Clinical application of real-time fluorescence quantitative PCR for the detection of fecal tumor M2-pyruvate kinase in colorectal cancer patients / 国际检验医学杂志

ABSTRACT

Objective To investigate the application valve of real-time fluorescence quantitative polymerase chain reaction(RT-PCR) for the detection of tumor M2-pyruvate kinase(tM2-PK) DNA in patients with colorectal cancer(CRC).Methods Fragment of tM2-PK DNA(162 bp) was amplified and inserted into PGM-T vector to construct recombinant plasmid,which was used to develop RT-PCR method.Sensitivity,specificity and repeatability of RT-PCR for the detection of tM2-PK were analyzed.From Jan.2014 to Jun.2016,200 CRC patients and 100 healthy subjects were enrolled and detected for fecal and serum tM2-PK DNA by using RT-PCR,and the detected results were compared with those detected by using enzyme linked immunosorbent assay(ELISA).Results Recombinant plasmid was successfully constructed,which was certified by sequencing.The sensitivity of RT-PCR for the detection of tM2-PK DNA was 10 copy/mL,with high specificity and 0.3%-2.9% of coefficient of variation.In patients,the positive rate of fecal tM2-PK DNA,detected by RT-PCR,was 92.50%,and that of ELISA to detect tM2-PK was 80.00%.Fecal and serum levels of tM2-PK were correlated with the pathologic stages of tumour.Conclusion Self-established RT-PCR could be specificity and sensitivity for the detection of fecal tM2-PK,which could be used for the early diagnosis of CRC.