Microbial Biotechnology,
Journal Year:
2025,
Volume and Issue:
18(4)
Published: April 1, 2025
ABSTRACT
We
developed
a
sensitive
and
specific
method
based
on
recombinase‐aided
amplification
(RAA)
clustered
regularly
interspaced
short
palindromic
repeats
(CRISPR)‐CRISPR‐associated
protein
13a
(Cas13a).
This
method,
named
CRISPR‐based
Rapid
Efficient
Test
(CRISPRET),
is
designed
for
the
early
diagnosis
of
Influenza
B
(FluB)
with
aim
shortening
its
transmission
chain.
identified
conserved
regions
in
Virus
(IBV)
NS
gene
forward
reverse
primers
along
crRNAs.
then
established
optimised
reaction
system,
Nucleic
Acid
Positive
Reference
Materials
IBV
were
used
to
evaluate
detection
limit
(DL)
CRISPRET.
Additionally,
we
collected
257
clinical
samples,
comprising
127
samples
from
patients
infection
130
healthy
individuals,
subjected
them
dual
using
CRISPRET
qPCR
positive
predictive
value
(PPV),
negative
(NPV),
sensitivity
specificity
one
primer,
two
primers,
crRNAs
establish
optimise
CRISPR
ET.
The
demonstrated
DL
500
copies·μL
−1
when
assisted
by
appropriate
equipment.
Despite
requiring
auxiliary
equipment
30‐min
reaction,
ET
enables
nucleic
acid
within
approximately
first
5
min,
achieving
high
(100%),
(97.69%),
PPV
(97.69%)
NPV
concordance
rate
98.83%
qPCR.
offers
simple,
field‐applicable,
one‐step
rapid
IBV.
It
has
strong
potential
field‐testing
applications
intelligent
integration
into
existing
diagnostic
systems.
