Detection of disseminated tumor cells by quantitative RT-PCR of cytokeratin 20 mRNA

摘要

A method for the detection of cytokeratin 20 (CK-20) mRNA in the tissue or body fluid sample of a patient, which is characterized by i) isolating RNA from said sample; ii) preparing cDNA from said RNA by reverse transcription and dividing said cDNA into two compartiments; iii) performing polymerase chain reaction (PCR) with CK-20 specific primers in the first compartiment; iv) performing PCR with primers specific for a housekeeping gene in the second compartiment; v) calculating the relative ratio of mRNA of CK-20 to the housekeeping gene from the results of steps iii) and iv) as means for amount of the CK-20 mRNA in the sample, provides a sensitive and quantitative method for the detection of CK-20.