Angewandte Chemie International Edition,
Год журнала:
2024,
Номер
unknown
Опубликована: Окт. 12, 2024
The
CRISPR/Cas
system
is
a
powerful
genome
editing
tool
and
possesses
widespread
applications
in
molecular
diagnostics,
therapeutics
genetic
engineering.
But
easy
folding
of
the
target
sequences
causes
remarkable
deterioration
recognition
shear
efficiency
case
single
Cas-CRISPR
RNA
(crRNA)
duplex.
Here,
we
develop
cooperative
shearing
(CRISPR-CS)
system.
Compared
with
traditional
system,
two
CRISPR/Cas-crRNA
duplexes
simultaneously
recognize
different
sites
sequence,
increasing
possibility
efficiency.
Cooperative
cuts
more
methylene
blue-ssDNA
reporters
on
electrode,
enabling
unamplified
nucleic
acid
electrochemical
assay
less
than
5
minutes
detection
limit
9.5×10
Analytical Chemistry,
Год журнала:
2024,
Номер
96(6), С. 2676 - 2683
Опубликована: Янв. 30, 2024
Sepsis
is
an
extremely
dangerous
medical
condition
that
emanates
from
the
body's
response
to
a
pre-existing
infection.
Early
detection
of
sepsis-inducing
bacterial
infections
can
greatly
enhance
treatment
process
and
potentially
prevent
onset
sepsis.
However,
current
point-of-care
(POC)
sensors
are
often
complex
costly
or
lack
ideal
sensitivity
for
effective
detection.
Therefore,
it
crucial
develop
rapid
sensitive
biosensors
on-site
bacteria.
Herein,
we
developed
graphene
oxide
CRISPR-Cas12a
(GO-CRISPR)
biosensor
bacteria
in
human
serum.
In
this
strategy,
single-stranded
(ssDNA)
FAM
probes
were
quenched
with
single-layer
(GO).
Target-activated
Cas12a
trans-cleavage
was
utilized
degradation
ssDNA
probes,
detaching
short
GO
recovering
fluorescent
signals.
Under
optimal
conditions,
employed
our
GO-CRISPR
system
Salmonella
Typhimurium
(S.
Typhimurium)
as
low
3
×
103
CFU/mL
serum,
well
good
specificity
toward
other
competing
addition,
exhibited
excellent
S.
spiked
The
offers
superior
rapidity
has
potential
early
resource-limited
settings,
expediting
patients
at
risk
Analytical Chemistry,
Год журнала:
2022,
Номер
94(17), С. 6566 - 6573
Опубликована: Апрель 22, 2022
Direct,
rapid,
sensitive,
and
selective
detection
of
nucleic
acids
in
complex
biological
fluids
is
crucial
for
medical
early
diagnosis.
We
herein
combine
the
trans-cleavage
ability
clustered
regularly
interspaced
short
palindromic
repeats
(CRISPR)/Cas12a
with
Au-nanobeacon
to
establish
a
CRISPR-based
biosensor,
providing
rapid
miRNA
high
speed
attomolar
sensitivity.
In
this
strategy,
we
first
report
that
activity
CRISPR/cas12a,
which
was
previously
reported
be
triggered
only
by
target
ssDNA
or
dsDNA,
can
activated
directly.
Therefore,
method
direct,
i.e.,
does
not
need
conversion
into
its
complementary
DNA
(cDNA).
Meanwhile,
as
compared
traditional
reporters
molecular
beacon
(MB)
reporters,
exhibit
improved
reaction
kinetics
assay,
miRNA-21
could
detected
very
sensitivity
5
min.
Finally,
proposed
strategy
enables
determination
samples,
potential
tool
Journal of Medical Virology,
Год журнала:
2025,
Номер
97(2)
Опубликована: Фев. 1, 2025
ABSTRACT
Globally
≤
4
billion
of
the
population
are
at
potential
risk
contracting
dengue
virus
(DENV)
infection.
Seasonal
outbreaks
frequently
reported
causing
a
high
healthcare
burden.
Undiagnosed
DENV
can
lead
to
severe
morbidity
and
mortality.
Early
diagnosis
relies
on
molecular
methods,
which
impractical
in
resource‐constrained
settings
(RCSs).
Dengue
be
caused
by
any
four
distinct
serotypes.
Therefore,
simple
method
for
rapid
Pan‐DENV
serotypes
is
utmost
importance
RCSs.
A
fluorescence
detection
platform
using
RT‐RPA
CRISPR/Cas12a
was
developed
targeting
nonstructural
1
(
NS1
)
gene
DENV‐1,
2,
3,
envelope
E
DENV‐2.
Further,
crRNA
specific
were
designed
facilitate
detection.
Analytical
sensitivity
determined
synthetic
RNA
genome.
Clinical
validation
assay
performed
extracted
from
AES/AFI
clinical
samples.
The
CRISPR/Cas12a‐based
detect
all
viz
1−4
single
pot
This
showed
limit
≥
781
zg
reaction
−
,
1.81
ag
−1
62.5
fg
2.5
pg
DENV‐2,
DENV‐3,
DENV‐4
template,
respectively.
Our
demonstrated
analytic
10
ng
DENV‐1
DENV‐4,
0.5
DENV‐3
genomes.
no
cross‐reactivity
with
other
related
etiologies
tested
AFI/AES.
With
76
samples
(DENV
PCR
positive
=
16,
negative
60),
93.7%
100%
specificity
an
overall
accuracy
98.7%
displayed
comparable
results
that
RT‐PCR.
ease
interpretation
Pan‐DENV,
represents
as
ideal
point‐of‐care
test.
upon
field‐deployment
could
help
reducing
burden,
provide
differential
support
initiating
early
prompt
treatment
patients
RCS.
Analytical Chemistry,
Год журнала:
2022,
Номер
94(23), С. 8506 - 8513
Опубликована: Май 30, 2022
CRISPR-based
biosensors
have
attracted
increasing
attention
in
accurate
and
sensitive
nucleic
acid
detection.
In
this
work,
we
report
a
CRISPR/Cas12a-triggered
chemiluminescence
enhancement
biosensor
for
the
ultrasensitive
detection
of
acids
by
introducing
tyramide
signal
amplification
first
time
(termed
CRICED).
The
hybrid
chain
DNA
(crDNA)
formed
NH2-capture
(capDNA)
biotin-recognition
(recDNA)
was
preferentially
attached
to
magnetic
beads
(MBs),
streptavidin–HRP
subsequently
introduced
obtain
MB@HRP-crDNA.
presence
target,
activated
CRISPR/Cas12a
is
capable
randomly
cutting
initiator
(intDNA)
into
vast
short
products,
thus
fractured
intDNA
could
not
trigger
toehold-mediated
DNA-strand
displacement
reaction
(TSDR)
event
with
After
addition
tyramine–AP
H2O2,
abundant
HRP–tyramine–AP
emerges
through
covalent
attachment
HRP–tyramine,
exhibiting
enhanced
(CL)
signals
or
visual
image
readouts.
By
virtue
biosensor,
achieved
high
sensitivity
synthetic
target
amplified
plasmid
using
recombinase
polymerase
(RPA)
as
low
17
pM
single-copy
detection,
respectively.
Our
proposed
CRICED
further
evaluated
test
20
HPV
clinical
samples,
showing
superior
87.50%
specificity
100.00%.
Consequently,
platform
be
an
attractive
means
imaging
holds
promising
strategy
practical
application
diagnostics.
