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J Clin Microbiol. 2010 Sep 8. [Epub ahead of print]
Real-Time Detection of Influenza A, Influenza B and Respiratory Syncytial Virus A/B in Respiratory Specimens using Nanoparticle Probes.
Jannetto PJ, Buchan BW, Vaughan KA, Ledford JS, Anderson DK, Henley DC, Quigley NB, Ledeboer NA.
Department of Pathology, Medical College of Wisconsin, and Dynacare Laboratories, 9200 West Wisconsin Ave., Milwaukee, WI 53226, USA; Molecular Pathology Laboratory Network, Inc., 250 East Broadway, Maryville, TN 37804.
Abstract
Seasonal epidemics of influenza and respiratory syncytial virus are responsible for significant morbidity and mortality worldwide. Infrequently, novel or re-emergent strains of influenza A have caused rapid, severe global pandemics resulting in millions of fatalities. The ability to efficiently and accurately detect and differentiate respiratory viruses is paramount to effective treatment, infection control, and epidemiological surveillance. We evaluated the ability of two FDA cleared nucleic acid-based tests, the semi-automated Respiratory Virus Nucleic Acid Test (VRNAT) and the fully automated Respiratory Virus Nucleic Acid Test SP (RVNATSP) (Nanosphere Inc., Northbrook, IL) to detect influenza A, influenza B, and RSV A/B from clinical nasopharyngeal swab specimens. Detection of viral RNA in both tests is based on nucleic acid amplification followed by hybridization to capture probes immobilized on a glass slide. A novel technology utilizing gold nanoparticle conjugated probes is utilized to detect the presence of captured target DNA. This microarray-based approach to detection has proven to be more sensitive than traditional culture/DFA method for detecting RSV and influenza viruses in clinical specimens, including the novel 2009 H1N1 strain. Specifically, we report 98.0% sensitivity and 96.5% specificity of the VRNAT compared to culture/DFA. Further, the VRNAT detected virus in an additional 58% of specimens that were culture-negative. These data were confirmed using bi-directional sequencing. Evaluation of the fully-automated RVNATSP, which is built on the same detection technology as the VRNAT but contains an updated processor enabling complete automation, revealed the two tests to be functionally equivalent. Thus, the RVNATSP is a fully automated sample-to-result test capable of reliable detection of select respiratory viruses directly from clinical specimens in 3.5 hours.
PMID: 20826644 [PubMed - as supplied by publisher]
Real-Time Detection of Influenza A, Influenza B and Respiratory Syncytial Virus A/B in Respiratory Specimens using Nanoparticle Probes.
Jannetto PJ, Buchan BW, Vaughan KA, Ledford JS, Anderson DK, Henley DC, Quigley NB, Ledeboer NA.
Department of Pathology, Medical College of Wisconsin, and Dynacare Laboratories, 9200 West Wisconsin Ave., Milwaukee, WI 53226, USA; Molecular Pathology Laboratory Network, Inc., 250 East Broadway, Maryville, TN 37804.
Abstract
Seasonal epidemics of influenza and respiratory syncytial virus are responsible for significant morbidity and mortality worldwide. Infrequently, novel or re-emergent strains of influenza A have caused rapid, severe global pandemics resulting in millions of fatalities. The ability to efficiently and accurately detect and differentiate respiratory viruses is paramount to effective treatment, infection control, and epidemiological surveillance. We evaluated the ability of two FDA cleared nucleic acid-based tests, the semi-automated Respiratory Virus Nucleic Acid Test (VRNAT) and the fully automated Respiratory Virus Nucleic Acid Test SP (RVNATSP) (Nanosphere Inc., Northbrook, IL) to detect influenza A, influenza B, and RSV A/B from clinical nasopharyngeal swab specimens. Detection of viral RNA in both tests is based on nucleic acid amplification followed by hybridization to capture probes immobilized on a glass slide. A novel technology utilizing gold nanoparticle conjugated probes is utilized to detect the presence of captured target DNA. This microarray-based approach to detection has proven to be more sensitive than traditional culture/DFA method for detecting RSV and influenza viruses in clinical specimens, including the novel 2009 H1N1 strain. Specifically, we report 98.0% sensitivity and 96.5% specificity of the VRNAT compared to culture/DFA. Further, the VRNAT detected virus in an additional 58% of specimens that were culture-negative. These data were confirmed using bi-directional sequencing. Evaluation of the fully-automated RVNATSP, which is built on the same detection technology as the VRNAT but contains an updated processor enabling complete automation, revealed the two tests to be functionally equivalent. Thus, the RVNATSP is a fully automated sample-to-result test capable of reliable detection of select respiratory viruses directly from clinical specimens in 3.5 hours.
PMID: 20826644 [PubMed - as supplied by publisher]
