Shortening distance of forward and reverse primers for nucleic acid isothermal amplification
文献类型： 外文期刊
作者： Qu Haitao 1 ; Zhang Wenchao 1 ; Zhang Xiaohui 1 ; Wang Xiujun 2 ; Li Sulong 3 ;
作者机构： 1.Harbin Dege Biotechnol Co Ltd, Harbin 150000, Peoples R China
2.Heilongjiang Acad Agr Sci, Harbin 150086, Peoples R China
3.Heilongjiang Entry Exit Inspect & Quarantine Bur, Harbin 150001, Peoples R China
关键词： DNA polymerase;DNA replication;DNA structure;gene amplification.
期刊名称：BIOLOGICAL CHEMISTRY （ 影响因子：3.915； 五年影响因子：3.922 ）
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收录情况： SCI
摘要： Existent nucleic acid isothermal detection techniques for clinical diseases are difficult to promote greatly due to limitations in such aspects as methodology, costs of detection, amplification efficiency and conditions for operation. There is therefore an urgent need for a new isothermal amplification method with the characteristics of high accuracy, easy operation, short time of detection and low costs. We have devised a new method of nucleic acid isothermal amplification using Bst DNA polymerase under isothermal conditions (60-65°C). We call this method of amplification by shortening the distance between forward and reverse primers for nucleic acid isothermal amplification SDAMR The results demonstrated that this technique is highly sensitive, specific and has short reaction times (40-60 min). Results of sequencing show that the products of SDAMP amplification are mainly polymers formed by series connection of monomers formed through linkage of forward primer and complementary sequences in reverse primer via a few bases. The method is different from current methods of nucleic acid amplification. Our study shows, however, that it is a specific method of nucleic acid isothermal amplification depending on interactions between primers and DNA template.
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