title: Development and preliminary evaluation of a rapid oligochromatographic assay for specific detection of new human influenza A H1N1 virus.
abstract: A new oligochromatographic assay, Speed-Oligo Novel Influenza A H1N1, was designed and optimized for the specific detection of the 2009 influenza A H1N1 virus. The assay is based on a PCR method coupled to detection of PCR products by means of a dipstick device. The target sequence is a 103-bp fragment within the hemagglutinin gene. The analytical sensitivity of the new assay was measured with serial dilutions of a plasmid that contained the target sequence, and we determined that down to one copy per reaction of the plasmid was reliably detected. Diagnostic performance was assessed with 103 RNAs from suspected cases (40 positive and 63 negative results) previously analyzed with a reference real-time PCR technique. All positive cases were confirmed, and no false-positive results were detected with the new assay. No cross-reactions were observed when other viral strains or clinical samples with other respiratory viruses were tested. According to these results, this new assay has 100% sensitivity and specificity. The turnaround time for the whole procedure was 140 min. The assay may be especially useful for the specific detection of 2009 H1N1 virus in laboratories not equipped with real-time PCR instruments.
authors.affiliation: Servicio de Microbiología, Hospital Universitario Virgen de las Nieves, Avda. Fuerzas Armadas 2, 18014 Granada, Spain. mercedes.perez.ruiz.sspa@juntadeandalucia.es
authors.forename: Mercedes
authors.initials: M
authors.lastname: Pérez-Ruiz
authors.forename: José-María
authors.initials: JM
authors.lastname: Navarro-Marí
authors.forename: María-Fé
authors.initials: MF
authors.lastname: Bautista-Marín
authors.forename: Irene
authors.initials: I
authors.lastname: Pedrosa-Corral
authors.forename: Sara
authors.initials: S
authors.lastname: Sanbonmatsu-Gámez
authors.forename: Ana G
authors.initials: AG
authors.lastname: Camacho
authors.forename: José
authors.initials: J
authors.lastname: Rojas
authors.forename: Jorge
authors.initials: J
authors.lastname: Ruiz-Ortiz
authors.forename: Javier
authors.initials: J
authors.lastname: Rodríguez-Granger
authors.forename: José Antonio
authors.initials: JA
authors.lastname: Carrillo
mesh_terms: D002845:Chromatography / Q000379:methods*; D003952:Diagnostic Imaging / Q000706:statistics & numerical data; D006389:Hemagglutinins, Viral / Q000235:genetics; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D010957:Plasmids; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014773:Virology / Q000379:methods*
chemical_list: D006389:Hemagglutinins, Viral; D012367:RNA, Viral; C458881:hemagglutinin fusogenic peptide, influenza virus