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Table 3 Validation metrics of MammaTyper amplification for each different target sequence and sample type

From: Technical validation of an RT-qPCR in vitro diagnostic test system for the determination of breast cancer molecular subtypes by quantification of ERBB2, ESR1, PGR and MKI67 mRNA levels from formalin-fixed paraffin-embedded breast tumor specimens

Versant kPCR ERBB2 ESR1 PGR MKI67 B2M CALM2
FFPE Master Sample LoD [Cq] 34.36 35.54 32.33 35.66 35.80 34.78
LoD [molecules] 10 5 87 7 7 8
AMR [Cq] 21.05–34.36 19.13–35.54 21.13–32.33 23.97–35.66 16.78–35.80 23.90–34.78
Efficiency [%] 106 105 106 101 100 111
Linearity 0.999 0.999 0.995 0.999 0.999 0.998
IVT-RNA LoD [Cq] 35.31 35.60 36.72 36.41 36.22 33.69
LoD [molecules] 5 5 5 5 5 18
AMR [Cq] 12.36–35.31 12.45–35.60 12.64–36.72 11.63–36.41 12.41–36.22 11.67–33.69
Efficiency [%] 105 104 100 99 100 100
Linearity 0.999 0.999 1.000 0.999 1.000 1.000
LightCycler 480 II ERBB2 ESR1 PGR MKI67 B2M CALM2
FFPE Master Sample LoD [Cq] 33.56 36.47 33.56 35.47 35.6 35.62
LoD [molecules] 40 8 64 12 12 9
AMR [Cq] 22.32–33.56 21.69–36.47 21.20–33.56 24.61–35.47 17.60–35.60 25.26–35.62
Efficiency [%] 109 113 115 111 109 117
Linearity 1.000 1.000 0.996 0.999 0.999 0.997
IVT-RNA LoD [Cq] 34.64 35.21 35.44 34.91 34.97 36.51
LoD [molecules] 18 18 18 18 18 5
AMR [Cq] 17.34–34.64 18.52–35.21 17.56–35.44 19.27–34.91 17.9–34.97 18.87–36.51
Efficiency [%] 106 109 101 103 107 103
Linearity 1.000 0.999 1.000 0.999 0.999 1.000