The Easy® kits allow the qualitative detection of the main somatic mutations of the genes EGFR, KRAS, NRAS and BRAF by Real-Time PCR in association with a system of enrichment of the mutated allele.

Each kit includes all the reagents necessary for the test and the positive controls of reaction.

All the kits share the same thermal profile for the detection of the somatic mutations.

Main features Detection of the main mutations of exon 2 (codons 12, 13), of exon 3 (codons 59, 61) and of exon 4 (codons 117, 146) of the gene KRAS using 12 oligo mixes. Each mix allows the co-amplification of one or more mutated alleles plus an endogenous control gene. A specifc oligo control mix enables the evaluation of the quality and the quantity of the DNA in each sample.

Controls Control DNA positive for all the mutations detected by the kit. Reference standard DNA Horizon KRAS G12V 1% to monitor the analytical process and the performances of the system. The standard is characterized by a well defined allelic ratio wild-type/mutant.

Sensitivity The kit allows the detection of low percentages of mutated allele in presence of high amounts of wildtype genomic DNA by real-time amplification with sequence-specific probes marked with FAM and HEX (LOD down to 0.5%).

Starting material The kit allows the analysis of DNA extracted from fresh, frozen and formalin-fixed paraffin-embedded tissue.

Execution time  2 hours.

Main features Detection of the main mutations of codon 600 of the gene BRAF using 5 oligo mixes. Each mix allows the co-amplification of one or more mutated alleles plus an endogenous control gene. A specific oligo control mix enables the evaluation of the quality and the quantity of the DNA in each sample. Control DNA positive for all the mutations detected by the kit.

Controls Reference standard DNA Horizon BRAF V600E 1% to monitor the analytical process and the performances of the system. The standard is characterized by a well defined allelic ratio wild-type/mutant.

Sensitivity The kit allows the detection of low percentages of mutated allele in presence of high amounts of wild-type genomic DNA by real-time amplification with sequence-specific probes marked with FAM and HEX (LOD down to 0.5%).

Starting material The kit allows the analysis of DNA extracted from fresh, frozen and formalin-fixed paraffin-embedded tissue.

Execution time  2 hours.

Main features Detection of the main mutations of exons 18, 19, 20, 21 of EGFR gene using 8 oligo mixes. Each mix allows the co-amplification of one or more mutated alleles plus an endogenous control gene. A specific oligo control mix enables the evaluation of the quality and the quantity of the DNA in each sample.

Controls Control DNA positive for all the mutations detected by the kit. Reference standard DNA Horizon EGFR E746-A750 1% to monitor the analytical process and the performances of the system. The standard is characterized by a well defined allelic ratio wild-type/mutant.

Sensitivity The kit allows the detection of low percentages of mutated allele in presence of high amounts of wild-type genomic DNA by real-time amplification with sequence-specific probes marked with FAM and HEX (LOD down to 0.5%).

Starting material The kit allows the analysis of DNA extracted from fresh, frozen and formalin-fixed paraffin-embedded tissue.

Execution time  2 hours.

Main features Detection of the main mutations of exon 2 ( codons 12, 13), of exon 3 (codons 59, 61) and of exon 4 (codons 117, 146) of NRAS gene of EGFR gene using 8 oligo mixes. Each mix allows the co-amplification of one or more mutated alleles plus an endogenous control gene. A specific oligo control mix enables the evaluation of the quality and the quantity of the DNA in each sample.

Controls Control DNA positive for all the mutations detected by the kit. Reference standard DNA Horizon NRAS Q61K 1% to monitor the analytical process and the performances of the system. The standard is characterized by a well defined allelic ratio wild-type/mutant.

Sensitivity The kit allows the detection of low percentages of mutated allele in presence of high amounts of wild-type genomic DNA by real-time amplification with sequence-specific probes marked with FAM and HEX (LOD down to 0.5%).

Starting material The kit allows the analysis of DNA extracted from fresh, frozen and formalin-fixed paraffin-embedded tissue.

Execution time  2 hours.

Main features Qualitative detection of aberrant expression of the tyrosin-kinase domain of ALK associated to the gene fusion by retrotrascription and co-amplification of a region located at 3’ terminus of ALK mRNA (detected by a FAM marked probe) and of an endogenous control gene (detected by a HEX marked probe).

Controls Control RNA for the monitoring of the analytical process.

Sensitivity The kit allows the detection of low percentages of mutated allele in presence of high amounts of wild-type genomic DNA by real-time amplification with sequence-specific probes marked with FAM and HEX (LOD down to 0.5%).

Starting material The kit allows the analysis of DNA extracted from fresh, frozen and formalin-fixed paraffin-embedded tissue.

Execution time  2 hours.

Main features Detection, by allelic discrimination, of the DPYD gene polymorphisms (IVS10C>G, HapB3, c.1236G>A), DPYD*13 (c.1679T>G), DPYD*2A (IVS14+1G>A, c.1905+1G>A), DPYD c.2846>T, DPYD c.2194G>A, associated with the toxicity due to the treatment with fluoropyrimidines, using 4 oligo mixes. Each mix allows the co-amplification of the mutant sequence (FAM) as well as the wild-type sequence (HEX).

Controls DPYD WT positive control: Positive control DNA containing a mixture of synthetic wild-type DNA sequences for DPYD polymorphisms analyzed. DPYD MT positive control: Positive control DNA containing a mixture of synthetic mutant DNA sequences for DPYD polymorphisms analyzed.

Starting material The kit allows the analysis of genomic DNA extracted from whole blood.

Execution time  2 hours.

Main features Detection, by allelic discrimination, of the UGT1A1 gene polymorphisms UGT1A1*1 (TA)6, UGT1A1*28 (TA)7, UGT1A1*36 (TA)5 and UGT1A1*37 (TA)8, associated with the toxicity due to the treatment with irinotecan, using 1 oligo mix. UGT1A1 mix contains HEX labeled probes for UGT1A1*28 and UGT1A1*37 and FAM labeled probes for UGT1A1*1 and UGT1A1*36.

Controls UGT1A1 WT positive control: Positive control DNA containing synthetic wild-type UGT1A1*1/*1 DNA sequence. UGT1A1 MT positive control: Positive control DNA containing synthetic mutant UGT1A1*28/*28 DNA sequence.

Starting material The kit allows the analysis of genomic DNA extracted from whole blood.

Execution time  2 hours.

Main features Detection of the main mutations of exon 2 (codons 12,13), of exon 3 (codons 61) of the genes KRAS, NRAS, HRAS and of the codons 600 and 601 of the gene BRAF using 8 oligo mixes. Each mix allows the co-amplification of one or more mutated alleles plus an endogenous control gene.

Controls Controls Positive control DNA containing a mixture of synthetic DNA sequences that correspond to each mutation detected by this kit in a background of wild-type genomic DNA.

Starting material The kit allows the analysis of genomic DNA extracted from fresh, frozen or formalin fixed paran-embedded (FFPE) tumor tissue. The kit allows the analysis of genomic DNA extracted from cytological samples.

Execution time  2 hours.

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    Category

    Realtime PCR

    Validated tools
    Rotor-Gene Q
    Rotor-Gene 6000
    Stratagene MX3000P
    Stratagene MX3005P
    ABI 7300
    ABI 7500
    Bio-Rad CFX96
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