A review on challenges in the development of multiplex assay for quantitative estimation of viral pathogens

HBV, HCV, and HIV are the primary infectious agents that can be transmitted via exposure to body fluids and affect human health to various degrees. Controlling the spread of these viruses to uninfected individuals is a major task. These viruses have highly competent replication potential and are undetectable by serological markers at an early phase of infection. Molecular techniques on the other hand, by their enhanced sensitivity, selectivity, short detection time, absolute quantification with unparalleled precision have revolutionized the area of diagnosing multiple viral infections simultaneously. Furthermore, detecting multiple viral infections in a single assay is time saving and cost effective as compared to assays designed to detect an individual pathogen. Various digital and real-time PCR-based qualitative and quantitative assays have been developed for estimation of these viruses simultaneously and individually, and very few of them are licensed by the food and drug administration (FDA). Furthermore, the alleged potential of nanotechnology and digital droplet technology has attracted the attention of viral diagnostic research across the globe. In this review, we describe various commercially available technologies for identification and quantification of these viruses and the challenges involved in the development of a cost-effective multiplex assay for simultaneous detection of these viral pathogens.