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Annals of Clinical & Laboratory Science 39:155-159 (2009)
© 2009 Association of Clinical Scientists

Validation of the TaqMan Influenza A Detection Kit and a Rapid Automated Total Nucleic Acid Extraction Method to Detect Influenza A Virus in Nasopharyngeal Specimens

Shelly Bolotin1, Cedric De Lima1, Kam-Wing Choi1, Ernesto Lombos1, Laura Burton1, Tony Mazzulli1,2,3 and Steven J. Drews1,2,3
1 Ontario Public Health Laboratories, 2 Department of Microbiology, Mount Sinai Hospital, and 3 Department of Laboratory Medicine & Pathobiology, University of Toronto, Toronto, Ontario, Canada.

Address correspondence to Dr. Shelly Bolotin, Ontario Public Health Laboratories, Molecular Diagnostics, 81 Resources Road, Etobiocoke, Ontario M9P 3T1, Canada; tel 416 235 6504; fax 416 235 6550; e-mail shelly.bolotin{at}ontario.ca.

This study describes the validation of the TaqMan® Influenza A Detection Kit v2.0 combined with an automated nucleic acid extraction method. The limit of detection of this assay was determined by probit regression (95% confidence interval) to be 2 influenza A/PR/8/34 (H1N1) virus particles per microlitre. One hundred and eleven specimens previously tested using the Seeplex® RV assay and viral culture methods were tested using the TaqMan Influenza A Detection Kit. Compared to the aggregate gold-standard, the sensitivity and specificity of the TaqMan Influenza A Detection Kit were 100% (35/35) and 97% (74/76), respectively. Because of its accuracy, quick turn-around-time and lyophilized bead form, the TaqMan Influenza A Detection Kit, combined with the NucliSense® easyMAGTM automated extraction method, constitutes a reliable protocol for influenza A diagnosis.

Keywords: influenza, respiratory viruses, pandemic preparedness, molecular diagnostics




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