Microsatellite-based detection of bladder cancer by analysis of urine using fast capilary array electrophoresis and fluorescent primers

Recently our group described the feasibility of polymerase chain reaction (PCR)-based microsatellite analysis of DNA from urine sediment. Bladder cancer and recurrences could be detected with a specificity of 100% and a sensitivity of 90%. Because standard slab gel electrophoresis of PCR products is extremely time consuming we took thought how to develop sensitive, rapid and accurate methods and apparatus to introduce our test rapidly into the clinical setting. Here we demonstrate microsatellite-based detection of bladder cancer using fast capillary array electrophoresis. DNA from blood, urine and bladder tumors was analyzed using PCR and a panel of 20 γ-[³²P] ATP end-labeled or two-color energy transfer fluorescent labeled primers. Slab gel electrophoresis or capillary assay electrophoresis was performed and the results were compared. After a rapid separation (≤ 35 minutes) on capillary arrays the allelic ratios could be quantitatively determined with a precision of ±10%. The estimated precision of standard slab gel electrophoresis is about ±30% after a separation time of 3 to 4 hours. The results of manual performed slab gel analysis could be reliably reproduced by the capillary array. This method provides a significant improvement in the speed, ease and precision of microsatellite-based analyses of bladder cancer. It is more sensitive and already automated which promises the possibility of a wide use in clinical diagnostic and follow up of bladder cancer patients.