title: SERS molecular sentinel for the RNA genetic marker of PB1-F2 protein in highly pathogenic avian influenza (HPAI) virus.
abstract: We have developed a simple and sensitive assay for the detection of the RNA genetic marker associated with high pathogenicity influenza (HPAI) virus. The assay constituted of an array of Raman label tagged hairpin-DNA immobilized on a surface-enhanced Raman scattering (SERS) active substrate as the molecular sentinel (MS) reporter. Upon incubation of the assay with the target RNA, the structure of the hairpin-DNA probe changed from stem-loop configuration (closed state) to DNA/RNA hybridization configuration (open state) so that the Raman label tag will be physically separated from the SERS substrate and induce a decrease of Raman scattering intensity. A metal film over nanosphere (MFON) substrate was developed with a SERS enhancement of about 1.7 × 10(5). Based on this MS-modified substrate, the SERS signal showed a linear relationship to the target RNA in the range of 0-60 attomoles and the limit of detect is 2.67 attomoles. The non-complementary RNA sequences control was also detected and no spectral response was observed. The sensing process only required a single hybridization step and post-hybridization washing could also be omitted. Given that this ultrasensitive biosensor assay is free of polymerase chain reaction (PCR) amplification, it would be a potential diagnostic tool for point-of-care HPAI virus detection.
authors.affiliation: Key Laboratory of New Molecular Diagnosis Techniques for Infectious Diseases, Institute of Radiation Medicine, 27 Taiping Road, Beijing, China.
authors.forename: Yuanfeng
authors.initials: Y
authors.lastname: Pang
authors.affiliation: Key Laboratory of New Molecular Diagnosis Techniques for Infectious Diseases, Institute of Radiation Medicine, 27 Taiping Road, Beijing, China.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory of New Molecular Diagnosis Techniques for Infectious Diseases, Institute of Radiation Medicine, 27 Taiping Road, Beijing, China. Electronic address: ruixiao203@sina.com.
authors.forename: Rui
authors.initials: R
authors.lastname: Xiao
authors.affiliation: Key Laboratory of New Molecular Diagnosis Techniques for Infectious Diseases, Institute of Radiation Medicine, 27 Taiping Road, Beijing, China. Electronic address: sqwang@bmi.ac.cn.
authors.forename: Shengqi
authors.initials: S
authors.lastname: Wang
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D001717:Birds / Q000821:virology; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D004867:Equipment Design; D005819:Genetic Markers; D009980:Influenza A virus / Q000235:genetics* / Q000302:isolation & purification; D005585:Influenza in Birds / Q000175:diagnosis / Q000821:virology*; D009570:Nitriles / Q000737:chemistry; D011722:Pyrethrins / Q000737:chemistry; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics*; D013059:Spectrum Analysis, Raman / Q000295:instrumentation*; D014764:Viral Proteins / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D015342:DNA Probes; D005819:Genetic Markers; D009570:Nitriles; C442816:PB1-F2 protein, Influenza A virus; D011722:Pyrethrins; D012367:RNA, Viral; D014764:Viral Proteins; C037304:cyhalothrin
keywords: Molecular sentinel; Point-of-care; RNA genetic marker; Surface-enhanced Raman scattering