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Specific diagnosis of Opisthorchis viverrini using loop mediated isothermal amplification (LAMP) targeting parasite microsatellites.
[cholangiocarcinoma]
Opisthorchis
viverrini
and
other
food-borne
trematode
infections
are
major
health
problems
in
Thailand
,
the
Lao
People
's
Democratic
Republic
,
Vietnam
and
Cambodia
.
Differential
diagnosis
of
O
.
viverrini
based
on
the
microscopic
observation
of
parasite
eggs
is
difficult
in
areas
where
Clonorchis
sinensis
and
minute
intestinal
flukes
coexist
.
Recently
,
loop-mediated
isothermal
amplification
(
LAMP
)
has
been
widely
used
for
detection
and
identification
of
trematode
for
its
simple
method
that
is
useful
in
low
-resource
or
field
settings
.
We
have
reported
ITS
1
-
LAMP
assay
to
detect
O
.
viverrini
infection
from
human
feces
.
The
sensitivity
and
specificity
of
the
test
was
100
%
and
61
.
5
%
.
The
sensitivity
of
the
test
appeared
to
be
higher
than
microscopic
egg
examination
;
however
non-
specific
amplification
from
other
parasites
could
not
be
ruled
out
.
We
therefore
targeted
microsatellites
of
O
.
viverrini
that
is
a
species
specific
sequence
.
By
using
hydroxyl
naphthol
blue
(
HNB
)
-
LAMP
,
O
.
viverrini
microsatellite
6
(
OVMS
6
)
could
specifically
amplify
DNA
from
O
.
viverrini
genome
,
but
not
other
parasites
such
as
C
.
sinensis
,
Opisthorchis
felineus
,
Centrocestus
caninus
,
Haplorchis
taichui
,
Fasciola
gigantica
and
Haplorchoodes
sp
.
The
detection
limit
of
the
test
is
1
ng
genomic
DNA
,
which
was
1000
times
lower
than
the
ITS
1
-
LAMP
,
but
targeting
microstellites
showed
more
specific
detection
of
O
.
viverrini
.
In
addition
,
the
colorimetric
LAMP
assay
was
simple
and
effective
;
this
makes
it
potentially
applicable
for
point-of-care
diagnosis
.
Diseases
Validation
Diseases presenting
"not other parasites"
symptom
cholangiocarcinoma
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