title: Analytical Performance of a 15-Gene Prognostic Assay for Early-Stage Non-Small-Cell Lung Carcinoma Using RNA-Stabilized Tissue.
abstract: A 15-gene prognostic signature for early-stage, completely resected, non-small-cell lung carcinoma, (which distinguishes between patients with good and poor prognoses) was clinically validated in prior studies. To achieve operational efficiencies, this study was designed to evaluate the assay's performance in RNA-stabilized tissue as an alternative to the fresh-frozen tissue format originally used to develop the assay. The percent concordance between matched tissue formats was 84% (95% Wilson CI, 70%-92%), a level of agreement comparable to the inherent reproducibility of the assay observed within biological replicates of fresh-frozen tissue. Furthermore, the analytical performance of the assay using the RNA-stabilized tissue format was evaluated. When compared to an accredited reference laboratory, the clinical laboratory achieved a concordance of 94% (95% Wilson CI, 81%-98%), and there was no evidence of bias between the laboratories. The lower limit of quantitation for the target RNA concentration was confirmed to be, at most, 12.5 ng/μL. The assay reportable range defined in terms of risk score units was determined to be -4.295 to 4.210. In a large-scale precision study, the assay showed high reproducibility and repeatability. When subjected to a maximal amount of genomic DNA, a potential contaminant, the assay still produced the expected results. The 15-gene signature was confirmed to produce reliable results and, thus, is suitable for its intended use.
authors.affiliation: Helomics Corporation, Pittsburgh, Pennsylvania.
authors.forename: Shuguang
authors.initials: S
authors.lastname: Huang
authors.affiliation: Helomics Corporation, Pittsburgh, Pennsylvania.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Reitze
authors.affiliation: Helomics Corporation, Pittsburgh, Pennsylvania.
authors.forename: Amy L
authors.initials: AL
authors.lastname: Ewing
authors.affiliation: Helomics Corporation, Pittsburgh, Pennsylvania.
authors.forename: Suzanne
authors.initials: S
authors.lastname: McCreary
authors.affiliation: Helomics Corporation, Pittsburgh, Pennsylvania.
authors.forename: Arlette H
authors.initials: AH
authors.lastname: Uihlein
authors.affiliation: Helomics Corporation, Pittsburgh, Pennsylvania.
authors.forename: Stacey L
authors.initials: SL
authors.lastname: Brower
authors.affiliation: Helomics Corporation, Pittsburgh, Pennsylvania.
authors.forename: Dakun
authors.initials: D
authors.lastname: Wang
authors.affiliation: Helomics Corporation, Pittsburgh, Pennsylvania.
authors.forename: Tianhua
authors.initials: T
authors.lastname: Wang
authors.affiliation: Helomics Corporation, Pittsburgh, Pennsylvania.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Gabrin
authors.affiliation: Almac Diagnostics Ltd., Craigavon, Northern Ireland, United Kingdom.
authors.forename: Katherine E
authors.initials: KE
authors.lastname: Keating
authors.affiliation: Almac Diagnostics Ltd., Craigavon, Northern Ireland, United Kingdom.
authors.forename: Jude
authors.initials: J
authors.lastname: Mulligan
authors.affiliation: Almac Diagnostics Ltd., Craigavon, Northern Ireland, United Kingdom.
authors.forename: Claire
authors.initials: C
authors.lastname: Wilson
authors.affiliation: Almac Diagnostics Ltd., Craigavon, Northern Ireland, United Kingdom.
authors.forename: Timothy
authors.initials: T
authors.lastname: Davison
authors.affiliation: Almac Diagnostics Ltd., Craigavon, Northern Ireland, United Kingdom.
authors.forename: Stuart
authors.initials: S
authors.lastname: McKenzie
authors.affiliation: University Health Network, Princess Margaret Cancer Centre, Toronto, Ontario, Canada.
authors.forename: Ming-Sound
authors.initials: MS
authors.lastname: Tsao
authors.affiliation: University Health Network, Princess Margaret Cancer Centre, Toronto, Ontario, Canada.
authors.forename: Frances A
authors.initials: FA
authors.lastname: Shepherd
authors.affiliation: Helomics Corporation, Pittsburgh, Pennsylvania. Electronic address: vplamadeala@helomics.com.
authors.forename: Victoria
authors.initials: V
authors.lastname: Plamadeala
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics*; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis* / Q000235:genetics / Q000628:therapy; D004273:DNA, Neoplasm / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000628:therapy; D016612:Paraffin Embedding; D011379:Prognosis; D012334:RNA, Neoplasm / Q000737:chemistry*; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D012680:Sensitivity and Specificity
chemical_list: D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D012334:RNA, Neoplasm; D011933:Reagent Kits, Diagnostic