Method of analysing a blood sample of a subject for the presence of a disease marker
The present invention relates to a method of analyzing a blood sample of a subject for the presence of a disease marker, said method comprising the steps of a) extracting nucleic acid from anucleated blood cells in said blood sample to provide an anucleated blood cells-extracted nucleic acid fraction, and b) analyzing said anucleated blood cells-extracted nucleic acid fraction for the presence of a disease marker, wherein said disease marker is a disease-specific mutation in a gene of a cell of said subject, or wherein said disease marker is a disease-specific expression profile of genes of a cell of said subject.
1. A method of typing a sample from a subject, comprising:
(a) providing a blood sample from the subject;
(b) isolating thrombocytes from said blood sample;
(c) extracting ribonucleic acid (RNA) from said isolated thrombocytes to provide a thrombocyte-extracted RNA fraction;
(d) detecting at least one of Kirsten rat sarcoma (KRAS), epidermal growth factor receptor (EGFR) and v-raf murine sarcoma viral oncogene homolog B1 (BRAF) RNA in said thrombocyte-extracted RNA fraction by selective amplification of said RNA using at least one mutation-specific amplification primer; and
(e) determining a nucleotide sequence of said detected at least one of KRAS, EGFR and BRAF RNA, thereby typing the sample from the subject.
2. The method of claim 1 , whereby said detecting in step (d) comprises the selective amplification of said RNA by reverse transcriptase polymerase chain reaction amplification.
3. The method of claim 1 , comprising determining a nucleotide sequence of KRAS, EGFR and BRAF RNA.
4. The method of claim 1 , wherein said at least one mutation-specific amplification primer is an oligodeoxyribonucleotide.
5. The method of claim 1 , wherein a nucleotide sequence of at least one of KRAS, EGFR and BRAF RNA is determined with at least one probe comprising a label moiety.