title: Detection of Epidermal Growth Factor Receptor Mutations in Lung Adenocarcinoma: Comparing Cobas 4800 EGFR Assay With Sanger Bidirectional Sequencing.
abstract: Accurate detection of epidermal growth factor receptor (EGFR) mutations has a crucial role in the current treatment of patients with lung adenocarcinoma, and identification of clinically relevant mutations would qualify patients for treatment with tyrosine kinase inhibitors. Historically, Sanger sequencing has been used as the reference standard assay for EGFR mutational analysis; however, Cobas 4800 is a relatively new method. In the present study, we compared the performance of the Cobas assay against that of Sanger sequencing.
  
  
  
  A total of 493 consecutive formalin-fixed paraffin-embedded samples of lung adenocarcinoma were simultaneously tested for EGFR mutations using both methods.
  
  
  
  After exclusion of the invalid results (n = 19), 474 samples from 455 patients were analyzed. The Cobas assay showed a mutation detection rate comparable to that of Sanger sequencing (18.1% vs. 17.9%, respectively; P < .05). Excellent agreement of 98.9% (κ, 0.964) was observed between the 2 methods.
  
  
  
  The Cobas assay is a fast and diagnostically robust platform with high analytical sensitivity; however, it is limited by its detection range and low tolerance to low DNA quality. Sanger sequencing is mostly affected by its lower analytic sensitivity. Ultimately, a dual testing strategy will be justified to increase the detection of novel mutations and reduce the false-negative results within an acceptable turnaround time.
authors.affiliation: Department of Anatomical Pathology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Perth, Western Australia, Australia. Electronic address: nima.mesbahardakani@health.wa.gov.au.
authors.forename: Nima Mesbah
authors.initials: NM
authors.lastname: Ardakani
authors.affiliation: Department of Anatomical Pathology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Perth, Western Australia, Australia.
authors.forename: Tindaro
authors.initials: T
authors.lastname: Giardina
authors.affiliation: Department of Anatomical Pathology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Perth, Western Australia, Australia.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Grieu-Iacopetta
authors.affiliation: Department of Anatomical Pathology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Perth, Western Australia, Australia.
authors.forename: Yordanos
authors.initials: Y
authors.lastname: Tesfai
authors.affiliation: Department of Anatomical Pathology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Perth, Western Australia, Australia.
authors.forename: Amerigo
authors.initials: A
authors.lastname: Carrello
authors.affiliation: Department of Anatomical Pathology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Perth, Western Australia, Australia.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Taylor
authors.affiliation: Department of Anatomical Pathology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Perth, Western Australia, Australia.
authors.forename: Cleo
authors.initials: C
authors.lastname: Robinson
authors.affiliation: Department of Anatomical Pathology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Perth, Western Australia, Australia; University of Western Australia, School of Pathology and Laboratory Medicine, Perth, Western Australia, Australia.
authors.forename: Dominic
authors.initials: D
authors.lastname: Spagnolo
authors.affiliation: Department of Anatomical Pathology, PathWest Laboratory Medicine, Queen Elizabeth II Medical Centre, Perth, Western Australia, Australia; University of Western Australia, School of Pathology and Laboratory Medicine, Perth, Western Australia, Australia.
authors.forename: Benhur
authors.initials: B
authors.lastname: Amanuel
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics*; D000077192:Adenocarcinoma of Lung; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D066246:ErbB Receptors / Q000235:genetics*; D005188:False Negative Reactions; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*; D013997:Time Factors
chemical_list: D066246:ErbB Receptors
keywords: Cobas assay; EGFR mutations; Lung adenocarcinoma; Sanger sequencing; Tyrosine kinase inhibitor