RSC Advances,
Journal Year:
2024,
Volume and Issue:
14(21), P. 14775 - 14783
Published: Jan. 1, 2024
The
global
outbreak
of
monkeypox
virus
(MPXV)
has
highlighted
the
need
for
rapid
molecular
diagnostics
techniques.
In
this
study,
a
single-step
recombinase
polymerase
amplification
(RPA)-CRISPR/Cas12a
system
was
developed
and
sensitive
detection
MPXV.
limit
assay
1
copy
per
μL
extracted
nucleic
acids.
A
heating
lysis
method
integrated
to
further
simplify
sample
processing
workflow
shorten
time
40
min
from
result.
reaction
mixture
can
be
lyophilized
improve
its
accessibility
in
resource-limited
settings.
analysis
results
proposed
RPA-CRISPR/Cas12a
clinical
MPXV
positive
negative
samples
were
100%
consistent
with
standard
PCR
assay.
These
demonstrate
feasibility
efficiency
accurate
real-world
settings,
showcasing
potential
utility
urgent
practical
Journal of Medical Virology,
Journal Year:
2023,
Volume and Issue:
95(8)
Published: July 29, 2023
Abstract
Mpox
virus,
a
member
of
genus
Orthopoxvirus
,
causes
rash
and
flu‐like
symptoms
in
humans.
In
the
recent
global
outbreak,
it
was
reported
from
several
geographical
areas
that
have
not
historically
mpox.
Point
care,
sensitive
specific
mpox
diagnostic
assays
are
critical
checking
spread
disease.
We
developed
clustered
regularly
interspaced
short
palindromic
repeats
associated
Cas12a
nuclease‐based
assay
for
detecting
virus.
conserved
sequences
were
identified
polA
(E9L)
gene
which
differ
by
single
nucleotide
polymorphism
(SNP)
all
viruses
present
.
This
SNP
exploited
our
to
specifically
distinguish
virus
other
related
orthopox
with
limit
detection
1
copy/μl
30
min.
The
exhibits
can
prove
be
practical
value
its
surveillance
infected
multiple
viruses,
especially
hotspots
infections.
Scientific Reports,
Journal Year:
2024,
Volume and Issue:
14(1)
Published: April 3, 2024
Abstract
Anaplasma
marginale
infection
is
one
of
the
most
common
tick-borne
diseases,
causing
a
substantial
loss
in
beef
and
dairy
production
industries.
Once
infected,
pathogen
remains
cattle
for
life,
allowing
parasites
to
spread
healthy
animals.
Since
clinical
manifestations
anaplasmosis
occur
late
disease,
sensitive,
accurate,
affordable
identification
crucial
preventing
controlling
infection.
To
this
end,
we
developed
an
RPA-CRISPR/Cas12a
assay
specific
A.
bovines
targeting
msp4
gene.
Our
performed
at
moderately
high
temperature,
producing
fluorescent
signals
or
positive
readout
lateral
flow
dipstick,
which
as
sensitive
conventional
PCR-based
DNA
amplification.
This
can
detect
few
4
copies/μl
using
marker
without
cross-reactivity
other
bovine
pathogens.
Lyophilized
components
be
stored
room
temperature
extended
period,
indicating
its
potential
field
diagnosis
low-resource
settings
bovines.
RSC Advances,
Journal Year:
2024,
Volume and Issue:
14(21), P. 14775 - 14783
Published: Jan. 1, 2024
The
global
outbreak
of
monkeypox
virus
(MPXV)
has
highlighted
the
need
for
rapid
molecular
diagnostics
techniques.
In
this
study,
a
single-step
recombinase
polymerase
amplification
(RPA)-CRISPR/Cas12a
system
was
developed
and
sensitive
detection
MPXV.
limit
assay
1
copy
per
μL
extracted
nucleic
acids.
A
heating
lysis
method
integrated
to
further
simplify
sample
processing
workflow
shorten
time
40
min
from
result.
reaction
mixture
can
be
lyophilized
improve
its
accessibility
in
resource-limited
settings.
analysis
results
proposed
RPA-CRISPR/Cas12a
clinical
MPXV
positive
negative
samples
were
100%
consistent
with
standard
PCR
assay.
These
demonstrate
feasibility
efficiency
accurate
real-world
settings,
showcasing
potential
utility
urgent
practical
