ArticleJournal of clinical virology : the official publication of the Pan American Society for Clinical Virology2004
Design and performance testing of quantitative real time PCR assays for influenza A and B viral load measurement.
Article in Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 155 papers.
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Who cites it
155 citing papers in PubMed.
- Access to a polymerase chain reaction assay method targeting 13 respiratory viruses can reduce antibiotics: a randomised, controlled trial.BMC medicine · 2011Trial
- Human bocavirus and acute wheezing in children.Clinical infectious diseases : an official publication of the Infectious Diseases Society of America · 2007Trial
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- Epidemiology of Infectious Pathogens in Horses with Acute Respiratory Disease, Abortion, and Neurological Signs: Insights Gained from the Veterinary Surveillance System for Horses in The Netherlands (SEIN).Veterinary sciences · 2025Article
- Development and Laboratory Validation of Rapid, Bird-Side Molecular Diagnostic Assays for Avian Influenza Virus Including Panzootic H5Nx.Microorganisms · 2025Article
- Genetic reinstatement of RIG-I in chickens reveals insights into avian immune evolution and influenza interaction.Frontiers in immunology · 2025Article
- Vacuum Oven Drying: A Cost-Effective Way of Producing Field-Deployable Reagents for In-house Real-Time PCR Methods.Molecular biotechnology · 2025Article
- Stability of influenza A virus in droplets and aerosols is heightened by the presence of commensal respiratory bacteria.Journal of virology · 2024Article
- In vitro one-pot construction of influenza viral genomes for virus particle synthesis based on reverse genetics system.PloS one · 2024Article
- Submerged leaves of live indoor foliage plants adsorb H1N1 influenza virus from suspension.Plant signaling & behavior · 2023Article
- Retrospective Analysis of the Detection of Pathogens Associated with the Porcine Respiratory Disease Complex in Routine Diagnostic Samples from Austrian Swine Stocks.Veterinary sciences · 2023Article
95 more citing papers are in PubMed but not listed here.
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Authors and funding
8 authors.
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No grant is acknowledged in the PubMed record.
Abstract
backgroundThe antiviral effect of anti-influenza drugs such as zanamivir may be demonstrated in patients as an increased rate of decline in viral load over a time course of treatment as compared with placebo. Historically this was measured using plaque assays, or Culture Enhanced Enzyme Linked Immunosorbent Assay (CE-ELISA).
objectivesto develop and characterise real time quantitative PCR (qPCR) assays to measure influenza A and B viral load in clinical samples, that offer improvements over existing methods, in particular virus infectivity assays. STUDY
designThe dynamic range and robustness were established for the real time qPCR assays along with stability of the assay components. Cross validation of the real time PCR assays with CE-ELISA was performed by parallel testing of both serial dilutions of three different subtypes of cultured virus and a panel of influenza positive throat swab specimens.
resultsthe assays were specific for influenza A and B and the dynamic ranges were at least seven logs. The assay variability was within acceptable limits but increased towards the lower limit of quantification, which was 3.33 log(10) viral cDNA copies/ml of virus transport medium (ten viral RNA copies/PCR). The components of the assay were robust enough to withstand extended storage and several freeze-thaw cycles. For the real time PCR assays the limit of quantification was equivalent to the virus infectivity cut off, which equates to a 93-fold increase in sensitivity.
conclusionWell characterised real time PCR assays offer significant improvements over the existing methods for measuring the viral load of strains of influenza A and B in clinical specimens.
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