title: Rapid semi-automated quantitative multiplex tandem PCR (MT-PCR) assays for the differential diagnosis of influenza-like illness.
abstract: BACKGROUND
  Influenza A, including avian influenza, is a major public health threat in developed and developing countries. Rapid and accurate detection is a key component of strategies to contain spread of infection, and the efficient diagnosis of influenza-like-illness is essential to protect health infrastructure in the event of a major influenza outbreak.
  
  
  METHODS
  We developed a multiplexed PCR (MT-PCR) assay for the simultaneous diagnosis of respiratory viruses causing influenza-like illness, including the specific recognition of influenza A haemagglutinin subtypes H1, H3, and H5. We tested several hundred clinical specimens in two diagnostic reference laboratories and compared the results with standard techniques.
  
  
  RESULTS
  The sensitivity and specificity of these assays was higher than individual assays based on direct antigen detection and standard PCR against a range of control templates and in several hundred clinical specimens. The MT-PCR assays provided differential diagnoses as well as potentially useful quantitation of virus in clinical samples.
  
  
  CONCLUSIONS
  MT-PCR is a potentially powerful tool for the differential diagnosis of influenza-like illness in the clinical diagnostic laboratory.
authors.affiliation: Centre for Infectious Diseases and Microbiology, University of Sydney, Westmead Hospital, Westmead NSW 2145, Australia.
authors.forename: Elektra
authors.initials: E
authors.lastname: Szewczuk
authors.forename: Kiran
authors.initials: K
authors.lastname: Thapa
authors.forename: Terry
authors.initials: T
authors.lastname: Anninos
authors.forename: Kenneth
authors.initials: K
authors.lastname: McPhie
authors.forename: Geoff
authors.initials: G
authors.lastname: Higgins
authors.forename: Dominic E
authors.initials: DE
authors.lastname: Dwyer
authors.forename: Keith K
authors.initials: KK
authors.lastname: Stanley
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Iredell
mesh_terms: D003937:Diagnosis, Differential; D006389:Hemagglutinins, Viral / Q000235:genetics; D006801:Humans; D009980:Influenza A virus / Q000235:genetics; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
chemical_list: D006389:Hemagglutinins, Viral; C458881:hemagglutinin fusogenic peptide, influenza virus
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