title: Direct quantitative screening of influenza A virus without DNA amplification by single-particle dual-mode total internal reflection scattering.
abstract: Quantitative screening of influenza A (H7N9) virus without DNA amplification was performed based on single-particle dual-mode total internal reflection scattering (SD-TIRS) with a transmission grating (TG). A gold nanopad was utilized as a substrate for the hybridization of probe DNA molecules with the TIRS nanotag (silver-nanoparticle). The TG effectively isolated the scattering signals in first-order spectral images (n=+1) of the nanotag from that of the substrate, providing excellent enhancement of signal-to-noise and selectivity. By using single-DNA molecule/TIRS nanotag hybridization, target DNA molecules of H7N9 were detected down to 74 zM, which is at least 100,000 times lower than the current detection limit of 9.4fM. By simply modifying the design of the probe DNA molecules, this technique can be used to directly screen other viral DNAs in various human biological samples at the single-molecule level without target amplification.
authors.affiliation: Department of Applied Chemistry and Institute of Natural Sciences, College of Applied Science, Kyung Hee University, Yongin-si, Gyeonggi-do 17104, Republic of Korea.
authors.forename: Seungah
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Graduate School, Kyung Hee University, Yongin-si, Gyeonggi-do 17104, Republic of Korea.
authors.forename: Suresh Kumar
authors.initials: SK
authors.lastname: Chakkarapani
authors.affiliation: Department of Chemistry, Iowa State University, Ames, IA 50011, USA.
authors.forename: Edward S
authors.initials: ES
authors.lastname: Yeung
authors.affiliation: Department of Applied Chemistry and Institute of Natural Sciences, College of Applied Science, Kyung Hee University, Yongin-si, Gyeonggi-do 17104, Republic of Korea; Department of Chemistry, Graduate School, Kyung Hee University, Yongin-si, Gyeonggi-do 17104, Republic of Korea. Electronic address: shkang@khu.ac.kr.
authors.forename: Seong Ho
authors.initials: SH
authors.lastname: Kang
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D004867:Equipment Design; D006046:Gold / Q000737:chemistry; D006801:Humans; D064766:Influenza A Virus, H7N9 Subtype / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000821:virology*; D053768:Metal Nanoparticles / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D049329:Nanostructures / Q000737:chemistry; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012834:Silver / Q000737:chemistry
chemical_list: D015342:DNA Probes; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D012834:Silver; D006046:Gold
keywords: Amplification-free; Influenza A virus; Single-molecule spectroscopy; Total internal reflection scattering; Viral DNA