title: A novel, rapid point-of-care test for lung cancer patients to detect epidermal growth factor receptor gene mutations by using real-time droplet-PCR and fresh liquid cytology specimens.
abstract: Epidermal growth factor receptor gene (EGFR) mutations are associated with response to tyrosine kinase inhibitors (TKIs) in patients with non-small cell lung cancer (NSCLC). We developed a novel, rapid EGFR mutation assay using a real-time droplet-polymerase chain reaction machine (EGFR d-PCR assay). The purpose of this study was to validate the performance of the EGFR d-PCR assay using fresh liquid cytology specimens. We analyzed three major EGFR mutations (L858R in exon 21, E746_A750del in exon 19 and T790M in exon 20) in 80 fresh liquid cytology specimens of adenocarcinoma (ADC) or NSCLC-not otherwise specified (NOS) via the EGFR d-PCR assay and conventional real-time PCR assay using the therascreen® EGFR RGQ PCR kit (Therascreen assay). In addition, we performed sensitivity assays using cell lines with EGFR mutations. The EGFR d-PCR assay detected 16 L858Rs, 8 E746_A750dels and 1 T790M mutation and the Therascreen assay detected 16 L858Rs, 11 deletions in exon 19 and 1 T790M mutation. The results were concordant between the two assays. The reaction time of the EGFR d-PCR assay was 8 min and 10 sec, but that of the Therascreen assay was 1 h and 45 min. Sensitivity, as assessed by the detection limit of the EGFR d-PCR assay was 0.5, 0.05 and 0.5% for L858R, E746_A750del and T790M, respectively. The EGFR d-PCR assay markedly reduced the detection time of major EGFR mutations with high sensitivity compared with the conventional Therascreen assay and is expected to expedite EGFR-TKI therapy for lung cancer patients, especially those in advanced stages.
authors.affiliation: Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Shiho
authors.initials: S
authors.lastname: Asaka
authors.affiliation: Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Yoshizawa
authors.affiliation: Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Matsuda
authors.affiliation: Graduate School of Science and Technology, Shinshu University, Wakasato, Nagano 380-8553, Japan.
authors.forename: Akemi
authors.initials: A
authors.lastname: Yamaguchi
authors.affiliation: First Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamamoto
authors.affiliation: Department of Thoracic Surgery, Shinshu University Hospital, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Shiina
authors.affiliation: Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Nakata
authors.affiliation: Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Ogawa
authors.affiliation: Department of Thoracic Surgery, Shinshu University Hospital, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Meng
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Nagano 390-8621, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Honda
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology; D000077192:Adenocarcinoma of Lung; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D044963:Biopsy, Fine-Needle; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics / Q000473:pathology; D004252:DNA Mutational Analysis / Q000379:methods*; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D010996:Pleural Effusion / Q000235:genetics / Q000473:pathology; D019095:Point-of-Care Systems; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: C512478:EGFR protein, human; D066246:ErbB Receptors