G-PoC sensors indeed would present some unique advantages, such as a simple operation (performable by untrained personnel), rapid results, low power supply and use of reagents, and convenience of use specifically in areas where resources are limited ( Arduini et al., 2016 Chan et al., 2017). Therefore, the development of new molecular methods allowing rapid, sensitive, and simple detection of pathogens would be a significant breakthrough in molecular diagnostics opening new technological frontiers in the genome detection, as represented by the Genetic Point of Care (G-PoC) testing ( Petralia and Conoci, 2017 Wang et al., 2021). In addition, it has become evident that COVID-19 antigen rapid diagnostic tests, although being fast and relatively cheap, yield a high percentage of false negative results. This is proved by the diffused difficulty to perform massive and real time diagnosis of coronavirus disease 2019 (COVID-19) for prompt infection management and prevention which is stimulating new approaches and methodologies for molecular detection ( Cheong et al., 2020 Fan et al., 2021 Nunez-bajo et al., 2020). This represents a strong limitation for its massive use, limiting the potential of genome analysis for human health. Therefore, although current PCR-based methods are well-established and consolidated, they are not suitable to be used by unskilled personnel near the patient at competitive costs. However, molecular methods typically need an amplification step achieved through the well-known Polymerase Chain Reaction (PCR) that is intrinsically quite laborious because it involves a multi-step and expensive process (about $15–80 per sample) thus limiting its utilization to specialized laboratories ( Espy et al., 2006). In this context, DNA identification and quantification through molecular methods provides relevant clinical advantages with respect to the traditional laboratory methods, such as bacterial cultures or antibody detection, being much faster (hours versus days for bacterial cultures), specific (allowing the detection of genotypes), sensitive (few copies of pathogens in a sample) and accurate (able to detect different microorganisms through the specific molecular markers, but also their vitality through the mRNA monitoring). The detection of nucleic acids is nowadays of extreme importance in many medical fields for early and accurate diagnosis, personalized therapy, and preventive screening. The detection concept presented here for HBV detection is very versatile and can be extended to other pathogens, paving the way for future development of rapid molecular test for infectious diseases, both viral and bacterial, in Point-of-Care (PoC) format. Thanks to the combination of surface cooperative hybridization scheme with ECL detection strategy, our novel DNA sensor is able to detect HBV genome – both synthetic and extracted – with the unprecedented limit of detection (LoD) of 0.05 cps μL −1 for extracted sample, that is even lower than the typical LoD of PCR methodologies. In this work, we present a novel molecular sensor for the ultrasensitive PCR-free detection of Hepatitis B Virus (HBV) based on electrochemiluminescence (ECL). In this context, the development of innovative detection methods based on signal-amplification rather than analyte-amplification represents a significant breakthrough compared to existing PCR-based methodologies, allowing the development of new nucleic acid detection technologies suitable to be integrated in portable and low-cost sensor devices while keeping high sensitivities, thus enabling massive diagnostic screening. Detection of nucleic acids is crucial in many medical applications, and in particular for monitoring infectious diseases, as it has become perfectly clear after the pandemic infection of COVID-19.
0 Comments
Leave a Reply. |