiff base ligand bonded to the metal centre that enhances its ability to intercalate between dsDNA [16]
The described reflectance dengue virus DNA biosensor was applied to determine the known synthetic complementary DNA (cDNA) concentration spiked into non-invasive human bodily fluid samples such as urine and saliva, as dengue virus infection in human has been reported to be detected in both urine and saliva but not in plasma [33]
Summary
Dengue fever has become a major public health concern in Malaysia and many other tropical countries [1]. In order to improve the selectivity of the biosensor towards the dengue virus, DNA is an ideal target for specific detection of viral pathogens. Zhang and co-workers [4] have constructed an innovative field-effect transistor (FET)-based DNA biosensor for highly sensitive and rapid detection of the dengue virus. The fabrication of the silicon nanowire transducer required etching and oxidation of a silicon-on-insulator wafer at high temperature, and the tedious need for further lithographic methods Another electrochemical DNA-based biosensor for the detection of dengue serotypes at picomolar concentration level was fabricated using gold nanoparticles-polyaniline hybrid composites. As the proposed yellow coloured Schiff base metal complex gave a colour change during the probe to target DNA hybridization reaction, an optical dengue virus DNA biosensor was developed based on light reflection transduction to quantify the corresponding targeted DNA concentration. The resulting solid-state DNA biosensor allowed visual detection of dengue viruses by the human naked eye by visual observing the colour change at the surface of the PSiNs-based DNA biosensing platform
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