A Mismatch-Tolerant Reverse Transcription Loop-Mediated Isothermal Amplification Method and Its Application on Simultaneous Detection of All Four Serotype of Dengue Viruses.

ABSTRACT: Loop-mediated isothermal amplification (LAMP) has been widely used in the detection of pathogens causing infectious diseases. However, mismatches between primers (especially in the 3'-end) and templates significantly reduced the amplification efficiency of LAMP, and limited its application to genetically diverse viruses. Here, we reported a novel mismatch-tolerant LAMP assay and its application in the detection of dengue viruses (DENV). The novel method features the addition of as little as 0.15 U of high-fidelity DNA polymerase to the standard 25 μl LAMP reaction mixture. This amount was sufficient to remove the mismatched bases at the 3'-end of primers, thereby resulting in excellent tolerance for various mismatches occurring at the 3'-end of the LAMP primers during amplification. This n

SUBMITTER: Zhou Y 

PROVIDER: S-EPMC6518337 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature