Field-Deployable Detection of Genetically Modified Organisms with an Integrated Method of Loop-Mediated Isothermal Amplification and CRISPR/FnCas12a
Journal of Agricultural and Food Chemistry,
Год журнала:
2025,
Номер
73(9), С. 5625 - 5634
Опубликована: Фев. 18, 2025
The
detection
of
genetically
modified
organisms
(GMOs)
is
crucial
for
regulatory
compliance
and
consumer
safety.
This
study
presents
a
novel
method
combining
loop-mediated
isothermal
amplification
(LAMP)
with
CRISPR/Cas12a
cleavage,
termed
Cas-pfLAMP,
sensitive
specific
GMO
detection.
We
developed
assays
three
GM
events:
maize
DBN9936
MON810
soybean
GTS40-3-2.
By
incorporating
universal
protospacer
adjacent
motif
(PAM)
sequence
into
LAMP
primers,
we
overcame
the
limitations
PAM
site
dependence.
Cas-pfLAMP
demonstrated
high
specificity
sensitivity,
limits
as
low
10-12
copies
per
reaction.
Furthermore,
point-of-care
testing
platform
integrating
rapid
DNA
extraction,
lateral
flow
strips
on-site
achieved
comparable
sensitivity
to
qPCR,
detecting
contents
0.1%
in
simulated
samples
within
40
min.
offers
advantages
independence,
suitability
field
without
specialized
equipment.
approach
represents
promising
new
generation
methods
potential
applications
various
scenarios.
Язык: Английский
Ultrasensitive and visual identification of lethal mushroom Russula subnigricans by LAMP-based CRISPR/Cas12a assay
Microchemical Journal,
Год журнала:
2025,
Номер
unknown, С. 113100 - 113100
Опубликована: Фев. 1, 2025
Язык: Английский
Viral Contamination of Food
Elsevier eBooks,
Год журнала:
2025,
Номер
unknown, С. 79 - 92
Опубликована: Янв. 1, 2025
Язык: Английский
An extraction-free one-step pathogen detection system (Ex-opods) based on LAMP and CRISPR-Cas12b
Microchemical Journal,
Год журнала:
2025,
Номер
unknown, С. 113724 - 113724
Опубликована: Апрель 1, 2025
Язык: Английский
Biosensing platforms for DNA diagnostics based on CRISPR/Cas nucleases: towards the detection of nucleic acids at the level of single molecules in non-laboratory settings
Biomeditsinskaya Khimiya,
Год журнала:
2024,
Номер
70(5), С. 287 - 303
Опубликована: Янв. 1, 2024
The
use
of
CRISPR/Cas
nucleases
for
the
development
DNA
diagnostic
systems
in
out-of-laboratory
conditions
(point-of-need
testing,
PONT)
has
demonstrated
rapid
growth
last
few
years,
starting
with
appearance
2017–2018
first
platforms
known
as
DETECTR
and
SHERLOCK.
are
based
on
a
combination
methods
nucleic
acid
isothermal
amplification
selective
detection
target
amplicons.
This
significantly
improves
sensitivity
specificity
PONT,
making
them
comparable
or
even
superior
to
polymerase
chain
reaction,
considered
“gold
standard”
diagnostics.
review
considers
modern
approaches
coupling
using
Cas9,
Cas12a,
Cas12b,
Cas13a,
Cas14,
Cas3
various
amplification,
an
emphasis
works
which
at
level
single
molecules
(attomolar
subattomolar
concentrations
target)
is
achieved.
properties
used
targeted
diagnostics
features
briefly
context
biosensing
platforms.
Special
attention
paid
most
promising
directions
nuclease.
Язык: Английский