Abstract
The
development
of
novel
method
for
drug‐resistant
bacteria
detection
is
imperative.
A
simultaneous
dual‐gene
Test
methicillin‐resistant
Staphylococcus
aureus
(MRSA)
developed
using
an
Argonaute‐centered
portable
biosensor
(STAR).
This
the
first
report
concerning
Argonaute‐based
pathogenic
detection.
Simply,
species‐specific
mec
and
nuc
gene
are
isothermally
amplified
loop‐mediated
isothermal
amplification
(LAMP)
technique,
followed
by
enabled
its
programmable,
guided,
sequence‐specific
recognition
cleavage.
With
strategy,
targeted
nucleic
acid
signals
dexterously
converted
into
fluorescent
signals.
STAR
capable
detecting
mecA
simultaneously
in
a
single
reaction.
limit
10
CFU/mL
with
dynamic
range
from
to
7
CFU/mL.
sample‐to‐result
time
<65
min.
successfully
adapted
detect
clinical
samples,
contaminated
foods,
MRSA‐infected
animals.
work
broadens
reach
biosensing
presents
bacterial
point‐of‐need
(PON)
platform.
ACS Sensors,
Год журнала:
2023,
Номер
8(3), С. 1076 - 1084
Опубликована: Янв. 18, 2023
Next-generation
biosensing
tools
based
on
CRISPR/Cas
have
revolutionized
the
molecular
detection.
A
number
of
CRISPR/Cas-based
biosensors
been
reported
for
detection
nucleic
acid
targets.
The
establishment
efficient
methods
non-nucleic
target
would
further
broaden
scope
this
technique,
but
up
to
now,
concerning
research
is
limited.
In
current
study,
we
a
versatile
platform
small-molecule
called
SMART-Cas12a
(small-molecule
aptamer
regulated
test
using
CRISPR/Cas12a).
Simply,
hybridization
chain
reaction
cascade
signal
amplification
was
first
trigged
by
functional
(aptamer)
through
binding.
Then,
system
integrated
recognize
amplified
products
followed
activation
trans-cleavage.
As
such,
can
be
ingeniously
converted
signals
and
then
fluorescent
that
readily
visualized
analyzed
customized
3D-printed
visualizer
with
help
home-made
App-enabled
smartphone.
Adenosine
triphosphate
selected
as
model
target,
under
optimized
conditions,
achieved
fine
analytical
performance
linear
range
from
0.1
750
μM
limit
1.0
nM.
satisfactory
selectivity
recoveries
obtained
demonstrated
method
suitable
complex
sample
environment.
sample-to-answer
time
less
than
100
min.
Our
work
not
only
expanded
reach
CRISPR-Cas
in
also
provided
prototype
generalized
detecting
wider
analytes
desirable
adaptability,
sensitivity,
specificity,
on-site
capability.
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.
Environmental Technology & Innovation,
Год журнала:
2024,
Номер
34, С. 103625 - 103625
Опубликована: Апрель 4, 2024
Contaminants,
such
as
nucleic
acids
or
toxic
small
molecules,
threaten
both
human
health
and
ecosystems
when
they
infiltrate
the
environment.
The
precise
highly
sensitive
identification
of
contaminants
holds
paramount
importance
across
diverse
domains,
including
safeguarding
food
integrity,
facilitating
clinical
diagnostics,
monitoring
environmental
conditions.
Traditional
methodologies,
encompassing
spectroscopy,
chromatography,
sequencing,
metagenomics,
have
conventionally
served
pivotal
roles
in
detection
processes.
Nevertheless,
these
methods
encountered
recurring
challenges
related
to
sensitivity,
specificity,
portability.
This
review
focuses
on
groundbreaking
CRISPR/Cas12-based
biosensors.
These
biosensors
leverage
incredible
precision
programmability
CRISPR/Cas
system
recognize
specific
targets.
Here,
we
comprehensively
assess
fundamental
mechanisms
that
enable
detection,
ranging
from
guide
RNA
design
collateral
cleavage.
versatility
CRISPR/Cas12
becomes
evident
through
their
applications.
applications
encompass
medical
safety,
monitoring.
transition
conventional
ultimately
represents
a
significant
milestone
contaminant
detection.
By
incorporating
molecular
biology,
nanotechnology,
bioinformatics,
potential
reshape
landscape
water
CRIPSR-Cas
diagnostics
is
transformative
technology
paves
way
for
safer
healthier
future
environment
life.
Heliyon,
Год журнала:
2025,
Номер
11(2), С. e42013 - e42013
Опубликована: Янв. 1, 2025
The
issue
of
antimicrobial
resistance
(AMR)
poses
a
major
challenge
to
global
health,
evidenced
by
alarming
mortality
predictions
and
the
diminishing
efficiency
conventional
drugs.
CRISPR-Cas
system
has
proven
be
powerful
tool
in
addressing
this
challenge.
It
originated
from
bacterial
adaptive
immune
mechanisms
gained
significant
recognition
scientific
community.
This
review
aims
explore
applications
technologies
combating
AMR,
evaluating
their
effectiveness,
challenges,
potential
for
integration
into
current
strategies.
We
conducted
comprehensive
recent
literature
databases
such
as
PubMed
Web
Science,
focusing
on
studies
that
employ
against
AMR.
offer
transformative
approach
combat
with
extend
beyond
traditional
Integrating
these
existing
methods
could
significantly
enhance
our
ability
manage
potentially
reverse
growing
problem
resistance.
Future
research
should
address
technical
ethical
barriers
facilitate
safe
effective
clinical
environmental
applications.
underscores
necessity
interdisciplinary
collaboration
international
cooperation
harness
full
fight
superbugs.
Frontiers in Microbiology,
Год журнала:
2023,
Номер
14
Опубликована: Ноя. 23, 2023
The
World
Health
Organization
(WHO)
estimated
that
pathogens
like
Escherichia
coli,
primarily
linked
to
food
and
water
contamination,
are
associated
with
485,000
deaths
from
diarrheal
diseases
annually,
translating
a
staggering
worldwide
economic
loss
of
nearly
12
billion
USD
per
annum.
International
organizations
the
WHO
United
Nations
Children’s
Fund
(UNICEF)
have
established
related
guidelines
criteria
for
pathogenic
detection
technologies
driving
search
innovative
efficient
methods.
This
comprehensive
review
examines
trajectory
waterborne
bacteria
traditional
techniques,
i.e.,
culture-based
methods,
current
methods
including
various
forms
polymerase
chain
reaction
(PCR)
techniques
[qualitative
real-time
PCR,
digital
ELISA,
loop-mediated
isothermal
amplification,
next-generation
sequencing
(NGS)]
emerging
biosensors
artificial
intelligence
(AI).
scope
paper
focuses
on
recognized
as
human
pathogens,
posing
tangible
threats
public
health
through
waterborne.
techniques’
merits,
constraints,
research
gaps
future
perspectives
critically
discussed.
Advancements
in
droplet
NGS
significantly
improved
sensitivity
specificity,
revolutionizing
pathogen
detection.
Additionally,
integration
(AI)
these
has
enhanced
accuracy,
enabling
analysis
large
datasets.
Molecular-based
show
promise
quality
monitoring,
especially
resource-constrained
settings,
but
on-site
practical
implementation
remains
challenge.
pairwise
comparison
metrics
used
this
also
offer
valuable
insights
into
quick
evaluation
advantages,
limitations
focusing
their
applicability
field
settings
timely
analyses.
Future
efforts
should
focus
developing
robust,
cost-effective
user-friendly
routine
ultimately
safeguarding
global
supplies
health,
AI
data
playing
crucial
role
advancing
safer
environment.
