Performance Data
Detection of Colorectal Cancer-Associated BRAF V600E Mutation from 15 mg/mL Human Stool Lyo-Ready Genotyping Direct qPCR Stool combines the latest advances in buffer chemistry and PCR enhancers, together with an optimized antibody-mediated hot-start polymerase, for fast, precise, and highly reproducible allelic discrimination and cluster separation with SNP detection assays, even in the presence of PCR inhibitors found in stool.
A)
B)
Lyo-Ready
KAPA
Detection of colorectal cancer-associated BRAF V600E mutation (SNP accession number: rs113488022) using A) Lyo-Ready ™ Genotyping Direct qPCR Stool (MDX148), B) KAPA Probe Force qPCR Mix (Roche), C) TaqPath ™ ProAmp ™ Multiplex Master Mix (ThermoFisher) and D) Type-it Fast SNP Probe PCR Kits (Qiagen) in presence of 15 mg/mL human stool. BRAF V600E (val600-to-glu) mutation is a prognostic and predictive biomarker of aggressive tumor growth and treatment with EGFR inhibitors. BRAF testing is recommended for stage IV, metastatic colorectal cancer (mCRC) before initiating therapy with anti-EGFR inhibitors, or when chemotherapy treatment stops working 1 . The results illustrate the sensitivity of Lyo-Ready ™ Genotyping Direct qPCR Stool to detect the presence of low levels of clinically relevant mutations directly from stool samples.
Mutant amplification
WT
C)
D)
TaqPath
Type-it
Detection of Gastric Cancer-Associated p53 Mutation from 20 mg/mL Human Stool
A)
B)
Detection of colorectal cancer-associated p53 mutation (SNP accession number: rs1042522) was performed using A) Lyo-Ready ™ Genotyping Direct qPCR Stool, B) Kapa Probe Force, C) TaqPath ™ and D) Type-it Fast Kits, in presence of 20 mg/mL human stool. Homozygous Allele C ( red ) and Allele G ( blue ) and heterozygous Allele C/G ( green ) with a NTC ( black ) and x for undetermined. Approximately half of all colorectal cancers show p53 gene mutations, and advanced gastric cancer treated with paclitaxel and cisplatin combination chemotherapy found that rs1042522(G;G) and (C;G) genotypes correlate with worse progression 2 . The results illustrate the ability of Lyo-Ready ™ Genotyping Direct qPCR Stool to form tight clusters and to achieve accurate allelic discrimination of clinically relevant mutations directly from crude human stool samples.
C)
D)
Allele G | Allele C | Allele C/G | NTC
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