Advanced multiplexed electrochemical nanobiosensors for simultaneous detection of BK polyomavirus miRNAs in renal transplants

Abstract

BK Polyomavirus (BKPyV)-associated nephropathy (BKPyVAN) is a leading cause of kidney transplant graft loss. Although polymerase chain reaction (PCR) is a common technique for quantifying BK Polyomavirus (BKPyV) viral loads, it lacks optimal sensitivity due to variations in sample type and source, DNA extraction techniques, primer and probe sequences, and the BKPyV strain DNA used for standard-curve genotype variance, all of which can affect the results. Studies have shown that bkv-miR-B1-5p and bkv-miR-B1-3p can be suitable candidates for the diagnosis of nephropathy in kidney transplant recipients, especially in the early stages of the disease. This research introduces an innovative electrochemical nanobiosensor that can detect BKPyV-related miRNAs, including bkv-miR-B1-5p and bkv-miR-B1-3p, with high sensitivity in renal transplant patients. By using a biotin-modified molecular beacon (biotin-MB) and gold nanoparticles (AuNP)/silver nanoparticles (AgNP), this sensor is able to detect viral miRNA with unparalleled sensitivity. Using square-wave voltammetry (SWV), the system can detect miRNA targets simultaneously, surpassing the constraints of traditional techniques like PCR. The sensor displays high linearity with detection limits of 0.3 fM for bkv-miR-B1-5p and 3.6 fM for bkv-miR-B1-3p. This technology offers the potential for diagnosing BKPyVAN earlier and more accurately, offering important information to enhance the lifespan of kidney transplants.