Patient Testing BELAC Accredited

CellCarta’s facility in Belgium received the ISO15189 certification from BELAC to perform numerous diagnostic tests presented below. To order one of these tests, complete the Oncology Request Form (pdf). The form, completed and signed by the requesting pathologist, and the specimens should be sent to the following address:

CellCarta
Sint-Bavostraat 78-80
2610 Wilrijk (Antwerpen)
Belgium

Please consult the following tools to ensure proper test ordering:
Instructions for Completion of the Oncology Request Form
Sample Collection, Handling and Logistics Manual (including shipment and storage)
Additional Information

Results available on our Customer Portal (link)

Contact information:
The lab can be contacted from Monday to Friday between 9h00 and 17h30 CET.
Phone: +32 (0)3 502 0514 or +32 (0)3 502 0519
E-mail: Diagnostics@histogenex.com

ALK Status (Lung Cancer)

To determine the ALK status in non-small-cell lung cancer (NSCLC), a validated ALK IHC test is used as a first screening method. Patient samples that show ALK IHC positivity are subsequently subjected to ALK FISH analysis for confirmation of a positive ALK status.

ALK IHC testing is performed using the Ventana anti-ALK (D5F3) Rabbit monoclonal primary antibody (Roche, CE-IVD) used in combination with the OptiView DAB IHC Detection kit and OptiView Amplification Kit as a fully automated immunohistochemistry assay on the Ventana BenchMark XT automated slide stainer.

ALK FISH testing is performed using the Vysis ALK Break Apart Probe kit (2p23/ALK translocation detection, Abbott, CE-IVD).

ALK Status

BRAF Mutation Testing (Malignant Melanoma, Colorectal Cancer)

The Cobas® 4800 BRAF V600 Mutation Test (Roche Diagnostics) is a real-time PCR based assay designed to detect the presence of the BRAF V600E mutation in formalin-fixed, paraffin-embedded (FFPE) melanoma tissue. The test has been validated at CellCarta to detect BRAF V600 mutations in melanoma and colorectal carcinoma. The test can detect the BRAF V600E mutation at a 5% mutation level using the input of 5 ng/µl DNA. Cross-reactivity is possible with other, non-V600E mutants V600D, V600E2 and V600K.

BRAF Mutation Testing

EGFR Mutation Testing (Lung Cancer)

The Cobas® EGFR Mutation Test v2 in conjunction with the Cobas® 4800 System provides the option of using formalin-fixed, paraffin-embedded (FFPE) tissue or plasma.

The Cobas® EGFR Mutation Test v2 is a real-time PCR test for the qualitative detection and identification of mutations in exons 18, 19, 20 and 21 of the EGFR gene using a DNA input of 50 ng. The test can detect 42 EGFR mutations including G719X (G719A, G719C and G719S) in exon 18; deletions and complex mutations in exon 19; S768I, T790M and insertions in exon 20; L858R and L861Q in exon 21.

In FFPE tumor samples, the presence of mutations is detected with an analytical specificity of at least 95% with a mutation-specific detection limit of at least 5% mutant level in a background of wild-type genomic DNA using the standard input of 50ng per reaction well. In plasma, the presence of mutations is detected with an analytical specificity of at least 95% with a mutation-specific limit of detection of at least 1% mutant level in a background of wild-type genomic DNA using the standard input of 25μL of DNA stock per reaction well.

The Cobas® EGFR Mutation Test v2 is based on two major processes: (1) manual specimen preparation to obtain DNA from FFPE tissues or plasma; (2) PCR amplification and detection of target DNA using complementary primer pairs and oligonucleotide probes labeled with fluorescent dyes. Mutation detection is achieved through PCR analysis with the Cobas z 480 PCR analyzer.

EGFR Mutation Testing

MET Status (Lung Cancer)

The MET fluorescence in situ hybridization (FISH) test used to determine the MET status in non-small-cell lung cancer (NSCLC) is performed using the MET (7q31) / SE7 FISH Probe Kit (Kreatech, RUO).

MET status

MSI Testing (Colorectal Cancer)

OncoMateTM MSI Dx Analysis System (CE-IVD)

The MSI Analysis System is a multiplex, fluorescent PCR-based assay for detection of microsatellite instability. The MSI Analysis System includes fluorescently labelled primers for co-amplification of five quasimonomorphic mononucleotide repeat markers (BAT-25, BAT-26, NR-21, NR-24 and MONO-27). The PCR products are separated by capillary electrophoresis (CE) using an Applied Biosystems 3500Dx Genetic Analyzer. Microsatellite instability at 2 or more mononucleotide loci is interpreted as MSI-high; microsatellite instability at a single mononucleotide locus or no instability at any of the loci tested is interpreted as microsatellite stable (MSS). Typically, MSI-high samples show instability at all loci with an average decrease in product size of 5 to 9 bases. The presence of MSI is detected with an analytical specificity of at least 99% and a detection limit of at least 5%.

An important design feature of the OncoMateTM MSI Dx Analysis System is the spacing between the five different loci, which generally exceeds the typical number of bases shifted (average 5 to 9 bases) when MSI is present.

This assay is the standard for high throughput routine MSI testing, especially when other downstream molecular assays are requested on the same DNA sample.

OncoMate MSI Dx Analysis System (CE-IVD)

IdyllaTM MSI Assay (CE-IVD)

The IdyllaTM MSI Assay performs the detection of microsatellite instability directly from formalin-fixed paraffin embedded (FFPE) human cancer tissue sections utilizing a PCR reaction followed by high-resolution melting curve analysis on an IdyllaTM instrument.

The assay performs detection of mutations in 7 monomorphic homopolymer loci for identification of human cancers with microsatellite instability (ACVR2A, BTBD7, DIDO1, MRE11, RYR3, SEC31A, and SULF2). These biomarkers are tumor-specific, show a high frequency in colorectal and endometrial cancers and are stable across different ethnicities. In addition, these tumor-specific biomarkers do not require the analysis of paired normal tissue samples.

The MSI status for a sample is determined with the final call for all 7 biomarkers. Microsatellite instability at 2 or more mononucleotide loci is interpreted as MSI-high; microsatellite instability at a single mononucleotide locus or no instability at any of the loci tested is interpreted as microsatellite stable (MSS).

This assay allows low throughput MSI testing and is ideal for reflex or rescue testing to rapidly obtain or confirm an accurate MSI result.

Idylla MSI Assay (CE-IVD)

Multi-Gene Mutation and Fusion Detection (Non-Small Cell Lung Cancer - NSCLC)

The AmoyDx®Pan Lung Cancer real-time PCR assay (AmoyDx) in conjunction with the Quantstudio5 System provides the option of using FFPE tissue for fast and accurate mutation and fusion detection.
The Pan Lung Cancer PCR assay is a real time PCR assay for qualitative detection of 167 hotspot alterations in 11 target genes, being EGFR, ALK, ROS1, KRAS, BRAF, HER2, RET, MET, NTRK1, NTRK2 and NTRK3, using a DNA input of 70 ng and an RNA input of 120 ng.

In formalin-fixed, paraffin-embedded tumor samples, the presence of mutations is detected with a mutation-specific sensitivity of 1~5% gene mutation in 7.5-15 ng FFPE tissue DNA. For RNA, the kit allows detection of fusions from 4ng RNA input from FFPE tissue.

Multi-gene mutation and fusion detection in NSCLC

Mutation Testing (Colorectal Cancer)

Cobas® KRAS Mutation Test for In Vitro Diagnostic use (Roche)
The Cobas® KRAS Mutation Test is a real-time PCR test for the qualitative detection of somatic mutations in exon 2 (codons 12/13) and exon 3 (codon 61) of the KRAS gene using a DNA input of 100 ng. The test can detect 19 KRAS mutations. The presence of mutations is detected with an analytical specificity of at least 99% and a detection limit of at least 5% mutant level in a background of wild-type genomic DNA.

KRAS Mutation Test v2 (LSR)
The KRAS Mutation Test v2 (LSR) is an allele-specific, real-time PCR test for the qualitative detection and identification of exon 2, 3, and 4 mutations in the KRAS gene from formalin-fixed, paraffin-embedded (FFPE) tissue. The test is designed to detect 28 unique mutations. The presence of mutations is detected with an analytical specificity of at least 99% and a detection limit of at least 1% mutant level in a background of wild-type genomic DNA.

BRAF/NRAS Mutation Test (LSR)
The BRAF/NRAS Mutation Test (LSR) is an allele-specific, real-time PCR test for the qualitative detection and identification of exon 11 and 15 mutations in the proto-oncogene B-Raf (BRAF) gene and exon 2, 3, and 4 mutations in the neuroblastoma RAS viral oncogene homolog (NRAS) gene from FFPE tissue. The test is designed to detect 36 unique mutations. The presence of mutations is detected with an analytical specificity of at least 99% and a detection limit of at least 5% mutant level in a background of wild-type genomic DNA.

Colorectal Cancer Mutation Testing

PD-L1 (22C3) Status (Lung Cancer)

To determine the PD-L1 status in non-small cell lung cancer (NSCLC), the PharmDx PD-L1 22C3 IHC test from Agilent is used. This Dako 22C3 PharmDx IHC assay is performed as a fully automated immunohistochemistry assay on the Dako autostainer Link 48 automated slide stainer.

PD-L1 (22C3) status

ROS1 Status (Lung Cancer)

ROS1 status in non-small-cell lung cancer (NSCLC) is determine using a validated ROS1 IHC test as a first screening method. Patient samples that show ROS1 IHC overexpression in the tumour cells are subsequently subjected to ROS1 FISH analysis for confirmation of a positive ROS1 status.

ROS1 IHC testing is performed using the Cell Signalling Technology anti-ROS1 (D4D6) Rabbit monoclonal primary antibody used in combination with the OptiView DAB IHC Detection kit and OptiView Amplification Kit as a fully automated immunohistochemistry assay on the Ventana BenchMark Ultra automated slide stainer.

ROS1 FISH testing is performed using the Vysis 6q22 ROS1 Break apart FISH probe (Abbott, RUO).

ROS1 Status