title: Reverse transcription strand invasion based amplification (RT-SIBA): a method for rapid detection of influenza A and B.
abstract: Rapid and accurate diagnosis of influenza viruses plays an important role in infection control, as well as in preventing the misuse of antibiotics. Isothermal nucleic acid amplification methods offer significant advantages over the polymerase chain reaction (PCR), since they are more rapid and do not require the sophisticated instruments needed for thermal cycling. We previously described a novel isothermal nucleic acid amplification method, 'Strand Invasion Based Amplification' (SIBA®), with high analytical sensitivity and specificity, for the detection of DNA. In this study, we describe the development of a variant of the SIBA method, namely, reverse transcription SIBA (RT-SIBA), for the rapid detection of viral RNA targets. The RT-SIBA method includes a reverse transcriptase enzyme that allows one-step reverse transcription of RNA to complementary DNA (cDNA) and simultaneous amplification and detection of the cDNA by SIBA under isothermal reaction conditions. The RT-SIBA method was found to be more sensitive than PCR for the detection of influenza A and B and could detect 100 copies of influenza RNA within 15 min. The development of RT-SIBA will enable rapid and accurate diagnosis of viral RNA targets within point-of-care or central laboratory settings.
authors.affiliation: Research and Development, Orion Diagnostica Oy, P. O. BOX 83, FI-02101, Espoo, Finland. kevin.eboigbodin@oriondiagnostica.fi.
authors.forename: Kevin
authors.initials: K
authors.lastname: Eboigbodin
authors.affiliation: Research and Development, Orion Diagnostica Oy, P. O. BOX 83, FI-02101, Espoo, Finland.
authors.forename: Sanna
authors.initials: S
authors.lastname: Filén
authors.affiliation: Research and Development, Orion Diagnostica Oy, P. O. BOX 83, FI-02101, Espoo, Finland.
authors.forename: Tuomas
authors.initials: T
authors.lastname: Ojalehto
authors.affiliation: Research and Development, Orion Diagnostica Oy, P. O. BOX 83, FI-02101, Espoo, Finland.
authors.forename: Mirko
authors.initials: M
authors.lastname: Brummer
authors.affiliation: Research and Development, Orion Diagnostica Oy, P. O. BOX 83, FI-02101, Espoo, Finland.
authors.forename: Sonja
authors.initials: S
authors.lastname: Elf
authors.affiliation: Research and Development, Orion Diagnostica Oy, P. O. BOX 83, FI-02101, Espoo, Finland.
authors.forename: Kirsi
authors.initials: K
authors.lastname: Pousi
authors.affiliation: Molecular Biology, GeneForm Technologies, Broadstairs, UK.
authors.forename: Mark
authors.initials: M
authors.lastname: Hoser
mesh_terms: D017931:DNA Primers; D006801:Humans; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification*; D009981:Influenza B virus / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis*; D025202:Molecular Diagnostic Techniques; D021141:Nucleic Acid Amplification Techniques; D019095:Point-of-Care Systems; D012367:RNA, Viral / Q000235:genetics; D048348:Reverse Transcription; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D012367:RNA, Viral
keywords: Amplification; Diagnostics; Influenza; Isothermal; RNA; Virus