A MICROPLATE ASSAY FOR K-RAS GENOTYPING

The paper presents a microplate hybridisation assay for the detection of codon 12 mutations of the K-ras protooncogene. Single-stranded targ et DNA, obtained from amplifying sample DNA with 5'-biotin and 5'-digo xigenin-labelled primers and subsequent strand separation, is hybridis ed with solid phase-fixed capture probes complementary to wild-type an d mutated forms of K-ras. After stringent washing the duplex DNA is de tected by an ELISA-like protocol incorporating photometric, fluorometr ic or luminometric detection. Application examples are shown in which the K-ras status was examined in peripheral blood cells. cell cultures , fresh and paraffin-embedded tumour tissue and in stool samples.