Recombinase polymerase amplification combined with a lateral flow dipstick for rapid and visual detection of adenovirus / 军事医学

ABSTRACT

Objective To develop a rapid, accurate, visual, and portable detection method for adenovirus types B (AdvB) and E ( AdvE).Methods Universal primers were targeted on type-specific conserved regions to allow the simultaneous detection of both human Adv (HAdV) species.A detection method based on the combination of recombinase polymerase amplification ( RPA) and lateral flow dipstick ( LFD) was established the sensitivity and specificity evaluated , and throat swab specimens of 19 patients infected with AdvB and AdvE as well as 10 healthy volunteers were detected with this method.Results The detection limit of the method was 10 copies/μl Adv DNA, which was close to that of qPCR , and there were no cross-reactions with other species of Adv and unrelated virus .The detection could be finished within 15 to 20 min within the temperature range of 25 to 45℃.When applied to clinical samples , this method showed 100% sensitivity and specificity.Conclusion This detection assay is a sensitive , specific, rapid and simple method that eliminates the need for expensive equipment , trained personnel or laboratories .The characteristics of this system render it suitable for use in grass-roots healthcare departments , and the system is especially effective for field testing and on-site testing.