Real-time PCR using high-resolution melting analysis technology for diagnosis of Leishmania and determination of types of clinical samples
Background/aim:
Leishmaniasis is a disease group carried by
Phlebotomus
and
Lutzomyia
sand flies infected with
Leishmania
and is
mostly observed in rural areas. In this study, using high-resolution melting analysis (HRMA), we aim to identify the active types of
leishmaniasis, which are inadequately identified by classical methods.
Materials and methods:
Samples of 85 patients were examined in the study. Six of the patients were suspected of having visceral
leishmaniasis (VL) and 79 cutaneous leishmaniasis (CL). The slides prepared from the samples were stained with Giemsa stain and
examined under a light microscope. The results were compared with those determined in real-time PCR. When the real-time PCR result
was positive for
Leishmania
, we determined the type by HRMA.
Results:
Among 85
Leishmania
amastigote samples, 28 (32.9%) of them were detected and accepted as positive by microscopic
examination. On the other hand, 25 (29.4%) of the 85 samples were found as positive using real-time PCR. In addition, when 25
Leishmania
-positive samples were examined by HRMA, the results showed that 21 (84%) were
L. tropica
, 3 (12%) were
L. major
, and 1
(4%) was
L. infantum
.
Conclusion:
Based on our findings, we conclude that using real-time PCR and HRMA is useful in determining the cause of the disease
in leishmaniasis.
___
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