Analytical Chemistry,
Год журнала:
2024,
Номер
96(2), С. 866 - 875
Опубликована: Янв. 2, 2024
Despite
extensive
efforts,
point-of-care
testing
(POCT)
of
protein
markers
with
high
sensitivity
and
specificity
at
a
low
cost
remains
challenging.
In
this
work,
we
developed
an
aptamer-CRISPR/Cas12a-regulated
liquid
crystal
sensor
(ALICS),
which
achieved
ultrasensitive
detection
using
smartphone-coupled
portable
device.
Specifically,
DNA
probe
that
contained
aptamer
sequence
for
the
target
activation
Cas12a–crRNA
complex
was
prefixed
on
substrate
released
in
presence
target.
The
then
bound
to
activate
collateral
cleavage
reaction,
producing
bright-to-dark
optical
change
DNA-functionalized
interface.
image
captured
by
smartphone
quantification
concentration.
For
two
model
proteins,
SARS-CoV-2
nucleocapsid
(N
protein)
carcino-embryonic
antigen
(CEA),
ALICS
limits
0.4
20
pg/mL,
respectively,
are
higher
than
typical
test
clinical
CEA
test.
sample
tests,
also
exhibited
superior
performances
compared
those
commercial
ELISA
lateral
flow
kits.
Overall,
represents
cost-effective
platform
POCT,
showing
great
potential
pathogen
disease
monitoring
under
resource-limited
conditions.
Communications Biology,
Год журнала:
2025,
Номер
8(1)
Опубликована: Янв. 30, 2025
Critical
to
the
success
of
CRISPR-based
diagnostic
assays
is
selection
a
target
highly
specific
organism
interest,
process
often
requiring
iterative
cycles
manual
selection,
optimisation,
and
redesign.
Here
we
present
PathoGD,
bioinformatic
pipeline
for
rapid
high-throughput
design
RPA
primers
gRNAs
CRISPR-Cas12a-based
pathogen
detection.
PathoGD
fully
automated,
leverages
publicly
available
sequences
scalable
large
datasets,
allowing
continuous
monitoring
validation
primer/gRNA
sets
ensure
ongoing
assay
relevance.
We
designed
five
clinically
relevant
bacterial
pathogens,
experimentally
validated
subset
designs
detecting
Streptococcus
pyogenes
and/or
Neisseria
gonorrhoeae
in
with
without
pre-amplification.
demonstrated
high
specificity
designed,
minimal
off-target
signal
observed
all
combinations.
anticipate
will
be
an
important
resource
current
emerging
pathogens.
on
GitHub
at
https://github.com/sjlow23/pathogd
.
Proceedings of the National Academy of Sciences,
Год журнала:
2025,
Номер
122(11)
Опубликована: Март 10, 2025
Rapid
and
accurate
detection
of
DNA
from
disease-causing
pathogens
is
essential
for
controlling
the
spread
infections
administering
timely
treatments.
While
traditional
molecular
diagnostics
techniques
like
PCR
are
highly
sensitive,
they
include
nucleic
acid
amplification
many
need
to
be
performed
in
centralized
laboratories,
limiting
their
utility
point-of-care
settings.
Recent
advances
CRISPR-based
(CRISPR-Dx)
have
demonstrated
potential
specific
detection,
but
sensitivity
often
constrained
by
slow
trans-cleavage
activity
Cas
enzymes,
necessitating
preamplification
target
acids.
In
this
study,
we
present
a
CRISPR-Cascade
assay
that
overcomes
these
limitations
integrating
positive
feedback
loop
enables
amplification-free
pathogenic
at
atto-molar
levels
achieves
signal-to-noise
ratio
greater
than
1.3
within
just
10
min.
The
versatility
through
bloodstream
infection
pathogens,
including
Methicillin-Sensitive
Staphylococcus
aureus
(MSSA),
Methicillin-Resistant
(MRSA),
Escherichia
coli
,
Hepatitis
B
Virus
(HBV)
spiked
whole
blood
samples.
Additionally,
introduce
multiplexing
OR-function
logic
gate,
further
enhancing
rapid
clinical
Our
findings
highlight
ability
provide
sensitive
paving
way
advanced
applications
beyond.
ACS Sensors,
Год журнала:
2023,
Номер
8(12), С. 4420 - 4441
Опубликована: Ноя. 18, 2023
CRISPR/Cas
system
is
becoming
an
increasingly
influential
technology
that
has
been
repositioned
in
nucleic
acid
detection.
A
preamplification
step
usually
required
to
improve
the
sensitivity
of
CRISPR/Cas-based
The
striking
biological
features
CRISPR/Cas,
including
programmability,
high
and
sequence
specificity,
single-base
resolution.
More
strikingly,
target-activated
trans-cleavage
could
act
as
a
biocatalytic
signal
transductor
amplifier,
thereby
empowering
it
potentially
perform
detection
without
step.
reports
such
work
are
on
rise,
which
not
only
scientifically
significant
but
also
promising
for
futuristic
end-user
applications.
This
review
started
with
introduction
methods
acids
diagnostics
(CRISPR-Dx).
Next,
we
objectively
discussed
pros
cons
steps
CRISPR-Dx.
We
then
illustrated
highlighted
recently
developed
strategies
CRISPR/Cas-powered
amplification-free
can
be
realized
through
uses
ultralocalized
reactors,
cascade
reactions,
ultrasensitive
systems,
or
others.
Lastly,
challenges
perspectives
were
proposed.
It
expected
this
makes
researchers
better
understand
current
emerging
field,
provides
insight
designing
novel
CRISPR-Dx
win
practicable
use
near
future.
ACS Sensors,
Год журнала:
2023,
Номер
8(10), С. 3623 - 3642
Опубликована: Окт. 11, 2023
Over
the
past
few
decades,
pathogens
have
posed
a
threat
to
human
security,
and
rapid
identification
of
should
be
one
ideal
methods
prevent
major
public
health
security
outbreaks.
Therefore,
there
is
an
urgent
need
for
highly
sensitive
specific
approaches
identify
quantify
pathogens.
Clustered
Regularly
Interspaced
Short
Palindromic
Repeats
CRISPR/Cas
systems
Argonaute
(Ago)
belong
Microbial
Defense
Systems
(MDS).
The
guided,
programmable,
targeted
activation
nucleases
by
both
them
leading
way
new
generation
detection.
We
compare
these
two
in
terms
similarities
differences.
In
addition,
we
discuss
future
challenges
prospects
development
biosensors,
especially
electrochemical
biosensors.
This
review
expected
afford
researchers
entering
this
multidisciplinary
field
useful
guidance
provide
inspiration
more
innovative
biosensors
Biosensors,
Год журнала:
2023,
Номер
13(3), С. 410 - 410
Опубликована: Март 21, 2023
Infectious
diseases
contribute
significantly
to
the
global
disease
burden.
Sensitive
and
accurate
screening
methods
are
some
of
most
effective
means
identifying
sources
infection
controlling
infectivity.
Conventional
detecting
strategies
such
as
quantitative
polymerase
chain
reaction
(qPCR),
DNA
sequencing,
mass
spectrometry
typically
require
bulky
equipment
well-trained
personnel.
Therefore,
a
large
population
using
conventional
during
pandemic
periods
often
requires
additional
manpower,
resources,
time,
which
cannot
be
guaranteed
in
resource-limited
settings.
Recently,
emerging
microfluidic
technologies
have
shown
potential
replace
performing
point-of-care
detection
because
they
automated,
miniaturized,
integrated.
By
exploiting
spatial
separation
sites,
platforms
can
enable
multiplex
infectious
reduce
possibility
misdiagnosis
incomplete
diagnosis
with
similar
symptoms.
This
review
presents
recent
advances
used
for
diseases,
including
immunosensors
nucleic
acid
sensors.
As
representative
platforms,
lateral
flow
immunoassay
(LFIA)
polymer-based
chips,
paper-based
devices,
droplet-based
devices
will
discussed
detail.
In
addition,
current
challenges,
commercialization,
prospects
proposed
promote
application
detection.
Critical Reviews in Food Science and Nutrition,
Год журнала:
2023,
Номер
64(32), С. 12061 - 12082
Опубликована: Сен. 10, 2023
Food
safety
problems
have
become
one
of
the
most
important
public
health
issues
worldwide.
Therefore,
development
rapid,
effective
and
robust
detection
is
great
importance.
Amongst
a
range
methods,
nucleic
acid
isothermal
amplification
(NAIA)
plays
role
in
food
detection.
However,
widespread
application
remains
limited
due
to
few
shortcomings.
CRISPR/Cas
system
has
emerged
as
powerful
tool
detection,
which
could
be
readily
integrated
with
NAIA
improve
sensitivity,
specificity,
adaptability
versatility
dependability.
currently
there
was
lack
comprehensive
summary
regarding
integration
field
In
this
review,
recent
advances
based
on
CRISPR/Cas-integrated
were
comprehensively
reviewed.
To
begin
with,
summarized.
Then,
types
working
principles
introduced.
The
applications
for
mainly
introduced
objectively
discussed.
Lastly,
current
challenges
future
opportunities
proposed.
summary,
technology
expected
an
approach
leading
safer
more
reliable
industry.
