title: Considerations for accurate gene expression measurement by reverse transcription quantitative PCR when analysing clinical samples.
abstract: Reverse transcription quantitative PCR is an established, simple and effective method for RNA measurement. However, technical standardisation challenges combined with frequent insufficient experimental detail render replication of many published findings challenging. Consequently, without adequate consideration of experimental standardisation, such findings may be sufficient for a given publication but cannot be translated to wider clinical application. This article builds on earlier standardisation work and the MIQE guidelines, discussing processes that need consideration for accurate, reproducible analysis when dealing with patient samples. By applying considerations common to the science of measurement (metrology), one can maximise the impact of gene expression studies, increasing the likelihood of their translation to clinical tools.
authors.affiliation: Molecular & Cell Biology, LGC, Queens Road, Teddington, TW11 0LY, UK, rebecca.sanders@lgcgroup.com.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Sanders
authors.forename: Deborah J
authors.initials: DJ
authors.lastname: Mason
authors.forename: Carole A
authors.initials: CA
authors.lastname: Foy
authors.forename: Jim F
authors.initials: JF
authors.lastname: Huggett
mesh_terms: D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards; D005820:Genetic Testing / Q000379:methods / Q000592:standards; D006801:Humans; D012313:RNA / Q000032:analysis / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D048348:Reverse Transcription
chemical_list: D012313:RNA