Abstract
The analysis of viral nucleic acids (NA), DNA or RNA, is a crucial issue in the diagnosis of infections and the treatment and prevention of related human diseases. Conventional nucleic acid tests (NATs) require multistep approaches starting from the purification of the pathogen genetic material in biological samples to the end of its detection, basically performed by the consolidated polymerase chain reaction (PCR), by the use of specialized instruments and dedicated laboratories. However, since the current NATs are too constraining and time and cost consuming, the research is evolving towards more integrated, decentralized, user-friendly, and low-cost methods. These will allow the implementation of massive diagnoses addressing the growing demand of fast and accurate viral analysis facing such global alerts as the pandemic of coronavirus disease of the recent period. Silicon-based technology and microfluidics, in this sense, brought an important step up, leading to the introduction of the genetic point-of-care (PoC) systems. This review goes through the evolution of the analytical methods for the viral NA diagnosis of infection diseases, highlighting both advantages and drawbacks of the innovative emerging technologies versus the conventional approaches.
Highlights
The Coronavirus pandemic has recently focused the attention of researchers worldwide on infectious diseases, requiring the best screening strategy for safeguarding public health.Infectious diseases are disorders provoked by a wide group of microorganisms including viruses, bacteria, fungi, etc., that under certain conditions become dangerous for human safety and capable of rapid transmission among humans or animals
This makes isothermal methods like Loop-Mediated Isothermal Amplification (LAMP), Recombinase Polymerase Amplification (RPA), and Helicase-Dependent Amplification (HDA) faster, as they do not need to reach a high number of thermal cycles but use continuous amplification, which provides detectable amplicons within a few minutes, which is more sensitive than qPCR, since they involve the synergy of multiple specific primers
10 microchannels with a deposited set of primers allowed an LAMP amplification of specific sequences of some equine viruses, such as equine herpesvirus 1 (EHV1) and equine influenza virus (EIV), and optically detected by fluorescence imaging operated by a customized smartphone, revealing down to 5.5 × 104 copies/mL of viral DNA
Summary
The Coronavirus pandemic has recently focused the attention of researchers worldwide on infectious diseases, requiring the best screening strategy for safeguarding public health. Infectious diseases are disorders provoked by a wide group of microorganisms including viruses, bacteria, fungi, etc., that under certain conditions become dangerous for human safety and capable of rapid transmission among humans or animals. This leads to a series of signs and symptoms that are mostly dependent on the type of microorganism causing the infection and the human district involved. Viruses are submicroscopic microorganisms composed of two core elements: the nucleic acid genome, either double- (ds) or single-stranded (ss) DNA or RNA, and a protein-based shell called “capsid” They exist in different habitats but are obliged intracellular parasites, they need to infect a living organism as a host for their growth and replication [1]. A complete overview of the approaches developed for virus detection and infections diagnosis will be given, reporting the state-of-the-art of both the conventional methods and the advancements brought by the emerging new detection technologies
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