title: Simultaneous detection of influenza viruses A, B, and swine origin influenza A using multiplex one-step real-time RT-PCR assay.
abstract: Every year, seasonal epidemics of influenza viruses are causing considerable morbidity and mortality worldwide. Also infrequent novel and rearranged strains of influenza viruses have caused quick, acute universal pandemics resulting in millions of mortalities. The usage of efficient and accurate detection is superior for infection control, effective treatment, and epidemiological supervision. Therefore, evaluation of useful real-time PCR molecular tests for the detection of pandemic viruses is important before the next wave of the pandemic. A novel quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) assay with specific primers was used successfully for detection and monitoring of the influenza A, B, and swine influenza. The newly designed primers target highly conserved regions in influenza viruses. Our qRT-PCR assay is highly specific for detecting influenza A, B, and swine influenza viruses. The cutoff CT value was determined <38 for domestic human diagnostic test, under conditions of FDA emergency, and the reaction efficiency of the InfA, swInfA, and InfB assays were thereby estimated to be 97.9 % (R2 = 0.998), 98.3 % (R2 = 0.986), and 99.5 % (R2 = 0.995), respectively. Interestingly, based on our finding, there is no cross reactivity of detecting other viruses.
authors.affiliation: Department of Virology and Anti Microbial Resistance Research Center, Iran University of Medical Sciences, Hemmat Highway, Tehran, Iran, hrmonavari@yahoo.com.
authors.forename: S H R
authors.initials: SH
authors.lastname: Monavari
authors.forename: H R
authors.initials: HR
authors.lastname: Mollaie
authors.forename: M
authors.initials: M
authors.lastname: Fazlalipour
mesh_terms: D000818:Animals; D004285:Dogs; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D009981:Influenza B virus / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000473:pathology / Q000821:virology; D057230:Limit of Detection; D061985:Madin Darby Canine Kidney Cells; D009976:Orthomyxoviridae Infections / Q000473:pathology / Q000821:virology; D011933:Reagent Kits, Diagnostic; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine / Q000821:virology*
chemical_list: D011933:Reagent Kits, Diagnostic