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How does it work?

• The MammaTyper® kit uses RNA extracted from standard formalin-fixed, paraffin-embedded (FFPE) sample material from clinical routine, and allows to streamline the molecular pathology workflow to give reliable same day results - including ERBB2 (HER2) results.

• The MammaTyper® test is applicable as a primary diagnostic test (biopsy) for all cases of Breast Cancer. It’s also relevant for the analysis of resection specimens and metastases, as a secondary assessment in the case of unclear IHC results, or as a fast test for ERBB2 (HER2) determination. It is also appropriate for use as an alternative to IHC and in situ hybridisation (FISH/CISH).

• Accurate determination of molecular subtypes is one of several key elements for defining the optimal therapy choice for patients who are diagnosed as having Breast Cancer. MammaTyper® helps to improve subtyping by providing an easily replicated and quantitative measurement of marker gene expression. This way, MammaTyper® helps pathologists to overcome limitations of semi-quantitative staining methods. In essence, what’s been created is something that reduces not only the costs to healthcare providers, but just as importantly, reduces stress and the likelihood of an inappropriate treatment pathway for patients.

Method

The MammaTyper® test is optimized for use in every molecular pathology laboratory. MammaTyper® is based on quantitative one step RT-qPCR technology, combining reverse transcription of mRNA and subsequent quantitative PCR of the resulting cDNA. Signal detection is performed in real time by fluorescently-labelled hydrolysis probes. Expression results are normalised against two reference genes. Additionally, a calibrator corrects for inter-run and inter-instrument variations. Besides quantitative and highly reproducible performance data the test kit delivers fast and reliable results by ready-to-use assay mixes. The kit has been validated on six different real- time PCR instruments:

LightCycler®480instrument II
cobasz®480 Analyser
Versant®kPCR AD module
ABI7500Fast system
CFX96 (BIO-RAD)
MX3000P (Agilent)
Other systems are under evaluation*

The MammaTyper® Kit was developed and validated for use with RNA, which was extracted and purified from formalin-fixed and paraffin-embedded (FFPE) Breast Cancer tissue samples using the RNXtract® RNA extraction kit, according to the manufacturer’s instructions for use. Eluates from the RNXtract® kit can be directly used in the MammaTyper® test. With the fast and easy-to-handle RT-qPCR protocol, quantitative relative gene expression data of the four biomarkers can be generated within one day. MammaTyper® results show a high precision and intra-, inter-run reproducibility. A very low inter-site variation was shown in the course of technical validation of the test. Therefore MammaTyper® precisely determines of the Breast Cancer subtype of a patient, the key for therapy decision and prognosis.

Technology and Workflow

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1

Free biopsy

Cut section

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2

FFPE section

Cut section

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3

RNXtract®

Extract RNA

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4

One step system

everything in one tube

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5

Run RP-qpcr

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6

Patient

Report

Results

The MammaTyper® test delivers a precise, quantitative result for each biomarker. Gene expression for each marker is provided as 40-∆∆Cq values. Based on validated cut-off values (clinical thresholds) each marker is subsequently classified as positive or negative. The combination of the four marker results, accurately determines the subtype according to the St Gallen recommendation. The quantitative and reproducible 40-ΔΔCq values, which are obtained by using the MammaTyper® test, could be the basis for further differentiation of subtypes beyond today’s routine results — an important factor for Individualised treatment in the future.