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

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.

[1]  Neus G Bastús,et al.  Kinetically controlled seeded growth synthesis of citrate-stabilized gold nanoparticles of up to 200 nm: size focusing versus Ostwald ripening. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[2]  Varun Rai,et al.  Ultrasensitive cDNA Detection of Dengue Virus RNA Using Electrochemical Nanoporous Membrane-Based Biosensor , 2012, PloS one.

[3]  P. Chomczyński,et al.  Effect of pH and ionic strength on the spectrophotometric assessment of nucleic acid purity. , 1997, BioTechniques.

[4]  Li Shen,et al.  Point-of-care colorimetric detection with a smartphone. , 2012, Lab on a chip.

[5]  W. Siegert,et al.  Guideline to reference gene selection for quantitative real-time PCR. , 2004, Biochemical and biophysical research communications.

[6]  Richard A Montagna,et al.  Microfluidic biosensor for the serotype-specific detection of dengue virus RNA. , 2005, Analytical chemistry.

[7]  M. Deutscher,et al.  RNA quality control: degradation of defective transfer RNA , 2002, The EMBO journal.

[8]  Genxi Li,et al.  A Centrifugation-based Method for Preparation of Gold Nanoparticles and its Application in Biodetection , 2007, International Journal of Molecular Sciences.

[9]  R. Meagher,et al.  Colorimetric-Luminance Readout for Quantitative Analysis of Fluorescence Signals with a Smartphone CMOS Sensor. , 2018, Analytical chemistry.

[10]  Li Zhang,et al.  Silicon nanowire biosensor for highly sensitive and rapid detection of Dengue virus , 2010 .

[11]  Sara W. Bird,et al.  A smartphone-based diagnostic platform for rapid detection of Zika, chikungunya, and dengue viruses , 2017, Scientific Reports.

[12]  M. Ravichandran,et al.  Gold-nanoparticle based electrochemical DNA sensor for the detection of fish pathogen Aphanomyces invadans. , 2013, Talanta.

[13]  Sook Mei Khor,et al.  An introduction to dengue-disease diagnostics , 2015 .

[14]  D. Fuchs,et al.  Detection of serum neopterin for early assessment of dengue virus infection , 2006, Journal of Infection.

[15]  S. Tajima,et al.  Confirmation of dengue virus infection by detection of dengue virus type 1 genome in urine and saliva but not in plasma. , 2007, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[16]  Ahmad Musa,et al.  A Biosensor for Urea from Succinimide-Modified Acrylic Microspheres Based on Reflectance Transduction , 2011, Sensors.

[17]  Yook Heng Lee,et al.  An Electrochemical DNA Microbiosensor Based on Succinimide-Modified Acrylic Microspheres , 2012, Sensors.

[18]  Om Parkash,et al.  Diagnosis of Dengue Infection Using Conventional and Biosensor Based Techniques , 2015, Viruses.

[19]  M. H. Simonian,et al.  Spectrophotometric and Colorimetric Determination of Protein Concentration , 1996, Current protocols in molecular biology.

[20]  A. S. Milligan,et al.  Toward specific detection of Dengue virus serotypes using a novel modular biosensor. , 2010, Biosensors & bioelectronics.

[21]  A. Baeumner,et al.  Biosensor for dengue virus detection: sensitive, rapid, and serotype specific. , 2002, Analytical chemistry.

[22]  V. Stollar,et al.  Studies on the nature of dengue viruses. III. RNA synthesis in cells infected with type 2 dengue virus. , 1967, Virology.

[23]  Chee Seng Toh,et al.  Impedimetric DNA Biosensor Based on a Nanoporous Alumina Membrane for the Detection of the Specific Oligonucleotide Sequence of Dengue Virus , 2013, Sensors.

[24]  C. P. de Melo,et al.  An impedimetric biosensor for detection of dengue serotype at picomolar concentration based on gold nanoparticles-polyaniline hybrid composites. , 2011, Colloids and surfaces. B, Biointerfaces.

[25]  Jia Shin Ho,et al.  Development of an electrochemical membrane-based nanobiosensor for ultrasensitive detection of dengue virus. , 2012, Analytica chimica acta.

[26]  Dong-Eun Kim,et al.  Desorption of single-stranded nucleic acids from graphene oxide by disruption of hydrogen bonding. , 2013, The Analyst.