Diagnostic Accuracy of the Diagnostic Accuracy of the Leishmania Leishmania OligoC OligoC ‐ ‐ TesT TesT for the Diagnosis of for the Diagnosis of Cutaneous Cutaneous Leishmaniasis Leishmaniasis in Peru in Peru Diego ESPINOSA, Andrea K. BOGGILD, Stijn DEBORGGRAEVE, et al. www.corisbio.com 57 th ASTMH annual meeting – New Orleans, LA December 11, 2008
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Diagnostic Accuracy of the Diagnostic Accuracy of the LeishmaniaLeishmania OligoCOligoC‐‐TesTTesT
for the Diagnosis of for the Diagnosis of
CutaneousCutaneous
LeishmaniasisLeishmaniasis
in Peruin PeruDiego ESPINOSA, Andrea K. BOGGILD, Stijn
DEBORGGRAEVE, et al.
www.corisbio.com
57th
ASTMH annual meeting – New Orleans, LA
December 11, 2008
1 Introduction.
• Leishmaniasis
is
a
vector‐borne
disease
caused
by
obligate
intra‐macrophage
protozoan
parasites
that
belong
to
the
genus Leishmania.
• Cutaneous
leishmaniasis
(CL) results from the replication of
parasites in macrophages in the skin.
1 Introduction.
• Routine
and
definitive
diagnosis
of
CL
is
still
based
on
the
demonstration
of
amastigotes
in
skin
lesions
using
direct
smears
(a)
or
subsequent
in
vitro
cultures
(b).
These
methods, however, have several drawbacks.
(b)(a)
1 Introduction.
• The
polymerase
chain
reaction
(PCR)
is
a
useful
tool
for
the
detection of Leishmania
parasites in clinical samples.
• There is actually a demand for “low‐tech”
PCR approaches, more
suitable
for
countries
with
developing
economies
Leishmania
OligoC‐TesT.
2 Methods.
a. Diagnostic accuracy of the Leishmania
OligoC‐TesT.
– Assessed on aspirate and scraping DNA extracts of 61 lesions
from 45 CL suspected patients.
– The results of the Leishmania
OligoC‐TesT
were compared to:
• Traditional diagnostic tools (direct smears and in vitro
cultures).
• Leishmanin
Skin Test.
• kDNA
PCR.
2 Methods.
b. The Leishmania
OligoC‐TesT
in the Peruvian jungle.
Pangoa
Kiteni
2 Methods.
b. The Leishmania
OligoC‐TesT
in the Peruvian jungle.
BA
A.
Front view
of
Kinteni´s local hospital.
B.
Laboratory
where
the
assays
took
place.
2 Methods.
• The Leishmania
OligoC‐TesT
in the Peruvian Jungle.
A
B3 hours 10 minutes
TOTAL TIME FOR RESULTS
5 hours~
A
(sampling)
= 20 minutesB (DNA extraction)
= 40 minutes +
1 hour
PCR Products´ detection
3 Results.
a.
Diagnostic accuracy of the Leishmania
OligoC‐TesT.
CL = cutaneous
leishmaniasis;
95% CI = 95% confidence
interval
scored
by Wilson´s
method;
POS = positive
b.
The Leishmania
OligoC‐TesT
in the Peruvian jungle.
Decision
making
alternatives
for
diagnosis.
CULTUREDIRECT SMEAR
Sensitivity
~ 60%
Results
~ 30 minutes
Sensitivity
~ 60%
Results
~ days to weeks
and
/ or
Leishmania
OligoC‐TesT
Sensitivity
~ 90%
Results
~ 5 hours
4 Conclusions.
• When
performed
on
lesion
scrapings,
the
Leishmania
OligC‐TesT´s
sensitivity
was
higher
than
on
lesion
aspirates.
• The
Leishmania
OligoC‐TesT
can
be
conducted
in
low‐
level‐equipped laboratories.
• This
tool
is
useful
for
the
diagnosis
of
CL
in
different
experimental settings and health facilities.
Acknowledgements.•
Commission
of
the
European
communities ‐
Sixth
Framework
Programme,
priority INCO‐DEV, project TRYLEIDIAG, contract 015379.
•
Consejo
Nacional
de
Ciencia,
Tecnología
e
Innovación
Tecnológica
(CONCYTEC).
•
Corio
BioConcept.
•
Directorate
General for
Development
Cooperation
of
the
Belgian
Government
(FA2, project
95501; FA3, project
095502).
•
European Community
(INCO‐Dev
program, “Control Strategies
for
the
visceral
leishmaniasis
(VL)
and
mucocutaneous
leishmaniasis
in
South
America:
applications
of
molecular epidemiology”
[contract
INCO‐CT2005‐015407]).
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