用于快速检测血管紧张素原基因多态性的实时聚合酶链反应
Real time PCR for fast detection of the angiotensinogen polymorphisms.
作者信息
Dandana Azza, Gammoudi Imen, Ferchichi Salima, Chahed Hinda, Bricca Giampiero, Limem Halima Ben, Maatouk Faouzi, Miled Abdelhedi
机构信息
Department of clinical biochemistry, CHU-Farhat Hached, Sousse, Tunisia.
出版信息
Ann Biol Clin (Paris). 2011 Jul-Aug;69(4):447-51. doi: 10.1684/abc.2011.0598.
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.
目的
开发一种快速可靠的实时聚合酶链反应（PCR）以检测血管紧张素原（AGT）的多态性，比较诱变分离PCR（MS-PCR）和实时PCR这两种方法来确定血管紧张素原基因的M235T、A（-6）G和A（-20）C三种多态性。
方法
在罗氏LightCycler1型定量PCR仪上使用SYBR Green I开发实时PCR方法。我们使用了两条正义引物和一条无义引物。通过比较熔解曲线来检测血管紧张素原基因的多态性。
结果
采用实时PCR和MS-PCR两种方法对DNA样本进行分析。在我们的研究中，两种技术之间未发现差异。
讨论
实时PCR是检测AGT基因M235T、A（-6）G和A（-20）C多态性的一种快速可靠的方法。
结论
这种实时PCR方法是一种可靠的基因检测方法，快速且成本低廉，可在实际中用于研究与心血管疾病相关的AGT基因特定多态性。
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