1. Sandwich-Type DNA Micro-Optode Based on Gold–Latex Spheres Label for Reflectance Dengue Virus Detection
- Author
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Nur-Fadhilah Mazlan, Nurul Yuziana Mohd Yusof, Goh Choo Ta, Nur Diyana Jamaluddin, Alizar Ulianas, Jeningsih, Ling Ling Tan, Bahariah Khalid, and Lee Yook Heng
- Subjects
reflectance ,Polymers ,Metal Nanoparticles ,Succinimides ,02 engineering and technology ,Biosensing Techniques ,Dengue virus ,lcsh:Chemical technology ,medicine.disease_cause ,01 natural sciences ,Biochemistry ,Article ,Analytical Chemistry ,Dengue ,chemistry.chemical_compound ,Complementary DNA ,medicine ,sandwich hybridization ,Humans ,lcsh:TP1-1185 ,A-DNA ,Electrical and Electronic Engineering ,Instrumentation ,Detection limit ,Chromatography ,optical biosensor ,latex particles ,010401 analytical chemistry ,Dengue Virus ,021001 nanoscience & nanotechnology ,Atomic and Molecular Physics, and Optics ,Microspheres ,0104 chemical sciences ,Reverse transcription polymerase chain reaction ,chemistry ,Acrylates ,Colloidal gold ,gold nanoparticles ,DNA, Viral ,Gold ,0210 nano-technology ,Biosensor ,DNA - Abstract
A DNA micro-optode for dengue virus detection was developed based on the sandwich hybridization strategy of DNAs on succinimide-functionalized poly(n-butyl acrylate) (poly(nBA-NAS)) microspheres. Gold nanoparticles (AuNPs) with an average diameter of ~20 nm were synthesized using a centrifugation-based method and adsorbed on the submicrometer-sized polyelectrolyte-coated poly(styrene-co-acrylic acid) (PSA) latex particles via an electrostatic method. The AuNP&ndash, latex spheres were attached to the thiolated reporter probe (rDNA) by Au&ndash, thiol binding to functionalize as an optical gold&ndash, latex&ndash, rDNA label. The one-step sandwich hybridization recognition involved a pair of a DNA probe, i.e., capture probe (pDNA), and AuNP&ndash, PSA reporter label that flanked the target DNA (complementary DNA (cDNA)). The concentration of dengue virus cDNA was optically transduced by immobilized AuNP&ndash, PSA&ndash, rDNA conjugates as the DNA micro-optode exhibited a violet hue upon the DNA sandwich hybridization reaction, which could be monitored by a fiber-optic reflectance spectrophotometer at 637 nm. The optical genosensor showed a linear reflectance response over a wide cDNA concentration range from 1.0 ×, 10&minus, 21 M to 1.0 ×, 12 M cDNA (R2 = 0.9807) with a limit of detection (LOD) of 1 ×, 29 M. The DNA biosensor was reusable for three consecutive applications after regeneration with mild sodium hydroxide. The sandwich-type optical biosensor was well validated with a molecular reverse transcription polymerase chain reaction (RT-PCR) technique for screening of dengue virus in clinical samples, e.g., serum, urine, and saliva from dengue virus-infected patients under informed consent.
- Published
- 2020
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