title: PCR-sequencing is a complementary method to amplification refractory mutation system for EGFR gene mutation analysis in FFPE samples.
abstract: Amplification Refractory Mutation System (ARMS) is the most popular technology for EGFR gene mutation analysis in China. Cutoff Ct or ΔCt values were used to differentiate low mutation abundance cases from no mutation cases. In this study, all of 359 NSCLC samples were tested by ARMS. Seventeen samples with larger Ct or ΔCt than cutoff values were retested by PCR-sequencing. TKI treatment responses were monitored on the cases with ARMS negative and PCR-sequencing positive results. One exon 18 G719X case, 67 exon 19 deletion cases, 2 exon 20 insertion cases, 1 exon 20 T790M case, 60 exon 21 L858R cases, 5 exon 21 L861Q cases and 201 wild type cases were identified by ARMS. Another 22 cases were evaluated as wild type but had later amplification fluorescent curves. Seventeen out of these 22 cases were retested by PCR-sequencing. It turns out that 3 out of 3 cases with exon 19 deletion later amplifications, 2 out of 2 cases with L858R later amplifications and 4 out of 12 cases with T790M later amplifications were identified as mutation positive. Two cases with exon 19 deletion and L858R respectively were treated by TKI and got responses. Our study indicated that PCR-sequencing might be a complementary way to confirm ARMS results with later amplifications.
authors.affiliation: Department of Pathology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China.
authors.forename: Junchang
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Department of Pathology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China; Department of Pathology, The Second Hospital of Jiaxing, Jiaxing, Zhejiang 314000, China.
authors.forename: Chunhua
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Pathology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Yu
authors.affiliation: Department of Pathology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China.
authors.forename: Danli
authors.initials: D
authors.lastname: Sheng
authors.affiliation: Department of Pathology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China; Key Laboratory of Diagnostic Medicine designated by the Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Zuo
authors.affiliation: Department of Pathology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China.
authors.forename: Minpu
authors.initials: M
authors.lastname: Ren
authors.affiliation: Department of Pathology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China.
authors.forename: Yaqin
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Ophthalmology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Shen
authors.affiliation: Department of Pathology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China.
authors.forename: Mei
authors.initials: M
authors.lastname: Jin
authors.affiliation: Department of Pathology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China. Electronic address: xusx1979@hotmail.com.
authors.forename: Songxiao
authors.initials: S
authors.lastname: Xu
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D005557:Formaldehyde; D018773:Genes, erbB-1 / Q000235:genetics*; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D016707:Tissue Fixation
chemical_list: D005557:Formaldehyde
keywords: ARMS; Complementary method; EGFR mutation; PCR-sequencing