Detection of PAX5 mRNA expression in haematological tumor cell lines by real-time reverse transcription polymerase chain reaction

Objective To develop a real-time reverse transcription polymerase chain reaction(RT-PCR) for the relative quantitation of PAX5 and CD19 mRNA expression in 6 haematological tumor cell lines. Methods A serial dilution cDNA reversely transcriped from total RNA in NAMALWA were used to construct standard curves for the PAX5, CD19 and GAPDH amplifications. The amplification efficiencies were identical for both the target gene(PAX5 and CD19) and house keeping gene(GAPDH). Results PAX5 and CD19 mRNA expression level was 2.35% and 2.52% respectively in NAMALWA(B-cell lines), but almost not detectable in other T-and myeloid cell lines. Conclusions The real-time RT-PCR is simple, rapid and convenient for quantification of PAX5 and CD19 mRNA levels, and is suitable for investigation in larger groups of patients with different haematological malignancies.