title: Simultaneous discrimination and detection of influenza A(H1N1)pdm09 and seasonal influenza A viruses using a rapid immunogold biosensor.
abstract: A rapid immunogold biosensor for the simultaneous discrimination of influenza A(H1N1)pdm09 and seasonal influenza A viruses was developed successfully. Monoclonal antibodies (mAbs) that were specific for the hemagglutinin protein of the A(H1N1)pdm09 virus were produced, and the best mAb pairs were selected. Using an mAb that was specific for the influenza A nucleoprotein, a rapid immunogold biosensor for the discrimination and detection of A(H1N1)pdm09/seasonal influenza viruses was developed. When tested with 72 virus isolates, the system achieved 100 % detection of the A(H1N1)pdm09 virus without cross-reactivity against seasonal influenza A (H1, H3 subtypes) and B viruses, parainfluenza viruses, respiratory syncytial viruses, and adenoviruses. The detection limits for A(H1N1)pdm09 and seasonal strains were 5 × 10(2)-7.5 × 10(3) and 1 × 10(3)-7.5 × 10(5) TCID50/mL, respectively. When tested with 49 clinical specimens, the specificity was high (100 %). The sensitivity for the detection of A(H1N1)pdm09 and seasonal strains was 90 % and 100 %, respectively, which correlated with the results of real-time reverse transcription polymerase chain reaction as a reference method. The ability of the system to detect and discriminate the A(H1N1)pdm09 strain from the seasonal strains suggests that this method may be beneficial for investigation of outbreaks and diagnostic applications. Furthermore, this method might be a useful platform for developing a rapid diagnostic system for the simultaneous discrimination of other influenza virus subtypes during future outbreaks.
authors.affiliation: National Nanotechnology Center (NANOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani, 12120, Thailand.
authors.forename: Chayachon
authors.initials: C
authors.lastname: Apiwat
authors.forename: Natpapas
authors.initials: N
authors.lastname: Wiriyachaiporn
authors.forename: Weerakanya
authors.initials: W
authors.lastname: Maneeprakorn
authors.forename: Tararaj
authors.initials: T
authors.lastname: Dharakul
authors.forename: Charin
authors.initials: C
authors.lastname: Thepthai
authors.forename: Pilaipan
authors.initials: P
authors.lastname: Puthavathana
authors.forename: Sontana
authors.initials: S
authors.lastname: Siritantikorn
authors.forename: Navin
authors.initials: N
authors.lastname: Horthongkham
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D015374:Biosensing Techniques / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D053118:Influenza A Virus, H1N1 Subtype / Q000145:classification / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D012621:Seasons; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral