A RT-LAMP based hydrogen ion selective electrode sensing for effective detection HIV-1 RNA with high-sensitivity

Abstract

Development of a simple, sensitive and specific method for HIV-1 assay is demanded. In some emergency situations, a portable reverse transcription loop-mediated isothermal amplification (RT-LAMP) platform is necessary. A novel hydrogen ion-selective RT-LAMP (H+-RT-LAMP) sensor was developed for highly sensitive HIV-1 using hydrogen ion concentration as a signal producing compound. The H+-RT-LAMP sensor was based on screen-printed with carbon-doped graphene conductive layer, carbon nanotubes (CNTs) solid electrolyte layer, and reference electrode KCl functional layer, which provided superior electrical signals, high sensitivity and stability for HIV-1 detection. The rate of hydrogen ion production was related to the viral load (VL) during the amplification of HIV-1. The sensitive HIV-1 RNA detection range was achieved from 0 copies/μL to 105 copies/μL. The proposed H+-RT-LAMP sensor was applied to detect HIV-1 RNA in real human serum. 57 samples were detected and satisfactory results were obtained. Compared with commercial instruments, the detection results of H+-RT-LAMP sensor demonstrated a perfect concordance rate of 96.5 %, and the limit of detection (LOD) for the clinical samples was 10 copies per tube. Time to get the result is from 10 min to 25 min. Therefore, the proposed H+-RT-LAMP sensor was particularly suitable as a point-of-care device for the on-site detection of HIV-1.