Development of a duplex real-time recombinase aided amplification assay for the simultaneous and rapid detection of PCV3 and PCV4
Renjie Sun,
Hanze Liu,
Siqi Sun
и другие.
Virology Journal,
Год журнала:
2025,
Номер
22(1)
Опубликована: Фев. 1, 2025
Porcine
circoviruses
3
(PCV3)
and
4
(PCV4)
are
emerging
pathogens
with
global
implications
for
swine
industry,
disturbing
the
diagnosis
of
PCVs
associated
diseases
due
to
a
range
similar
clinical
symptoms
increasingly
coinfections.
A
rapid
accurate
method
detection
PCV3
PCV4
is
critical
controlling
transmission
disease.
We
developed
duplex
real-time
recombinase
aided
amplification
(RAA)
assay
both
simultaneously.
The
was
completed
within
20
min
at
39℃
designed
optimal
primers
probes.
established
more
convenient
simpler
operation
compared
conventional
molecular
biological
assays.
achieved
limit
73.67
copies/reaction
each
circovirus
(at
95%
probability
by
probit
regression
analysis)
showed
high
specificity
no
cross-reactivity
other
important
porcine
viruses
(including
PCV2).
intra-
inter-group
coefficients
variation
(CV)
were
ranged
from
2.08
4.97%,
indicating
stability
reliability.
Comparative
analysis
qPCR
on
60
samples
artificially
spiked
indicated
congruence
(the
kappa
value
0.966
1,
respectively,
p
<
0.001),
only
minor
discrepancies,
validating
effectiveness
RAA
in
detecting
co-infections
its
suitability
preliminary
PCV4.
This
study
provides
robust
basis
multiplex
veterinary
using
technique,
enhancing
field's
capacity
control
PCV4,
supporting
reliable
aid
epidemiological
understanding
circoviruses.
Язык: Английский
First molecular detection and genetic characterization of porcine circovirus 4 in the Gansu Province of China
PLoS ONE,
Год журнала:
2024,
Номер
19(2), С. e0293135 - e0293135
Опубликована: Фев. 5, 2024
Since
its
initial
discovery
in
the
Hunan
province
of
China,
genomic
DNA
porcine
circovirus
4
(PCV4)
has
been
detected
pigs
across
multiple
provinces
as
well
South
Korea.
However,
prevalence
type
Gansu
Province,
remains
unknown.
To
address
this
gap,
we
undertook
an
extensive
study
where
gathered
121
clinical
samples
displaying
diverse
manifestations
from
pig
farms
Province
between
2022
and
2023.
Employing
a
real-time
fluorescence
quantification
method,
identified
presence
PCV4
genome.
Out
analyzed,
13
tested
positive
for
PCV4,
resulting
rate
10.74%
(13/121).
This
finding
confirms
within
China.
Furthermore,
successfully
sequenced
analyzed
complete
genomes
two
distinct
strains,
comparing
them
with
60
reference
sequences
archived
GenBank
database.
The
results
revealed
high
nucleotide
homology
(98.2–98.8%)
strains
obtained
indicating
relatively
low
evolutionary
Phylogenetic
analysis
that
belong
to
PCV4a
PCV4c.
As
far
know,
marks
inaugural
report
on
molecular
identification
attributes
offering
valuable
insights
devising
preventive
control
strategies
against
emerging
virus.
Язык: Английский
Generation of porcine circovirus type 4 virus-like particles and their use to detect serum antibodies
Archives of Virology,
Год журнала:
2024,
Номер
169(3)
Опубликована: Март 1, 2024
Язык: Английский
Molecular detection and genetic characteristics of porcine circovirus 3 and porcine circovirus 4 in central China
Archives of Virology,
Год журнала:
2024,
Номер
169(5)
Опубликована: Май 1, 2024
Язык: Английский
The Nuclear Localization Signal of Porcine Circovirus Type 4 Affects the Subcellular Localization of the Virus Capsid and the Production of Virus-like Particles
International Journal of Molecular Sciences,
Год журнала:
2024,
Номер
25(5), С. 2459 - 2459
Опубликована: Фев. 20, 2024
Porcine
circovirus
4
(PCV4)
is
a
newly
identified
virus
belonging
to
PCV
of
the
Circoviridae
family,
Circovirus
genus.
We
previously
found
that
PCV4
pathogenic
in
vitro,
while
virus’s
replication
cells
still
unknown.
In
this
study,
we
evaluated
N-terminal
capsid
(Cap)
and
an
NLS
at
amino
acid
residues
4–37
N-terminus
Cap,
4RSRYSRRRRNRRNQRRRGLWPRASRRRYRWRRKN37.
The
was
further
divided
into
two
fragments
(NLS-A
NLS-B)
based
on
predicted
structure,
including
α-helixes,
which
were
located
4RSRYSRRRRNRRNQRR19
24PRASRRRYRWRRK36,
respectively.
Further
studies
showed
NLS,
especially
first
α-helixes
formed
by
NLS-A
fragment,
determined
nuclear
localization
Cap
protein,
4RSRY7
critical
motif
affecting
VLP
packaging.
These
results
will
provide
theoretical
basis
for
elucidating
infection
mechanism
developing
subunit
vaccines
VLPs.
Язык: Английский
First report on identification and genomic analysis of a novel porcine circovirus (porcine circovirus 4) in cats
Frontiers in Microbiology,
Год журнала:
2023,
Номер
14
Опубликована: Сен. 22, 2023
Porcine
circovirus
type
4
(PCV4)
is
an
emerging
circovirus,
which
has
been
detected
in
domestic
pigs
across
various
provinces
China
and
Korea.
In
this
study,
we
aimed
to
investigate
whether
cats
are
susceptible
PCV4.
For
purpose,
collected
116
cat
samples
from
animal
hospitals
Sichuan
Province,
China,
between
2021
2022.
Using
a
SYBR
Green-based
real-time
PCR
assay,
PCV4
5
out
of
the
clinical
samples,
indicating
positive
rate
4.31%
(5/116)
confirming
presence
China.
Moreover,
successfully
sequenced
analyzed
complete
genome
one
strain
(SCGA-Cat)
along
with
60
reference
sequences
deposited
GenBank
database.
SCGA-Cat
exhibited
high
nucleotide
homology
(98.2-99.0%)
strains
other
species,
including
dogs,
pigs,
dairy
cows,
fur
animals.
Notably,
clustered
closely
derived
pig
Fujian
To
best
our
knowledge,
study
represents
first
report
on
molecular
detection
worldwide,
prompted
us
understand
genetic
diversity
cross-species
transmission
ongoing
cases.
However,
further
investigations
needed
explore
association
infection
syndromes
cats.
Язык: Английский
UNVEILING THE GENOMIC DIVERSITY OF PORCINE CIRCOVIRUS 4 BY BIOINFORMATICS ANALYSIS
Global Journal For Research Analysis,
Год журнала:
2024,
Номер
unknown, С. 82 - 83
Опубликована: Апрель 15, 2024
Porcine
Circovirus
4
(PCV4)
is
the
latest
type
of
Circoviruses
(PCVs)
and
found
to
be
associated
with
several
clinical
manifestations
in
pigs,
although
exact
pathogenesis
still
unclear.
This
study
aims
reveal
genetic
diversity
different
strains
PCV4
that
were
detected
locations.
Accordingly,
seventy
numbers
complete
genome
13
sequences
other
PCVs
retrieved
from
public
database
phylogenetic
analysis,
pairwise
identity
intergenomic
analysis
done
online
offline
bioinformatics
tools.
Phylogenetic
showed
formed
a
separate
clade
variation
among
between
0.0-2.3%.
The
presented
this
will
helpful
conduct
future
research
designing
molecular
diagnostics
for
detection
PCV4,
however,
more
detailed
required
better
genomic
characterization
virus.
Язык: Английский
Generation of porcine circovirus type 4 virus-like particles and their use to detect serum antibodies
Research Square (Research Square),
Год журнала:
2023,
Номер
unknown
Опубликована: Авг. 11, 2023
Abstract
Porcine
Circovirus
type
4
(PCV4),
first
identified
in
2019
as
a
newly
emerging
pathogen,
has
been
found
several
provinces
of
China,
well
Korea
and
Thailand.
Since
PCV4
is
not
included
immunization
programs,
epidemiological
investigations
should
be
conducted
for
PCV4-positive
antibodies
detection.
Virus-like
particles
(VLPs)
are
commonly
employed
serological
analysis
pathogen
infections.
However,
there
no
reports
on
using
VLPs
infection
investigation.
In
this
study,
we
successfully
generated
self-assembled
an
E.coli
expression
system
purified
the
through
two-step
purification
process.
Subsequently,
utilized
encapsulated
antigens
to
develop
indirect
ELISA.
The
established
ELISA
method
showed
high
specificity,
sensitivity,
repeatability,
making
it
suitable
investigation
serum
samples.
Finally,
was
applied
detect
422
samples
from
regions
which
134
tested
positive.
Therefore,
PCV4-VLPs-based
could
effectively
against
samples,
contributing
better
understanding
epidemiology.
Язык: Английский