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Published on: July 28, 2010
Immunodetection and molecular determination of visceral and cutaneous Leishmania infection using patients' urine
Asad Mirzaei1, Fereshteh Ahmadipour2, Arnaud Cannet3
1Parasitology Department, Paramedical School, Ilam university of Medical Sciences, Iran.
Insights
Urine immunoblotting offers a non-invasive method for diagnosing leishmaniasis. This study shows urine analysis is effective for detecting both cutaneous and visceral leishmaniasis, complementing traditional methods.
Area of Science:
- Medical Parasitology
- Infectious Diseases
- Diagnostic Development
Background:
- Leishmaniasis diagnosis typically requires invasive procedures for sample collection.
- Non-invasive methods using body fluids could significantly improve diagnostic accessibility.
- Urine presents a promising alternative body fluid for non-invasive leishmaniasis detection.
Purpose of the Study:
- To evaluate the efficacy of urine immunoblotting for diagnosing cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL).
- To compare the diagnostic performance of urine-based methods with established molecular techniques.
- To assess the feasibility of urine analysis as a non-invasive diagnostic tool for leishmaniasis.
Main Methods:
- Collected sera and corresponding urine samples from 37 suspected leishmaniasis patients and 3 healthy controls.
- Utilized immunoblotting (Western blot) for immunodetection of Leishmania parasites in sera and urine.
- Employed conventional and quantitative PCR for molecular confirmation and parasite load quantification.
- Sequenced PCR amplicons for precise identification of Leishmania species.
Main Results:
- Urine DNA detection rates were 86.9% for CL and 92.8% for VL.
- Immunodetection showed high positivity rates in sera (95.6% for CL, 100% for VL).
- Urine immunodetection yielded positive results in 78.2% of CL cases and 92.8% of VL cases.
- Parasite load was generally lower in urine compared to sera or tissue biopsies.
Conclusions:
- Urine immunodetection and molecular analysis are viable non-invasive tools for diagnosing CL and VL.
- Urine analysis offers a practical alternative for leishmaniasis diagnosis, reducing the need for invasive procedures.
- These findings support the use of urine-based diagnostics for leishmaniasis surveillance and patient management.
Abstract:
The diagnosis of leishmaniasis relies mainly on the use of invasive processes, to collect the biological material for detecting Leishmania parasites. Body fluids, which can be collected by non-invasive process, would greatly facilitate the leishmaniasis diagnosis. In the present study, we investigated the potency of urine immunoblotting to diagnose cutaneous and visceral leishmaniasis and we compared with routine molecular methods. A total of 80 samples, including 40 sera and their 40 corresponding urine samples were collected from 37 suspected patients with cutaneous and visceral leishmaniasis, and 3 healthy individuals (as control), in Ilam and Ardabil provinces of Iran. All sera and urine samples were analyzed, using immunoblotting. The confirmation of leishmaniasis infection was performed, using conventional and quantitative PCRs as well as by sequencing the amplicons. Among 37 suspected patients, 23 patients presented cutaneous lesions (CL) and 14 exhibited clinical symptoms reminiscent of visceral leishmaniasis (L. infantum). Among cutaneous patients, 15 were positive for zoonotic cutaneous leishmaniasis (L. major), and eight for anthroponotic cutaneous leishmaniasis (L. tropica). Molecular quantification of Leishmania parasites was performed on sera, urines and cutaneous biopsies of CL and VL patients, demonstrating that parasite load is lower in urines, compared to sera or biopsy. DNA can be detected in 20 out of 23 (86.9%) CL urine samples and in 13 out of 14 (92.8%) VL urine samples. Immunodetection analysis demonstrates that 22 out of 23 (95.6%) sera from CL patients and all patients suspected with VL are positive. For urine samples, 18 out of 23 (78.2%) urine of CL patients and 13 out of 14 (92.8%) urine of VL patients were positive, using Western blot. Therefore, immunodetection and molecular analysis using urine samples can be used as a diagnostic tool for surveying cutaneous and visceral leishmaniasis.
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