核酸
化学
克莱诺碎片
聚合酶
核酸内切酶
DNA聚合酶
底漆(化妆品)
DNA
环介导等温扩增
分子信标
分子生物学
生物化学
重组酶聚合酶扩增
寡核苷酸
生物
核酸外切酶
有机化学
作者
Pintao Li,Qiuyuan Lin,Huiwen Xiong,Jilie Kong,Xin Ye,Xueen Fang
标识
DOI:10.1021/acs.analchem.3c03261
摘要
In this study, we developed an innovative highly specific nucleic acid isothermal detection assay based on prokaryotic DNA polymerase I with exquisitely designed fluorescent probes, achieving high sensitivity and 100% specificity within 30 min. The fluorescent nucleic acid probe was designed and constructed based on the specific flap cleavage endonuclease activity of prokaryotic DNA polymerase I (including the Bst, Bsu, Bsm, and Klenow DNA polymerases). The flap endonuclease activity depends on the length of the flap DNA and polymerization activity, which greatly reduces the false-positive rate caused by primer dimerization. This robust assay was also validated by the detection of rotavirus with great specificity and sensitivity. It could be a great alternative to qPCR in the field of point-of-care detection of pathogens.
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