Electrochemical DNA biosensors show great potential for the sensitive and sequence‐specific detection of disease pathogens. To date, although there are a large number of published articles showing various sensing strategies of an electrochemical biosensor, only a few of those studies actually reach the commercialization phase, because addressing one technical issue would usually be at the expense of another. Moreover, there are various adoptable formats in electrochemical biosensors. Current approaches may or may not use enzymes, immobilize DNA probes, label the sensing substrates with an electroactive reporter molecule, or any combination of these. Although much has been paid to individual advantages and disadvantages, there are a very limited number Review articles studying and analyzing the synergistic aspects of various detection strategies when combined with each other. It is the aim of this Review to highlight the hotspots for innovation when these strategies are used in tandem, in order to create more streamlined improvements in making a versatile biosensor platform that can be administered at point of care.
Pooled
testing has been widely adopted recently to facilitate large-scale
community testing during the COVID-19 pandemic. This strategy allows
to collect and screen multiple specimen samples in a single test,
thus immensely saving the assay time and consumable expenses. Nevertheless,
when the outcome of a pooled testing is positive, it necessitates
repetitive retesting steps for each sample which can pose a serious
challenge during a rising infection wave of increasing prevalence.
In this work, we develop a unique barcoded primer-assisted sample-specific
pooled testing strategy (Uni-Pool) where the key genetic sequences
of the viral pathogen in a crude sample are extracted and amplified
with concurrent tagging of sample-specific identifiers. This new process
improves the existing pooled testing by eliminating the need for retesting
and allowing the test results–positive or negative–for
all samples in the pool to be revealed by multiplex melting curve
analysis right after real-time polymerase chain reaction. It significantly
reduces the total assay time for large-scale screening without compromising
the specificity and detection sensitivity caused by the sample dilution
of pooling. Our method was able to successfully differentiate five
samples, positive and negative, in one pool with negligible cross-reactivity
among the positive and negative samples. A pooling of 40 simulated
samples containing severe acute respiratory syndrome coronavirus-2
pseudovirus of different loads (min: 10 copies/μL; max: 103 copies/μL) spiked into artificial saliva was demonstrated
in eight randomized pools. The outcome of five samples in one pool
with a hypothetical infection prevalence of 15% in 40 samples was
successfully tested and validated by a typical Dorman-based pooling.
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