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Clinical Validation of a Multiplex Kit for RAS Mutations in Colorectal Cancer: Results of the RASKET (RAS KEy Testing) Prospective, Multicenter Study
- Source :
- EBioMedicine, EBioMedicine, Vol 2, Iss 4, Pp 317-323 (2015)
- Publication Year :
- 2014
-
Abstract
- Background RAS (KRAS and NRAS) testing is required to predict anti-epidermal growth factor receptor (EGFR) treatment efficacy in metastatic colorectal cancer (CRC). Although direct sequencing (DS) with manual microdissection (MMD) is widely used, a diagnostic kit providing rapid detections of RAS mutations would be clinically beneficial. We evaluated the MEBGENTM RASKET KIT (RASKET KIT), a multiplex assay using PCR-reverse sequence specific oligonucleotide and xMAP® technology to concurrently detect exon 2, 3, and 4 RAS mutations in a short turnaround time (4.5 h/96-specimens). Methods Formalin-fixed paraffin-embedded (FFPE) tissues were obtained from 308 consenting patients with histologically-confirmed CRC at six hospitals in Japan. For the RASKET KIT, we used only 50–100 ng DNA from each FFPE specimen not processed by MMD. The primary endpoint was the concordance rate between RAS mutations identified with the RASKET KIT and two reference assays (DS with MMD and TheraScreen® K-RAS Mutation Kit). As the secondary endpoints, we evaluated the concordance rate between DS and the RASKET KIT for RAS mutations in the wild-type KRAS exon 2 population and the genotyping performance of the RASKET KIT compared with DS. Findings Among 307 analyzable specimens, the reference assays detected 140 (45.6%, 140/307) RAS mutations: 111 KRAS exon 2 and 29 other (minor) RAS mutations. The RASKET KIT detected 143 (46.6%, 143/307) mutations: 114 KRAS exon 2 and 29 minor RAS mutations. The between-method concordance rate was 96.7% (297/307) (95% CI: 94.1–98.4%). Minor RAS mutations were detected in 15.7% (30/191) of the wild-type KRAS exon 2 population (n = 191); the concordance rate was 98.4% (188/191) (95% CI: 95.5–99.7%). The concordance rate of RAS genotyping was 100% (139/139) (95% CI: 97–100%). Interpretation The RASKET KIT provides rapid and precise detections of RAS mutations and consequently, quicker and more effective anti-EGFR therapy for CRC (Study ID: UMIN000011784). Funding Medical & Biological Laboratories Co., Ltd. (MBL). MBL had roles in study design, data collection, data analysis, and writing of the report for the study.<br />Highlights • We performed clinical evaluation of MEBGEN RASKET KIT to detect RAS mutations. • MEBGEN RASKET KIT detects 48 RAS mutations in a single well. • The overall agreement between MEBGEN RASKET KIT and the gold standard methods was 96.7%. • MEBGEN RASKET KIT provides RAS testing of 96 samples simultaneously. • Results are obtained within 4.5 h per batch using MEBGEN RASKET KIT.
- Subjects :
- Oncology
Neuroblastoma RAS viral oncogene homolog
medicine.medical_specialty
Mutation rate
Genotyping Techniques
Endpoint Determination
Concordance
Population
DNA Mutational Analysis
lcsh:Medicine
Biology
Anti-EGFR antibody treatment
medicine.disease_cause
General Biochemistry, Genetics and Molecular Biology
RAS mutation
Proto-Oncogene Proteins p21(ras)
Mutation Rate
Internal medicine
Genotype
medicine
Humans
Prospective Studies
education
Genotyping
lcsh:R5-920
education.field_of_study
RASKET study
lcsh:R
General Medicine
Exons
Biomarker
Reference Standards
Molecular biology
Colorectal cancer
Mutation
Original Article
KRAS
Reagent Kits, Diagnostic
lcsh:Medicine (General)
Colorectal Neoplasms
In vitro diagnostics
Subjects
Details
- ISSN :
- 23523964
- Volume :
- 2
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- EBioMedicine
- Accession number :
- edsair.doi.dedup.....a261394a685179acf56e323c944d4385