Analytical Chemistry,
Год журнала:
2024,
Номер
96(39), С. 15657 - 15664
Опубликована: Сен. 19, 2024
The
multimode
assay
based
on
multiple
response
mechanisms
has
received
great
attention
to
effectively
improve
the
accuracy
of
a
sensing
platform.
However,
multifunctional
materials
for
simultaneously
satisfying
multiple-mode
detections
are
still
in
shortage
due
incompatibility
signal
transduction
different
modes.
Here,
taking
human
papillomavirus
16
(HPV-16)
DNA
(TDNA)
as
model
its
important
role
cervical
cancer,
novel
material,
ethyl
violet
(EV)@NH
Analytical Chemistry,
Год журнала:
2024,
Номер
96(25), С. 10443 - 10450
Опубликована: Июнь 12, 2024
Due
to
their
ability
selectively
target
pathogen-specific
nucleic
acids,
CRISPR-Cas
systems
are
increasingly
being
employed
as
diagnostic
tools.
"One-pot"
assays
that
combine
acid
amplification
and
(NAAT–CRISPR-Cas)
in
a
single
step
have
emerged
one
of
the
most
popular
biosensing
formats.
However,
operational
simplicity
comes
at
cost,
with
one-pot
typically
less
sensitive
than
corresponding
two-step
NAAT–CRISPR-Cas
often
failing
detect
targets
low
concentrations.
It
is
thought
these
performance
reductions
result
from
competition
between
two
enzymatic
processes
driving
assay,
namely,
Cas-mediated
cis-cleavage
polymerase-mediated
DNA.
Herein,
we
describe
novel
RPA–Cas12a
assay
circumvents
this
issue
by
leveraging
situ
complexation
target-specific
sgRNA
Cas12a
purposefully
limit
concentration
active
during
early
stages
assay.
Using
clinically
relevant
against
DNA
for
HPV-16,
show
how
format
reduces
cleavage
engenders
significant
improvements
detection
when
compared
traditional
format,
even
patient-derived
samples.
Finally,
gain
further
insight
into
use
experimental
data
formulate
mechanistic
model
describing
Cas
enzyme
amplification.
These
findings
suggest
limiting
rates
proteins
viable
strategy
improving
NAAT-CRISPR-Cas
assays.
Analytical Chemistry,
Год журнала:
2024,
Номер
96(39), С. 15657 - 15664
Опубликована: Сен. 19, 2024
The
multimode
assay
based
on
multiple
response
mechanisms
has
received
great
attention
to
effectively
improve
the
accuracy
of
a
sensing
platform.
However,
multifunctional
materials
for
simultaneously
satisfying
multiple-mode
detections
are
still
in
shortage
due
incompatibility
signal
transduction
different
modes.
Here,
taking
human
papillomavirus
16
(HPV-16)
DNA
(TDNA)
as
model
its
important
role
cervical
cancer,
novel
material,
ethyl
violet
(EV)@NH
