title: The mutation rates of EGFR in non-small cell lung cancer and KRAS in colorectal cancer of Chinese patients as detected by pyrosequencing using a novel dispensation order.
abstract: BACKGROUND
  The purpose of this study was to develop a cost-effective approach for the determination of EGFR and KRAS mutations in formalin-fixed paraffin-embedded (FFPE) non-small cell lung cancer (NSCLC) and colorectal cancer (CRC) samples from Chinese patients based on a sensitive pyrosequencing (PS) technique.
  
  
  METHODS
  The NSCLC and CRC cell lines were tested to determine the limitation of detection and reproducibility of the PS method. In addition, 494 NSCLC and 1099 CRC patient samples were assayed by PS to evaluate the EGFR or KRAS mutation patterns according to the clinicopathological features.
  
  
  RESULTS
  The PS assay was able to reproducibly detect as few as 2 % mutant alleles with excellent linearity. EGFR mutations were detected in 35.63 % of the NSCLC samples, and KRAS mutations were detected in 39.76 % of the CRC samples. EGFR mutations were more frequently observed to be significant by multivariate analysis in NSCLC patients who were 65 years old or younger (OR = 2.51), had a nonsmoking history (OR = 3.63), and adenocarcinoma (OR = 3.57), but not in females (OR = 0.64). KRAS mutations were more frequently detected in CRC patients who were female (OR = 1.64) and 50 years old or older (OR = 4.17), and had adenocarcinoma (OR = 2.41).
  
  
  CONCLUSIONS
  This is the first extensive validation of PS on FFPE samples using the detection of EGFR exons 18-21 mutations and KRAS exon 2 mutations. Our results demonstrate the utility of PS analysis for the detection of somatic EGFR and KRAS mutations in clinical samples and provide important clinical and molecular characteristics of NSCLC and CRC from Chinese patients.
authors.affiliation: Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kong Jiang Road, Shanghai, 200092, China. xghsent@163.com.
authors.forename: Guohua
authors.initials: G
authors.lastname: Xie
authors.affiliation: Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kong Jiang Road, Shanghai, 200092, China. pepper_xief@126.com.
authors.forename: Fang
authors.initials: F
authors.lastname: Xie
authors.affiliation: Department of General Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200092, China. wu_ping126@163.com.
authors.forename: Ping
authors.initials: P
authors.lastname: Wu
authors.affiliation: Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kong Jiang Road, Shanghai, 200092, China. yuanxiangliang@gmail.com.
authors.forename: Xiangliang
authors.initials: X
authors.lastname: Yuan
authors.affiliation: Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kong Jiang Road, Shanghai, 200092, China. 13524699449@126.com.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kong Jiang Road, Shanghai, 200092, China. freddy840924@hotmail.com.
authors.forename: Yunchuan
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kong Jiang Road, Shanghai, 200092, China. lily107113@163.com.
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kong Jiang Road, Shanghai, 200092, China. celine_xu@live.cn.
authors.forename: Ling
authors.initials: L
authors.lastname: Xu
authors.affiliation: Department of Anorectal Surgery and Colorectal Cancer Center, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kong Jiang Road, Shanghai, 200092, China. yangming_xh@163.com.
authors.forename: Ming
authors.initials: M
authors.lastname: Yang
authors.affiliation: Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kong Jiang Road, Shanghai, 200092, China. lisongshen@yeah.net.
authors.forename: Lisong
authors.initials: L
authors.lastname: Shen
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D044466:Asian People; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D059645:Mutation Rate; D017422:Sequence Analysis, DNA / Q000379:methods*; D018631:ras Proteins / Q000235:genetics*
chemical_list: D066246:ErbB Receptors; D018631:ras Proteins