Effective screening of infectious
diseases requires a fast, cheap,
and population-scale testing. Antigen pool testing can increase the
test rate and shorten the screening time, thus being a valuable approach
for epidemic prevention and control. However, the overall percent
agreement (OPA) with polymerase chain reaction (PCR) is one-half to
three-quarters, hampering it from being a comprehensive method, especially
pool testing, beyond the gold-standard PCR. Here, a multiantibodies
transistor assay is developed for sensitive and highly precise antigen
pool testing. The multiantibodies capture SARS-CoV-2 spike S1 proteins
with different configurations, resulting in an antigen-binding affinity
down to 0.34 fM. The limit of detection reaches 3.5 × 10–17 g mL–1SARS-CoV-2 spike S1 protein
in artificial saliva, 4–5 orders of magnitude lower than existing
transistor sensors. The testing of 60 nasopharyngeal swabs exhibits
∼100% OPA with PCR within an average diagnoses time of 38.9
s. Owing to its highly precise feature, a portable integrated platform
is fabricated, which achieves 10-in-1 pooled screening for high testing
throughput. This work solves the long-standing problem of antigen
pool testing, enabling it to be a valuable tool in precise diagnoses
and population-wide screening of COVID-19 or other epidemics in the
future.
Rapid screening of infected individuals
from a large population
is an effective means in epidemiology, especially to contain outbreaks
such as COVID-19. The gold standard assays for COVID-19 diagnostics
are mainly based on the reverse transcription polymerase chain reaction,
which mismatches the requirements for wide-population screening due
to time-consuming nucleic acid extraction and amplification procedures.
Here, we report a direct nucleic acid assay by using a graphene field-effect
transistor (g-FET) with Y-shaped DNA dual probes (Y-dual probes).
The assay relies on Y-dual probes modified on g-FET simultaneously
targeting ORF1ab and N genes of SARS-CoV-2 nucleic acid, enabling
high a recognition ratio and a limit of detection (0.03 copy μL
–1
) 1–2 orders of magnitude lower than existing
nucleic acid assays. The assay realizes the fastest nucleic acid testing
(∼1 min) and achieves direct 5-in-1 pooled testing for the
first time. Owing to its rapid, ultrasensitive, easily operated features
as well as capability in pooled testing, it holds great promise as
a comprehensive tool for population-wide screening of COVID-19 and
other epidemics.
Dithieno[3,2-b:2′,3′-d]pyrrole (DTP) derivatives are one of the most important organic photovoltaic materials due to better π-conjugation across fused thiophene rings.
The existing electrochemical biosensors lack controllable
and intelligent
merit to modulate the sensing process upon external stimulus, leading
to challenges in analyzing a few copies of biomarkers in unamplified
samples. Here, we present a self-actuated molecular-electrochemical
system that consists of a tentacle and a trunk modification on a graphene
microelectrode. The tentacle that contains a probe and an electrochemical
label keeps an upright orientation, which increases recognition efficiency
while decreasing the pseudosignal. Once the nucleic acids are recognized,
the tentacles nearby along with the labels are spontaneously actuated
downward, generating electrochemical responses under square wave voltammetry.
Thus, it detects unamplified SARS-CoV-2 RNAs within 1 min down to
4 copies in 80 μL, 2–6 orders of magnitude lower than
those of other electrochemical assays. Double-blind testing and 10-in-1
pooled testing of nasopharyngeal samples yield high overall agreement
with reverse transcription-polymerase chain reaction results. We fabricate
a portable prototype based on this system, showing great potential
for future applications.
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