Point-of-care molecular diagnostic tests show great promise for providing accurate,timely results in lowinfrastructure healthcare settings and at home.The design space for these tests is limited by a variety of possib...Point-of-care molecular diagnostic tests show great promise for providing accurate,timely results in lowinfrastructure healthcare settings and at home.The design space for these tests is limited by a variety of possible background reactions,which often originate from relatively weak promiscuous activities of the enzymes used for nucleic acid amplification.When this background signal is amplified alongside the signal of the intended biomarker,the dynamic range of the test can be severely compromised.Therefore,a detailed knowledge of potential side reactions arising from enzyme promiscuity can improve rational design of point-of-care molecular diagnostic tests.Towards this end,we report a previously unknown synergistic reaction between T7 RNA polymerase and Bsu DNA polymerase that produces nucleic acid in the presence of single-stranded DNA or RNA.This reaction occurs in the absence of any previously reported substrates for either polymerases and compromises a theoretical microRNA amplification scheme utilizing these polymerases.展开更多
基金This work is supported by a generous gift from Kendall and Susan Warren and the NIH Director's New Innovator Award DP2 HL117748-01 to A.P.L.The authors wish to thank Maxwell DeNies,Lili Tian,and Jennifer Knister for helpful discussion.
文摘Point-of-care molecular diagnostic tests show great promise for providing accurate,timely results in lowinfrastructure healthcare settings and at home.The design space for these tests is limited by a variety of possible background reactions,which often originate from relatively weak promiscuous activities of the enzymes used for nucleic acid amplification.When this background signal is amplified alongside the signal of the intended biomarker,the dynamic range of the test can be severely compromised.Therefore,a detailed knowledge of potential side reactions arising from enzyme promiscuity can improve rational design of point-of-care molecular diagnostic tests.Towards this end,we report a previously unknown synergistic reaction between T7 RNA polymerase and Bsu DNA polymerase that produces nucleic acid in the presence of single-stranded DNA or RNA.This reaction occurs in the absence of any previously reported substrates for either polymerases and compromises a theoretical microRNA amplification scheme utilizing these polymerases.
