Leishmaniasis
Conditions
Brief summary
This is an exploratory experimental multicentre study on archived serum samples and on prospectively collected serum samples and blood samples. General objectives of the study are: to identify specific biomarkers in order to develop more accurate serological tests for the screening of Leishmania infections that does not present cross-reaction with other infectious diseases; to unveil the Antigen Recognition Patterns (ARPs) of Leishmania infection in humans; to verify whether the ARPs and the new biomarkers are common to different Leishmania infantum strains isolated in Italy as well as in other part of the World and to different strains of L. donovani isolated in Sudan as well as in other part of the World and discard any possible cross reaction with other infectious diseases such as Chagas, TB, leprosy or malaria.
Interventions
DIAGNOSTIC_TESTIn-house Western blot, specific for Leishmania
Sample will be collected at the collaborating centres and at the DITM, if not already stored at -80°C. The samples will be delivered to the DITM. For the CONTROL groups, two serological tests for Leishmaniasis, based on different antigenic fraction, will be performed on serum samples. Different strains of Leishmania infantum and Leishmania donovani will be cultivated axenically at the DITM. Whole soluble fraction of promastigote (WSF), the whole membrane protein fraction (WMF) and the whole soluble fraction secreted by promastigotes (WSFe) of Leishmania spp. will be obtained and the ARPs from WSF, WMF and WSFe, of each Leishmania will be checked trough Western-blot analysis testing at a final dilution of 1/100 each serum sample. The predominant bands detected by Western-blot analysis will be identified by SDS-PAGE separation. Selected bands, corresponding to the immunogenic bands, will be excised, digested and identified with Mass Spectrometry analysis.
Sponsors
IRCCS Sacro Cuore Don Calabria di Negrar
Study design
Allocation
NON_RANDOMIZED
Intervention model
SINGLE_GROUP
Primary purpose
DIAGNOSTIC
Masking
NONE
Eligibility
Sex/Gender
ALL
Age
18 Years to No maximum
Healthy volunteers
Yes
Inclusion criteria
CASES: Inclusion criteria: * Leishmania positive patient from Italy (n=30) and from Sudan (n=30); * patient with a clinically or laboratory confirmed diagnosis of leishmaniasis; * signed informed consent (criteria not applied in case of samples stored at the Tropica Biobank at the DITM) * being clinically classified as leishmaniasis patient by an experienced clinician or being laboratory confirmed as leishmaniasis patients, giving positive to at least two different serological tests or to a molecular test for Leishmania sp. * adult subjects (equal or older than 18 years)
Exclusion criteria
* lack of the necessary data and/or of biological samples * subjects younger than 18 years COMMUNITY CONTROLS: Inclusion criteria: * Cryo-preserved serum samples stored at the Tropica Biobank at the DITM , IRCCS Sacro Cuore Don Calabria Hospital, Negrar (Vr), from subject from Italy (n=30) and from Africa (n=30) with no infection due by Leishmania spp. * signed informed consent to Tropica Biobank * adult subjects (equal or older than 18 years) * being negative to two different serological tests for leishmaniasis
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Biomarkers from Leishmania spp | Baseline | Presence/Absence of a reactive immunogenic band or a pattern of reactive immunogenic bands in an in-house Western blot, specific for Leishmania cases respect to negative controls. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| ARPs in different Leishmania infantum and L. donovani strains | Baseline | Presence/Absence of a common or differential reactive immunogenic band/pattern in different Leishmania infantum and Leishmania donovani strains from different geographical areas. |
| Cross reaction | Baseline | Presence/Absence of cross-reaction of the identified immunogenic bands, testing serum samples of patients with malaria or TB or leprosy or Chagas' disease |
Countries
Italy
Outcome results
None listed