Design and performance testing of quantitative real time PCR assays for influenza A and B viral load measurement.

摘要: Background: The antiviral effect of anti-influenza drugs such as zanamivir may be demonstrated in patients an increased rate decline viral load over a time course treatment compared with placebo. Historically this was measured using plaque assays, or Culture Enhanced Enzyme Linked Immunosorbent Assay (CE-ELISA). Objectives: to develop and characterise real quantitative PCR (qPCR) assays measure influenza A B clinical samples, that offer improvements existing methods, particular virus infectivity assays. Study design: dynamic range robustness were established for the qPCR along stability assay components. Cross validation CE-ELISA performed by parallel testing both serial dilutions three different subtypes cultured panel positive throat swab specimens. Results: specific ranges at least seven logs. variability within acceptable limits but towards lower limit quantification, which 3.33 log10 cDNA copies/ml transport medium (ten RNA copies/PCR). components robust enough withstand extended storage several freeze–thaw cycles. For quantification equivalent cut off, equates 93-fold increase sensitivity. Conclusion: Well characterised significant methods measuring strains