Real time PCR for fast detection of the angiotensinogen polymorphisms.

OBJECTIVE to develop a rapid and reliable real-time PCR to detect polymorphisms of angiotensinogen (AGT), to compare the two methods of MS-PCR (Mutagenically Separated PCR) and real-time PCR to determine three polymorphisms of the angiotensinogen gene M235T, the A (-6) G and A (-20) C. METHODS the method of real-time PCR was developed on the PLC Roche LightCycler1 with SYBR Green I. We used two sense primers and a primer nonsense. Detection of polymorphisms of angiotensinogen gene was performed by comparing the melting curves. RESULTS the DNA samples were analyzed by two methods: real-time PCR and MS-PCR. In our study, no differences were found between the two techniques. DISCUSSION The real-time PCR is a rapid and reliable method for detecting gene polymorphisms on the AGT M235T, the A (-6) G and A (-20) C. CONCLUSION this method of real-time PCR is a reliable genetic test, which is fast and cheap and can be used in practice to study particular polymorphisms of AGT gene associated with cardiovascular disease.