Establishment and Application of a Digital PCR Method for Coxsackievirus A6
Abstract
Background
The
emergence
of
Coxsackievirus
A6
recombinant
strains
may
result
in
a
significant
public
health
burden
due
to
hand,
foot,
and
mouth
disease.
Traditional
quantitative
Polymerase
Chain
Reaction
(qPCR)
methods
are
widely
used
for
laboratory
diagnosis
but
have
low
sensitivity
cannot
provide
accurate
quantification.
This
study
established
evaluated
droplet
digital
(dPCR)
method
the
detection
A6,
aiming
achieve
precise
pathogen
detection.
Methods
Specific
primers
probes
were
synthesized,
plasmid
standard
was
used.
dPCR
optimized
by
adjusting
primer
concentration,
probe
cycle
number.
evaluation
specificity,
sensitivity,
reproducibility,
47
clinical
sample
testing
conducted,
performance
compared
with
results
real-time
fluorescent
Reaction.
Results
A
successfully
established,
lower
limit
ranging
from
5.23
10.55
copies/µL.
There
no
cross-reactivity
other
enteroviruses
besides
A6.
Repeated
measurements
samples
above
minimum
concentration
showed
relative
deviation
value
less
than
25%,
demonstrating
excellent
specificity
reproducibility.
Testing
using
method,
qPCR
as
gold
standard,
that
true
positive
rate
92.86%,
negative
100%,
Kappa
coefficient
0.91.
Conclusions
developed
an
absolute
quantification
nucleic
acid
based
on
technology.
enables
identification
limit,
making
it
suitable
detecting
low-concentration
samples.
It
is
stable,
reliable,
exhibits
holds
potential
analysis
provides
technical
support
epidemiological
community
sewage.
shows
promising
applications
viral
monitoring.
Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown
Published: May 16, 2025
Language: Английский