FGFR RGQ RT-PCR Kit

• Detection of four point mutations and five fusions
• High sensitivity and specificity
• Simple workflow with next-day results
• Automated data analysis using Rotor-Gene AssayManager v2.1 software

The FGFR RGQ RT-PCR Kit is a qualitative assay for the detection of four point mutations in of the FGFR3 gene: exon 7 [p.R248C (c.742C>T), p.S249C (c.746C>G)] and exon 10 [p.G370C (c.1108G>T), p.Y373C (c.1118A>G)]. In addition, the assay detects three fusions in the FGFR3 gene (FGFR3-TACC3v1, FGFR3-TACC3v3 and FGFR3-BAIAP2L1) and two fusions in the FGFR2 gene (FGFR2-BICC1 and FGFR2-CASP7).

The FGFR RGQ RT-PCR Kit is based on the selective amplification of alterations in the FGFR2 and FGFR3 genes using the Rotor-Gene Q 5plex HRM instrument for sensitive and specific analysis. The assay exploits the qPCR oligonucleotide hydrolysis principle using TaqMan probes.

Allele-specific technology allows accurate and highly reproducible detection of alterations, based on the use of specific forward and reverse primers and probes; only a perfect match between the primers and probes with the target cDNA allows extension and amplification in the PCR reaction. Data analysis is performed automatically. Outputs for each PCR are displayed individually, with "Amplification Detected" displayed when a specific FGFR mutation is detected.

The fast and simple workflow takes ~12 hours from Sample to Insight.

RNA is first prepared from formalin-fixed paraffin-embedded (FFPE) urothelial tumor samples using the RNeasy DSP FFPE Kit or RNeasy FFPE Kit. Purified RNA is then reverse transcribed using Reverse Transcriptase to generate cDNA for real-time PCR analysis. Optimized reverse transcription reagents and PCR master mixes provided in the kit enable high-fidelity reverse transcription and sensitive real-time PCR on the Rotor-Gene Q 5plex HRM instrument. Qualitative results are displayed in Rotor-Gene AssayManager software, informing the system operator if one or more of the four point mutations and five fusions detected by the kit are present in each sample.

The FGFR RGQ RT-PCR Kit enables qualitative detection of four point mutations and five fusions in the FGFR2 and FGFR3 genes.