Selective Detection of Dengue Virus Serotypes Using Tandem Toehold-Mediated Displacement Reactions

Dengue virus (DENV) is the most common human arboviral infection worldwide and can present with severe clinical manifestations. Timely DENV detection improves clinical outcomes, and identification of the DENV serotype (DENV-1-4) may provide beneficial epidemiologic data to inform the initiation of control measures. Here, DENV RNA-triggered, enzyme-free tandem toehold-mediated displacement reactions were developed to identify and serotype DENV in RNA controls and contrived samples through the amplification of a fluorescent signal detected by the use of a fluorescent scanner and a confocal microscope. Each DENV serotype was detected selectively using both imaging methods. In addition, a 384-well plate was used to prepare an array for diagnosis of the four DENV RNA serotypes from contrived clinical samples. The four serotypes of dengue virus were detected using novel enzyme-free amplification reactions, which are more facile than amplification using reverse transcriptase PCR.