tetano
Editor, Senior Moderator
J Clin Microbiol. 2010 Jul 7. [Epub ahead of print]
Analytical Performance Determination and Clinical Validation of the Novel Roche RealTime ready Influenza A/H1N1 Detection Set.
Wenzel JJ, Panning M, Kaul KL, Mangold KA, Revell PA, Luna RA, Zepeda H, Perea L, Vazquez-Perez JA, Young S, Rodic-Polic B, Eickmann M, Drosten C, Jilg W, Reischl U.
Institute of Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany; Department of Virology, University of Freiburg, Freiburg, Germany; NorthShore University Health System Research Institute, Evanston, Illinois, U.S.A.; Department of Pathology, Texas Children's Hospital, Baylor College of Medicine, Houston, Texas, U.S.A.; Laboratorio Medicina de Conservaci?n, Escuela Superior de Medicina, Instituto Polit?cnico Nacional, Colonia de Santo Tom?s, M?xico; Centro de Investigaci?n en Enfermedades Infecciosas, Instituto Nacional de Enfermedades Respiratorias, Tlalpan, M?xico; Department of Pathology, University of New Mexico School of Medicine, Albuquerque, New Mexico, U.S.A.; Institute for Virology, University of Marburg, Marburg, Germany; Institute of Virology, University of Bonn Medical Centre, Bonn, Germany.
Abstract
The emergence of a novel pandemic human strain of influenza A (H1N1/09) in April 2009 has demonstrated the need for well-validated diagnostic tests which are broadly applicable, rapid, sensitive and specific. The analytical performance and clinical validity of results generated with the novel Roche RealTime ready Influenza A/H1N1 Detection Set using the LightCycler(R) 2.0 Instrument was characterized. Analytical performance was assessed by processing respiratory samples spiked with H1N1/09 and seasonal influenza A virus, a set of seasonal influenza A virus subtypes and samples containing common viral and bacterial respiratory pathogens. Clinical validity of results was assessed in comparison to other assays by analyzing 359 specimens at three clinical sites and one reference laboratory. Direct sequencing was used to resolve samples with discrepant results. The assay detected virus concentrations down to <50 RNA copies per RT-qPCR reaction. Various influenza A virus subtypes were covered. Analytical specificity was 100 %. High clinical validity was demonstrated by 99% positive agreement between seasonal influenza A, 98% between H1N1/09, and 88% agreement between negative results. Analytical sensitivity was compared to three other RT-qPCR assays and found to be equivalent. The novel Roche RealTime ready Influenza A/H1N1 Detection Set can be utilized on the widely-used LightCycler(R) platform. We demonstrate its usefulness for rapid detection and surveillance of pandemic H1N1/09 influenza A virus infections.
PMID: 20610677 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/20610677
Analytical Performance Determination and Clinical Validation of the Novel Roche RealTime ready Influenza A/H1N1 Detection Set.
Wenzel JJ, Panning M, Kaul KL, Mangold KA, Revell PA, Luna RA, Zepeda H, Perea L, Vazquez-Perez JA, Young S, Rodic-Polic B, Eickmann M, Drosten C, Jilg W, Reischl U.
Institute of Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany; Department of Virology, University of Freiburg, Freiburg, Germany; NorthShore University Health System Research Institute, Evanston, Illinois, U.S.A.; Department of Pathology, Texas Children's Hospital, Baylor College of Medicine, Houston, Texas, U.S.A.; Laboratorio Medicina de Conservaci?n, Escuela Superior de Medicina, Instituto Polit?cnico Nacional, Colonia de Santo Tom?s, M?xico; Centro de Investigaci?n en Enfermedades Infecciosas, Instituto Nacional de Enfermedades Respiratorias, Tlalpan, M?xico; Department of Pathology, University of New Mexico School of Medicine, Albuquerque, New Mexico, U.S.A.; Institute for Virology, University of Marburg, Marburg, Germany; Institute of Virology, University of Bonn Medical Centre, Bonn, Germany.
Abstract
The emergence of a novel pandemic human strain of influenza A (H1N1/09) in April 2009 has demonstrated the need for well-validated diagnostic tests which are broadly applicable, rapid, sensitive and specific. The analytical performance and clinical validity of results generated with the novel Roche RealTime ready Influenza A/H1N1 Detection Set using the LightCycler(R) 2.0 Instrument was characterized. Analytical performance was assessed by processing respiratory samples spiked with H1N1/09 and seasonal influenza A virus, a set of seasonal influenza A virus subtypes and samples containing common viral and bacterial respiratory pathogens. Clinical validity of results was assessed in comparison to other assays by analyzing 359 specimens at three clinical sites and one reference laboratory. Direct sequencing was used to resolve samples with discrepant results. The assay detected virus concentrations down to <50 RNA copies per RT-qPCR reaction. Various influenza A virus subtypes were covered. Analytical specificity was 100 %. High clinical validity was demonstrated by 99% positive agreement between seasonal influenza A, 98% between H1N1/09, and 88% agreement between negative results. Analytical sensitivity was compared to three other RT-qPCR assays and found to be equivalent. The novel Roche RealTime ready Influenza A/H1N1 Detection Set can be utilized on the widely-used LightCycler(R) platform. We demonstrate its usefulness for rapid detection and surveillance of pandemic H1N1/09 influenza A virus infections.
PMID: 20610677 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/20610677