Abstract
In
this
study,
we
present
an
ultrasensitive
and
specific
multiplexed
detection
method
for
SARS‐CoV‐2
influenza
(Flu)
utilizing
CRISPR/Cas13a
technology
combined
with
a
hydrogel‐encapsulated
photonic
crystal
(PhC)
barcode
integrated
hybridization
chain
reaction
(HCR).
The
barcodes,
characterized
by
core‐shell
structures,
are
fabricated
through
partial
replication
of
periodically
ordered
hexagonally
close‐packed
silicon
dioxide
beads.
Consequently,
the
opal
hydrogel
shell
these
barcodes
features
abundant
interconnected
pores
that
provide
substantial
surface
area
probe
immobilization.
Furthermore,
inherent
structural
colors
remain
stable
during
events
due
to
robust
mechanical
strength
cores.
This
integration
HCR
leverages
both
highly
RNA
recognition
capabilities
trans‐cleavage
activity
Cas13a
while
employing
enhance
sensitivity.
Upon
encountering
target
RNA,
cleaves
hairpin
probe,
thereby
initiating
subsequent
amplification
enhanced
Our
demonstrates
high
accuracy
sensitivity
in
SARS‐CoV‐2,
Flu
A
B
limit‐of‐detection
as
low
200
aM.
Importantly,
assay
also
exhibits
acceptable
repeated
clinical
sample
testing.
Thus,
our
platform
represents
promising
strategy
sensitive
virus
clinical.
ACS Nano,
Год журнала:
2021,
Номер
15(8), С. 13475 - 13485
Опубликована: Авг. 9, 2021
Nucleic
acid
biomarkers
have
been
widely
used
to
detect
various
viral-associated
diseases,
including
the
recent
pandemic
COVID-19.
The
CRISPR-Cas-based
trans-activating
phenomenon
has
shown
excellent
potential
for
developing
sensitive
and
selective
detection
of
nucleic
acids.
However,
amplification
steps
are
typically
required
when
monitoring
target
is
needed.
To
overcome
aforementioned
challenges,
we
developed
a
CRISPR-Cas12a-based
amplification-free
biosensor
by
surface-enhanced
Raman
spectroscopy
(SERS)-assisted
ultrasensitive
system.
We
integrated
activated
CRISPR-Cas12a
viral
DNA
with
Raman-sensitive
system
composed
ssDNA-immobilized
probe-functionalized
Au
nanoparticles
(RAuNPs)
on
graphene
oxide
(GO)/triangle
nanoflower
array.
Using
this
CRISPR-based
improved
sensitivity
multiviral
DNAs
such
as
hepatitis
B
virus
(HBV),
human
papillomavirus
16
(HPV-16),
HPV-18
an
extremely
low
limit
vast
range
from
1
aM
100
pM
without
steps.
suggest
that
acids
can
be
applied
precise
early
diagnosis
infections,
cancers,
several
genetic
diseases.
Analytical Chemistry,
Год журнала:
2023,
Номер
95(17), С. 7053 - 7061
Опубликована: Апрель 20, 2023
False
positives
and
negatives
in
bioanalytical
assays
remain
a
persistent
problem.
Herein,
multifunctional
photoelectrochemical
(PEC)
biosensor
based
on
ZnIn2S4
(ZIS)/ZnS
quantum
dots
(QDs)@Au–Ag-reversed
photocurrent
of
Cu-metal–organic
framework
(MOF)
coupled
with
CRISPR/Cas-12a-shearing
was
innovatively
developed
for
assay
dual
targets.
First,
Cu-MOF
as
good
PEC
material
shows
cathodic
photocurrent.
Then,
numerous
ZIS/ZnS
QDs
were
assembled
to
the
Au–Ag
nanoparticles
(NPs)
prepare
stable
highly
amplified
signal
probe,
which
can
just
match
energy
level
Cu-MOFs
realized
polarity-reversed
first
time.
As
empty-core
nanostructure
NPs
has
high
specific
surface
area
low
density,
bimetallic
nanocrystal
much
increase
reaction
rate
improve
redox
efficiency.
When
target
CEA-produced
cDNA
opened
hairpin
DNA
(HP1
DNA)
electrode,
QDs@Au–Ag
probe
conjugated
electrode
via
hybridization,
achieving
significantly
reversed
current
CEA
detection.
Moreover,
binding
kanamycin/aptamer
generated
acDNA
(activator),
activate
trans-cleavage
activity
CRISPR-CAS12a
system
ssDNA,
so
sheared
caused
obvious
decrease
kanamycin
The
newly
displayed
excellent
properties
MOF
combined
unique
CRISPR-Cas12a
achieve
sensitive
detection
targets,
open
new
sensing
platform
rapid
accurate
analysis
multiple
targets
effectively
avoid
false
results
clinical
testing.
Heliyon,
Год журнала:
2024,
Номер
10(4), С. e26179 - e26179
Опубликована: Фев. 1, 2024
CRISPR/Cas
systems
have
become
integral
parts
of
nucleic
acid
detection
apparatus
and
biosensors.
Various
such
as
CRISPR/Cas9,
CRISPR/Cas12,
CRISPR/Cas13,
CRISPR/Cas14
CRISPR/Cas3
utilize
different
mechanisms
to
detect
or
differentiate
biological
activities
nucleotide
sequences.
Usually,
CRISPR/Cas-based
are
combined
with
polymerase
chain
reaction,
loop-mediated
isothermal
amplification,
recombinase
amplification
transcriptional
technologies
for
effective
diagnostics.
Premised
on
these,
many
biosensors
been
developed
acids
viral
bacterial
pathogens
in
clinical
samples,
well
other
applications
life
sciences
including
biosecurity,
food
safety
environmental
assessment.
Additionally,
showed
better
specificity
compared
molecular
diagnostic
methods.
In
this
review,
we
give
an
overview
various
methods
highlight
some
advances
their
development
components.
We
also
discourse
operational
challenges
advantages
disadvantages
systems.
Finally,
important
considerations
offered
the
improvement
testing.
Analytical Chemistry,
Год журнала:
2021,
Номер
93(35), С. 11899 - 11909
Опубликована: Авг. 24, 2021
Timely
diagnosis
is
of
great
benefit
to
improve
the
survival
rate
cancer
patients.
Body
fluid
biomarker
detection
a
critical
kind
noninvasive
method
for
diagnosis.
Nevertheless,
traditional
methods
always
rely
on
large-scale
instrument
and
involve
sophisticated
operation.
Clustered
regularly
interspaced
short
palindromic
repeats/CRISPR-associated
protein
(CRISPR/Cas)-based
in
vitro
can
simplify
procedures
sensitivity
specificity,
holding
promise
as
next-generation
molecular
diagnostic
technology.
In
this
Feature,
we
introduce
working
mechanisms
different
kinds
CRISPR/Cas
systems
biosensing
CRISPR/Cas-mediated
strategies
biomarkers
including
nucleic
acids,
proteins,
extracellular
vesicles.
addition,
perspective
challenges
CRISPR/Cas-based
are
discussed.
