Sandwich-Type DNA Micro-Optode Based on Gold–Latex Spheres Label for Reflectance Dengue Virus Detection

Sensors

Alizar Ulianas¹  Nurul Yuziana Mohd. Yusof²  Bahariah Khalid³  Nur Diyana Jamaluddin⁴  Jeningsih⁴  Goh Choo Ta⁴  Nur-Fadhilah Mazlan⁴  Ling Ling Tan⁴  Lee Yook Heng⁴ 

[1] Department of Chemistry, Faculty of Mathematics and Science, Universitas Negeri Padang, Padang 25131, Indonesia;Department of Earth Sciences and Environment, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi 43600, Malaysia;Department of Medicine, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400, Malaysia;Southeast Asia Disaster Prevention Research Initiative (SEADPRI-UKM), Institute for Environment and Development (LESTARI), Universiti Kebangsaan Malaysia, Bangi 43600, Malaysia;

关键词: gold nanoparticles;    latex particles;    optical biosensor;    reflectance;    sandwich hybridization;   

DOI  :  10.3390/s20071820

来源: DOAJ

【 摘 要 】

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−latex spheres were attached to the thiolated reporter probe (rDNA) by Au−thiol binding to functionalize as an optical gold−latex−rDNA label. The one-step sandwich hybridization recognition involved a pair of a DNA probe, i.e., capture probe (pDNA), and AuNP−PSA reporter label that flanked the target DNA (complementary DNA (cDNA)). The concentration of dengue virus cDNA was optically transduced by immobilized AuNP−PSA−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 × 10⁻²¹ M to 1.0 × 10⁻¹² M cDNA (R² = 0.9807) with a limit of detection (LOD) of 1 × 10⁻²⁹ 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.

【 授权许可】

Unknown