title: VereFlu™: an integrated multiplex RT-PCR and microarray assay for rapid detection and identification of human influenza A and B viruses using lab-on-chip technology.
abstract: Threatening sporadic outbreaks of avian influenza and the H1N1 pandemic of 2009 highlight the need for rapid and accurate detection and typing of influenza viruses. In this paper, we describe the validation of the VereFlu™ Lab-on-Chip Influenza Assay, which is based on the integration of two technologies: multiplex reverse transcription (RT)-PCR followed by microarray amplicon detection. This assay simultaneously detects five influenza virus subtypes, including the 2009 pandemic influenza A (H1N1), seasonal H1N1, H3N2, H5N1 and influenza B virus. The VereFlu™ assay was clinically validated in Singapore and compared against reference methods of real-time PCR, virus detection by immunofluorescence of cell cultures and sequencing. A sensitivity and specificity of 96.8% and 92.8%, respectively, was demonstrated for pandemic H1N1; 95.7% and 100%, respectively, for seasonal H1N1; 91.2% and 97.6%, respectively, for seasonal H3N2; 95.2% and 100%, respectively, for influenza B. Additional evaluations carried out at the World Health Organization (WHO) Collaborating Centre, Melbourne, Australia, confirmed that the test was able to reliably detect H5N1. This portable, fast time-to-answer (3 hours) device is particularly suited for diagnostic applications of detection, differentiation and identification of human influenza virus subtypes.
authors.affiliation: Department of Laboratory Medicine, National University Hospital, Singapore, 119074, Singapore. Jeanette_Teo@nuhs.edu.sg
authors.forename: Jeanette
authors.initials: J
authors.lastname: Teo
authors.forename: Patrizia
authors.initials: P
authors.lastname: Di Pietro
authors.forename: Floriana
authors.initials: F
authors.lastname: San Biagio
authors.forename: Monica
authors.initials: M
authors.lastname: Capozzoli
authors.forename: Yi-Mo
authors.initials: YM
authors.lastname: Deng
authors.forename: Ian
authors.initials: I
authors.lastname: Barr
authors.forename: Natalie
authors.initials: N
authors.lastname: Caldwell
authors.forename: Kian-Leong
authors.initials: KL
authors.lastname: Ong
authors.forename: Mitsuharu
authors.initials: M
authors.lastname: Sato
authors.forename: Rosemary
authors.initials: R
authors.lastname: Tan
authors.forename: Raymond
authors.initials: R
authors.lastname: Lin
mesh_terms: D006801:Humans; D009980:Influenza A virus / Q000145:classification / Q000302:isolation & purification*; D009981:Influenza B virus / Q000302:isolation & purification*; D056656:Lab-On-A-Chip Devices; D040081:Protein Array Analysis / Q000379:methods*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity