title: A comparison of Direct sequencing, Pyrosequencing, High resolution melting analysis, TheraScreen DxS, and the K-ras StripAssay for detecting KRAS mutations in non small cell lung carcinomas.
abstract: BACKGROUND
  It is mandatory to confirm the absence of mutations in the KRAS gene before treating metastatic colorectal cancers with epidermal growth factor receptor inhibitors, and similar regulations are being considered for non-small cell lung carcinomas (NSCLC) and other tumor types. Routine diagnosis of KRAS mutations in NSCLC is challenging because of compromised quantity and quality of biological material. Although there are several methods available for detecting mutations in KRAS, there is little comparative data regarding their analytical performance, economic merits, and workflow parameters.
  
  
  METHODS
  We compared the specificity, sensitivity, cost, and working time of five methods using 131 frozen NSCLC tissue samples. We extracted genomic DNA from the samples and compared the performance of Sanger cycle sequencing, Pyrosequencing, High-resolution melting analysis (HRM), and the Conformité Européenne (CE)-marked TheraScreen DxS and K-ras StripAssay kits.
  
  
  CONCLUSIONS
  Our results demonstrate that TheraScreen DxS and the StripAssay, in that order, were most effective at diagnosing mutations in KRAS. However, there were still unsatisfactory disagreements between them for 6.1% of all samples tested. Despite this, our findings are likely to assist molecular biologists in making rational decisions when selecting a reliable, efficient, and cost-effective method for detecting KRAS mutations in heterogeneous clinical tumor samples.
authors.affiliation: Laboratory of Experimental Medicine, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University and University Hospital in Olomouc, Olomouc, Czech Republic.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Jancik
authors.forename: Jiri
authors.initials: J
authors.lastname: Drabek
authors.forename: Jitka
authors.initials: J
authors.lastname: Berkovcova
authors.forename: Yong Zhong
authors.initials: YZ
authors.lastname: Xu
authors.forename: Marcela
authors.initials: M
authors.lastname: Stankova
authors.forename: Jiri
authors.initials: J
authors.lastname: Klein
authors.forename: Vitezslav
authors.initials: V
authors.lastname: Kolek
authors.forename: Josef
authors.initials: J
authors.lastname: Skarda
authors.forename: Tomas
authors.initials: T
authors.lastname: Tichy
authors.forename: Ivona
authors.initials: I
authors.lastname: Grygarkova
authors.forename: Danuta
authors.initials: D
authors.lastname: Radzioch
authors.forename: Marian
authors.initials: M
authors.lastname: Hajduch
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D015179:Colorectal Neoplasms / Q000235:genetics / Q000556:secondary; D003362:Cost-Benefit Analysis; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D006801:Humans; D009154:Mutation; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000191:economics / Q000379:methods; D018631:ras Proteins / Q000235:genetics*
chemical_list: D004273:DNA, Neoplasm; C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins