Rapid semi-automated quantitative multiplex tandem PCR (MT-PCR) assays for the differential diagnosis of influenza-like illness.
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.
Szewczuk
Elektra
E
Centre for Infectious Diseases and Microbiology, University of Sydney, Westmead Hospital, Westmead NSW 2145, Australia.
Thapa
Kiran
K
Anninos
Terry
T
McPhie
Kenneth
K
Higgins
Geoff
G
Dwyer
Dominic E
DE
Stanley
Keith K
KK
Iredell
Jonathan R
JR
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*
D006389:Hemagglutinins, Viral; C458881:hemagglutinin fusogenic peptide, influenza virus
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