Comparison of standard PCR and the LightCycler technique to determine the thrombophilic mutations: an efficiency and...

For several years it has been possible to routinely detect numerous mutations in the human genome by different methods. The most common technique is a standard PCR, but real time fluorescence PCR is increasingly being used. The purpose of this paper is to compare these two different techniques from the point of view of reliability, time consumption, and cost. More than 600 DNA samples of prevalence studies and from cancer patients were used to determine mutations in the genes of coagulation factor V Leiden, prothrombin, and methylenetetrahydrofolate reductase using standard PCR. A subset of 132 samples from the same pool was also tested by LightCycler PCR for the same coagulation gene mutations. Originally LightCycler techniques were applied for quantitative PCR by real time fluorescence measuring. Adding a melting curve analysis allows mutation detection. The results were perfectly concordant. The cost for the reagents is nearly the same for both methods but the time consumption for standard PCR is much higher than for the LightCycler method, resulting in higher laboratory personnel costs.