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The performance of four molecular methods for the laboratory diagnosis of congenital toxoplasmosis in amniotic fluid samples.
[congenital toxoplasmosis]
Introduction
:
Toxoplasmosis
may
be
life-threatening
in
fetuses
and
in
immune-
deficient
patients
.
Conventional
laboratory
diagnosis
of
toxoplasmosis
is
based
on
the
presence
of
IgM
and
IgG
anti-
Toxoplasma
gondii
antibodies
;
however
,
molecular
techniques
have
emerged
as
alternative
tools
due
to
their
increased
sensitivity
.
The
aim
of
this
study
was
to
compare
the
performance
of
4
PCR-based
methods
for
the
laboratory
diagnosis
of
toxoplasmosis
.
One
hundred
pregnant
women
who
seroconverted
during
pregnancy
were
included
in
the
study
.
The
definition
of
cases
was
based
on
a
12
-
month
follow-up
of
the
infants
.
Methods
:
Amniotic
fluid
samples
were
submitted
to
DNA
extraction
and
amplification
by
the
following
4
Toxoplasma
techniques
performed
with
parasite
B
1
gene
primers
:
conventional
PCR
,
nested-
PCR
,
multiplex-nested-
PCR
,
and
real-time
PCR
.
Seven
parameters
were
analyzed
,
sensitivity
(
Se
)
,
specificity
(
Sp
)
,
positive
predictive
value
(
PPV
)
,
negative
predictive
value
(
NPV
)
,
positive
likelihood
ratio
(
PLR
)
,
negative
likelihood
ratio
(
NLR
)
and
efficiency
(
Ef
)
.
Results
:
Fifty
-
nine
of
the
100
infants
had
toxoplasmosis
;
42
(
71
.
2
%
)
had
IgM
antibodies
at
birth
but
were
asymptomatic
,
and
the
remaining
17
cases
had
non-detectable
IgM
antibodies
but
high
IgG
antibody
titers
that
were
associated
with
retinochoroiditis
in
8
(
13
.
5
%
)
cases
,
abnormal
cranial
ultrasound
in
5
(
8
.
5
%
)
cases
,
and
signs
/
symptoms
suggestive
of
infection
in
4
(
6
.
8
%
)
cases
.
The
conventional
PCR
assay
detected
50
cases
(
9
false-negatives
)
,
nested-
PCR
detected
58
cases
(
1
false-negative
and
4
false-positives
)
,
multiplex-nested-
PCR
detected
57
cases
(
2
false-negatives
)
,
and
real-time-
PCR
detected
58
cases
(
1
false-negative
)
.
Conclusions
:
The
real-time
PCR
assay
was
the
best-performing
technique
based
on
the
parameters
of
Se
(
98
.
3
%
)
,
Sp
(
100
%
)
,
PPV
(
100
%
)
,
NPV
(
97
.
6
%
)
,
PLR
(
â
ˆ
ž
)
,
NLR
(
0
.
017
)
,
and
Ef
(
99
%
)
.
Diseases
Validation
Diseases presenting
"abnormal cranial ultrasound"
symptom
congenital toxoplasmosis
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