Visceral Leishmaniasis Research Articles

Leishmania exploits host cAMP/EPAC/calcineurin signaling to induce an IL-33–mediated anti-inflammatory environment for the establishment of infection

Host anti-inflammatory responses are critical for the progression of visceral leishmaniasis and the pleomorphic cytokine IL-33 was found to be upregulated in infection. The underlying mechanism is not yet known. Here, we documented that IL-33 induction is a consequence of elevated cAMP-mediated EPAC/calcineurin-dependent signaling and is essential for the sustenance of infection. L. donovani-infected RAW and bone marrow-derived macrophages showed significant up-regulation of IL-33 and its neutralization by anti-IL-33 antibody resulted in decreased parasite survival and increased inflammatory responses. Infection-induced elevated cAMP was involved in IL-33 production and of its two downstream effectors PKA and EPAC, only the latter was responsible for elevated IL-33 levels. EPAC initiated Rap-dependent phospholipase C activation, which triggered the release of intracellular calcium followed by calcium/calmodulin complex formation. Screening of calmodulin-dependent enzymes affirmed the involvement of the phosphatase calcineurin in cAMP/EPAC/calcium/calmodulin signaling-induced IL-33 production and parasite survival. Activated calcineurin thereby ensured nuclear localization of the transcription factors NFATc1 and HIF-1α for IL-33 transcription and we further ascertained their role in IL-33 transcription using a ChIP assay. Administering specific inhibitors of NFATc1 and HIF-1α in a BALB/c mouse model of visceral leishmaniasis led to decreased liver and spleen parasite burden with concomitant decrease in IL-33 levels. Splenocyte supernatants of inhibitor-treated infected mice further documented an increase in TNF-α and IL-12 with simultaneous decrease of IL-10, thereby indicating an overall disease-escalating effect of IL-33. Thus, this study demonstrates that cAMP/EPAC/calcineurin signaling is crucial for the activation of IL-33 and in effect creates anti-inflammatory responses, essential for infection.

Skin pharmacokinetics of miltefosine in the treatment of post-kala-azar dermal leishmaniasis in South Asia.

Post-kala-azar dermal leishmaniasis (PKDL) arises as a dermal complication following a visceral leishmaniasis (VL) infection. Current treatment options for PKDL are unsatisfactory, and there is a knowledge gap regarding the distribution of antileishmanial compounds within human skin. The present study investigated the skin distribution of miltefosine in PKDL patients, with the aim to improve the understanding of the pharmacokinetics at the skin target site in PKDL. Fifty-two PKDL patients underwent treatment with liposomal amphotericin B (20 mg/kg) plus miltefosine (allometric dosing) for 21 days. Plasma concentrations of miltefosine were measured on study days 8, 15, 22 and 30, while a punch skin biopsy was taken on day 22. A physiologically based pharmacokinetic (PBPK) model was developed to evaluate the distribution of miltefosine into the skin. Following the allometric weight-based dosing regimen, median miltefosine concentrations on day 22 were 43.73 µg/g (IQR: 21.94-60.65 µg/g) in skin and 33.29 µg/mL (IQR: 25.9-42.58 µg/mL) in plasma. The median individual concentration ratio of skin to plasma was 1.19 (IQR: 0.79-1.9). In 87% (45/52) of patients, skin exposure was above the suggested EC90 PK target of 10.6 mg/L associated with in vitro susceptibility. Simulations indicated that the residence time of miltefosine in the skin would be more than 2-fold longer than in plasma, estimated by a mean residence time of 604 versus 266 hours, respectively. This study provides the first accurate measurements of miltefosine penetration into the skin, demonstrating substantial exposure and prolonged retention of miltefosine within the skin. These findings support the use of miltefosine in cutaneous manifestations of leishmaniasis. In combination with parasitological and clinical data, these results are critical for the future optimization of combination therapies with miltefosine in the treatment of PKDL.

Investigation into in silico and in vitro approaches for inhibitors targeting MCM10 in Leishmania donovani: a comprehensive study.

Visceral Leishmaniasis (VL), the second neglected tropical disease caused by various Leishmania species, presents a significant public health challenge due to limited treatment options and the absence of vaccines. The agent responsible for visceral leishmaniasis, also referred to as "black fever" in India, is Leishmania donovani. This study focuses on L. donovani Minichromosome maintenance 10 (LdMcm10), a crucial protein in the DNA replication machinery, as a potential therapeutic target in Leishmania therapy using in silico and in vitro approaches. We employed bioinformatics tools, molecular docking, and molecular dynamics simulations to predict potential inhibitors against the target protein. The research revealed that the target protein lacks homologues in the host, emphasizing its potential as a drug target. Ligands from the DrugBank database were screened against LdMcm10 using PyRx software. The top three compounds, namely suramin, vapreotide, and pasireotide, exhibiting the best docking scores, underwent further investigation through molecular dynamic simulation and in vitro analysis. The observed structural dynamics suggested that LdMcm10-ligand complexes maintained consistent binding throughout the 300ns simulation period, with minimal variations in their backbone. These findings suggest that these three compounds hold promise as potential lead compounds for developing new drugs against leishmaniasis. In vitro experiments also demonstrated a dose-dependent reduction in L. donovani viability for suramin, vapreotide, and pasireotide, with computed IC50 values providing quantitative metrics of their anti-leishmanial efficacy. The research offers a comprehensive understanding of LdMcm10 as a drug target and provides a foundation for further investigations and clinical exploration, ultimately advancing drug discovery strategies for leishmaniasis treatment.

A panel of recombinant Leishmania donovani cell surface and secreted proteins identifies LdBPK_323600.1 as a serological marker of symptomatic infection.

Visceral leishmaniasis is fatal if left untreated with patients often displaying mild and non-specific symptoms during the early stages of infection making accurate diagnosis important. Current methods for diagnosis require highly trained medical staff to perform highly invasive biopsies of the liver or bone marrow which pose risks to the patient. Less invasive molecular tests are available but can suffer from regional variations in their ability to accurately diagnose an infection. To identify new diagnostic markers of visceral leishmaniasis, we produced and tested a panel of 93 proteins identified from the genome of the parasite responsible for this disease. We found that the pattern of host antibody reactivity to these proteins was broadly consistent across naturally acquired infections in both human patients and dogs, as well as experimental rodent infections. We identified a new protein called LdBPK_323600.1 that could accurately diagnose visceral leishmaniasis infections in humans.

Simplified molecular diagnosis of visceral leishmaniasis: Laboratory evaluation of miniature direct-on-blood PCR nucleic acid lateral flow immunoassay.

Diagnosis of visceral leishmaniasis (VL) in resource-limited endemic regions is currently based on serological testing with rK39 immunochromatographic tests (ICTs). However, rK39 ICT frequently has suboptimal diagnostic accuracy. Furthermore, treatment monitoring and detection of VL relapses is reliant on insensitive and highly invasive tissue aspirate microscopy. Miniature direct-on-blood PCR nucleic acid lateral flow immunoassay (mini-dbPCR-NALFIA) is an innovative and user-friendly molecular tool which does not require DNA extraction and uses a lateral flow strip for result read-out. This assay could be an interesting candidate for more reliable VL diagnosis and safer test of cure at the point of care. The performance of mini-dbPCR-NALFIA for diagnosis of VL in blood was assessed in a laboratory evaluation and compared with the accuracy of rK39 ICTs Kalazar Detect in Spain and IT LEISH in East Africa. Limit of detection of mini-dbPCR-NALFIA was 650 and 500 parasites per mL of blood for Leishmania donovani and Leishmania infantum, respectively. In 146 blood samples from VL-suspected patients from Spain, mini-dbPCR-NALFIA had a sensitivity of 95.8% and specificity 97.2%, while Kalazar Detect had a sensitivity of 71.2% and specificity of 94.5%, compared to a nested PCR reference. For a sample set from 58 VL patients, 10 malaria patients and 68 healthy controls from Ethiopia and Kenya, mini-dbPCR-NALFIA had a pooled sensitivity of 87.9% and pooled specificity of 100% using quantitative PCR as reference standard. IT LEISH sensitivity and specificity in the East African samples were 87.9% and 97.4%, respectively. Mini-dbPCR-NALFIA is a promising tool for simplified molecular diagnosis of VL and follow-up of treated patients in blood samples. Future studies should evaluate its use in endemic, resource-limited settings, where mini-dbPCR-NALFIA may provide an accurate and versatile alternative to rK39 ICTs and aspirate microscopy.

Visceral Leishmaniasis-Human Immunodeficiency Virus-Coinfected Patients Are Highly Infectious to Sandflies in an Endemic Area in India.

In an area endemic with Indian visceral leishmaniasis (VL), we performed direct xenodiagnosis to evaluate the transmission of Leishmania donovani from patients with VL-human immunodeficiency virus (HIV) coinfection to the vector sandflies, Phlebotomus argentipes. Fourteen patients with confirmed VL-HIV coinfection, with a median parasitemia of 42 205 parasite genome/mL of blood, were exposed to 732 laboratory-reared pathogen-free female P argentipes sandflies on their lower arms and legs. Microscopy revealed that 16.66% (122/732) of blood-fed flies were xenodiagnosis positive. Notably, 93% (13/14) of the VL-HIV group infected the flies, as confirmed by quantitative polymerase chain reaction and/or microscopy, and were 3 times more infectious than those who had VL without HIV.

Persistent T cell unresponsiveness associated with chronic visceral leishmaniasis in HIV-coinfected patients

A large proportion of HIV-coinfected visceral leishmaniasis (VL-HIV) patients exhibit chronic disease with frequent VL recurrence. However, knowledge on immunological determinants underlying the disease course is scarce. We longitudinally profiled the circulatory cellular immunity of an Ethiopian HIV cohort that included VL developers. We show that chronic VL-HIV patients exhibit high and persistent levels of TIGIT and PD-1 on CD8+/CD8- T cells, in addition to a lower frequency of IFN-γ+ TIGIT- CD8+/CD8- T cells, suggestive of impaired T cell functionality. At single T cell transcriptome and clonal resolution, the patients show CD4+ T cell anergy, characterised by a lack of T cell activation and lymphoproliferative response. These findings suggest that PD-1 and TIGIT play a pivotal role in VL-HIV chronicity, and may be further explored for patient risk stratification. Our findings provide a strong rationale for adjunctive immunotherapy for the treatment of chronic VL-HIV patients to break the recurrent disease cycle.

A recombinant chimeric antigen constructed with B-cell epitopes from Mycobacterium leprae hypothetical proteins is effective for the diagnosis of leprosy

The diagnosis if leprosy is difficult, as it requires clinical expertise and sensitive laboratory tests. In this study, we develop a serological test for leprosy by using bioinformatics tools to identify specific B-cell epitopes from Mycobacterium leprae hypothetical proteins, which were used to construct a recombinant chimeric protein, M1. The synthetic peptides were obtained and showed good reactivity to detect leprosy patients, although the M1 chimera have showed sensitivity (Se) and specificity (Sp) values higher than 90.0% to diagnose both paucibacillary (PB) and multibacillary (MB) leprosy patients, but not those developing tegumentary or visceral leishmaniasis, tuberculosis, Chagas disease, malaria, histoplasmosis and aspergillosis, in ELISA experiments. Using sera from household contacts, values for Se and Sp were 100% and 65.3%, respectively. In conclusion, our proof-of-concept study has generated data that suggest that a new recombinant protein could be developed into a diagnostic antigen for leprosy.

Blood transfusion in the care of patients with visceral leishmaniasis: a review of practices in therapeutic efficacy studies.

Blood transfusion remains an important aspect of patient management in visceral leishmaniasis (VL). However, transfusion triggers considered are poorly understood. This review summarises the transfusion practices adopted in VL efficacy studies using the Infectious Diseases Data Observatory VL clinical trials library. Of the 160 studies (1980-2021) indexed in the IDDO VL library, description of blood transfusion was presented in 16 (10.0%) (n=3459 patients) studies. Transfusion was initiated solely based on haemoglobin (Hb) measurement in nine studies, combining Hb measurement with an additional condition (epistaxis/poor health/clinical instability) in three studies and the criteria was not mentioned in four studies. The Hb threshold range for triggering transfusion was 3-8 g/dL. The number of patients receiving transfusion was explicitly reported in 10 studies (2421 patients enrolled, 217 underwent transfusion). The median proportion of patients who received transfusion in a study was 8.0% (Interquartile range: 4.7% to 47.2%; range: 0-100%; n=10 studies). Of the 217 patients requiring transfusion, 58 occurred before VL treatment initiation, 46 during the treatment/follow-up phase and the time was not mentioned in 113. This review describes the variation in clinical practice and is an important initial step in policy/guideline development, where both the patient's Hb concentration and clinical status must be considered.

Apolipoprotein E Is Upregulated in Blood and Circulating Monocytes of Indian Patients With Visceral Leishmaniasis.

Apolipoprotein E (ApoE) has been associated with several diseases including Parkinson's disease, Alzheimer's and multiple sclerosis. ApoE also has documented immunomodulatory functions. We investigated gene expression in circulating monocytes and in bone marrows of patients with visceral leishmaniasis (VL) living in an endemic area in Bihar, India, and contrasted these with control healthy subjects or other diagnostic bone marrows from individuals in the same region. Samples from VL patients were obtained prior to initiating treatment. Our study revealed significant upregulated expression of the apoE transcript in patients with VL. Furthermore, the levels of ApoE protein were elevated in serum samples of subjects with VL compared with healthy endemic controls. These observations may provide clues regarding the complex interactions between lipid metabolism and immunoregulation of infectious and inflammatory diseases.

Effectiveness of an Intervention in Improving Knowledge and Practices about Kala-azar in a Tribal Population of Eastern India

Abstract Background: Kala-azar, or visceral leishmaniasis, caused by Leishmania donovani, remains a significant health concern in India and other endemic regions. This study aimed to assess the knowledge and practices (KAP) related to kala-azar among the Bedia tribal community in the Uttar Dinajpur district of West Bengal, India, and evaluate the impact of an educational intervention. Methods: A total of 289 participants were included in this observational study with an intervention component. Demographic and socioeconomic data were collected, and a pretested questionnaire assessed KAP related to kala-azar. Knowledge and practice assessments were conducted before and after the intervention. Statistical analysis was done to evaluate the intervention’s effectiveness. Results: The majority of participants were in the 18–30-year age group, engaged in unskilled work, and had limited formal education, reflecting significant poverty in the population. The intervention significantly improved participants’ knowledge and practices related to kala-azar. After the intervention, correct responses to questions regarding transmission, symptoms, curability, and treatment increased substantially. Discussion: The study highlights the effectiveness of targeted interventions in raising awareness and promoting healthier practices in underserved communities. It underscores the importance of addressing socioeconomic disparities in health-care access and tailoring interventions to the specific needs of different socioeconomic statuses. Conclusion: This study provides critical insights into the KAP related to kala-azar among the Bedia tribal community in India. Targeted interventions can be effective in reducing the incidence of kala-azar and improving overall health outcomes in marginalized populations.

Seroepidemiological survey for visceral leishmaniasis in dogs from an unaffected municipality of RS in the period 2021-2024

Canine visceral leishmaniasis (CVL) is a vector-borne parasitic disease that is expanding geographically, with autochthonous cases recorded in Rio Grande do Sul (RS) since 2008. Indigenous areas such as the municipalities of Pelotas, Rio Grande and Bagé are not part of active epidemiological surveillance for CVL, but the region has predisposing factors for the spread of the disease. The aim of this study was therefore to carry out a seroepidemiological survey of serum samples from dogs treated at a veterinary hospital in a disease-free area. The survey took place over a three-year period, following the current Ministry of Health (MoH) protocol. A total of 2,365 samples from asymptomatic animals were tested, of which 0.8% were seroreactive in the Dual Path Platform (DPP®) screening test. Of the samples that reacted to the DPP®, 37% were seroreactive to the ELISA (Bio-Manguinhos®), one was inconclusive (5.2%), 42% were negative and in 21% no confirmatory test was carried out. It can be concluded that Pelotas, Rio Grande and Bagé, the places of residence of seroreagent animals and CVL-free municipalities, have seroreagent dogs circulating according to the current MH protocol and require attention from epidemiological surveillance.

Immunotherapy Combining Mimotopes Selected by Phage Display Plus Amphotericin B Is Effective for Treatment Against Visceral Leishmaniasis.

The treatment for visceral leishmaniasis (VL) causes toxicity in patients, entails high cost and/or leads to the emergence of resistant strains. No human vaccine exists, and diagnosis presents problems related to the sensitivity or specificity of the tests. Here, we tested two phage clones, B1 and D11, which were shown to be protective against Leishmania infantum infection in a murine model as immunotherapeutics to treat mice infected with this parasite species. The phages were used alone or with amphotericin B (AmpB), while other mice received saline, AmpB, a wild-type phage (WTP) or WTP/AmpB. Results showed that the B1/AmpB and D11/AmpB combinations induced polarised Th1-type cellular and humoral responses, which were primed by high levels of parasite-specific IFN-γ, IL-12, TNF-α, nitrite and IgG2a antibodies, which reflected in significant reductions in the parasite load in distinct organs of the animals when analyses were performed 1 and 30 days after the treatments. Reduced organic toxicity was also found in these animals, as compared with the controls. In conclusion, preliminary data suggest the potential of the B1/AmpB and D11/AmpB combinations as immunotherapeutics against L. infantum infection.

Visceral leishmaniasis associated with macrophage activation syndrome: Case report

Visceral leishmaniasis associated with macrophage activation syndrome: Case report

Analyzing the Impact of Control Strategies on VisceralLeishmaniasis: A Mathematical Modeling Perspective

Visceral Leishmaniasis remains a significant public health challenge in eastern Sudan despite extensive control efforts. This study employs MCMC Bayesian inference techniques to fit a mathematical model for studying visceral Leishmaniasis transmission dynamics with interventions. The research focuses on evaluation of control programs to adapt to the dynamic nature of visceralLeishmaniasis transmission. We analyze 22 years of cumulative visceral Leishmaniasis cases from eastern Sudan, an endemic region for visceral Leishmaniasis, and assessed the effectiveness of interventions implemented thus far. The results reveal that visceral Leishmaniasis is prevalent, with reported cases representing less than 20% of community infections. Improved surveillance and diagnostics are necessary for accurately estimating the disease burden. The effectiveness of intervention strategies for vector control for reducing VL transmission in the region is found to be limited. Furthermore, the model predicts visceral Leishmaniasis cases will continue to increase in the near future, underscoring the need adaptable initiatives to reduce the disease burden.

A link between circulating immune complexes and acute kidney injury in human visceral leishmaniasis

Visceral leishmaniasis (VL) is an infectious disease caused by Leishmania infantum. Clinically, VL evolves with systemic impairment, immunosuppression and hyperactivation with hypergammaglobulinemia. Although renal involvement has been recognized, a dearth of understanding about the underlying mechanisms driving acute kidney injury (AKI) in VL remains. We aimed to evaluate the involvement of immunoglobulins (Igs) and immune complexes (CIC) in the occurrence of AKI in VL patients. Fourteen VL patients were evaluated between early treatment and 12 months post-treatment (mpt). Anti-Leishmania Igs, CIC, cystatin C, C3a and C5a were assessed and correlated with AKI markers. Interestingly, high levels of CIC were observed in VL patients up to 6 mpt. Concomitantly, twelve patients met the criteria for AKI, while high levels of cystatin C were observed up to 6 mpt. Plasmatic cystatin C was positively correlated with CIC and Igs. Moreover, C5a was correlated with cystatin C, CIC and Igs. We did not identify any correlation between amphotericin B use and kidney function markers in VL patients, although this association needs to be further explored in subsequent studies. Our data reinforce the presence of an important renal function impairment during VL, suggesting the involvement of Igs, CIC, and C5a in this clinical condition.

The one health approach: combating the persistent Visceral Leishmaniasis (Kala-azar) outbreaks in Isiolo County, Kenya

Background: Visceral leishmaniasis has remained a public health problem in Kenya despite Ministry of Health’s numerous efforts. This is associated with absence of a multidisciplinary response to outbreaks. We intended to analyze community’s prioritization of causes of Visceral leishmaniasis as well as community awareness on the disease prevention and control. Methods: This was a descriptive cross-sectional study design conducted between September 2023 to December 2023. 433 households’ heads, 23 community leaders and health workers were study informants. Results: Study findings showed clear proximity of human settlements and socioeconomic activities to the vector sandfly breeding sites. Environmental factors augmented by effects of climate change including flooding and high temperatures have sustained transmission including emergence of the disease in new foci. Low socio-economic status, environmental factors and climate change effects showed highly significant correlations with the disease and interplay with one another. Study results show lack of strategies and policies that promote effective prevention and control of Kala-azar. Stakeholders’ analysis revealed limited community awareness regarding roles played by different individuals in Kala-azar prevention, control, and elimination. Conclusions: There is a major gap in cooperation of partners in human, animal, and environmental health for achievements in public health strategies, therefore One Health approach should be adopted for integrated prevention, control, and elimination. The approach will sustainably improve and promote health while addressing the entire spectrum of disease control, including detection, preparedness, response, and management in addition to disease prevention averting this terrible disease from resurfacing in the future.

Modeling and analysis of visceral leishmaniasis dynamics using fractional‐order operators: A comparative study

An in‐depth understanding of the mechanism concerning the parasitic disease called visceral leishmaniasis (VL) remains challenging. Thus, we modeled the dynamics of this illness using two fractional‐order operators, including Caputo–Fabrizio and Atangana–Baleanu. In the proposed dynamical model, the endemic and disease‐free equilibrium points were considered the symmetrical components. The fractional Euler method was applied to simulate the developed model, thus determining the equilibrium points' stability. The numerical simulation results were compared with the measured data to validate the model. The results obtained from the optimum fractional operator disclosed the minimum absolute and relative errors. The primary outcome of our study is the successful application of fractional‐order operators, specifically the Atangana–Baleanu operators with = 0.98, in modeling the dynamics of VL. Notably, the numerical simulation results, validated against real data from Sudan, demonstrated that the Atangana–Baleanu operators with = 0.98 yielded the best performance, with minimum absolute and relative errors. This underscores the precision of our fractional calculus‐based dynamical model in predicting VL dynamics compared to the classical framework, particularly for fractional‐order values of = 0.99 and = 0.98.

Community Conversations: A Tool in Improving Health Outcomes and Reducing the Burden of Visceral Leishmaniasis (Kala-azar) in Isiolo County, Kenya

Visceral Leishmaniasis (Kala-azar) is a neglected tropical disease (NTD) caused by Leishmania species that affects human. Transmission is through the bite of Phlebotomus sandflies. Globally, WHO ranks Kala-azar as the second largest parasitic killer after Malaria. The disease poses a threat to more than a billion people on almost every continent. Approximately 90% of Kala-azar global burden is found in Africa. Major endemicity is documented in Eastern Africa (Kenya, South Sudan, Sudan, Ethiopia, and Somalia). Accurate burden of the disease in Kenya is unknown attributable to inadequate information on disease prevalence, diagnostic challenges, and spatial distribution. Additionally, morbidity and mortality levels from the disease in the Country are unknown ascribable to a low index of suspicion by healthcare workers, diagnostic challenges, and case management. An example of diagnostic challenges is a reported outbreak of ‘unknown disease’ akin to Kala-azar in Marsabit County in May 2023 where nine people were confirmed dead and over 80 hospitalized. This study aimed to analyze community’s prioritization of causes of Kalaazar as well as community awareness of disease prevention and control. Quantitative data were collected using Participatory Epidemiology methods in purposively selected study villages. Qualitative data were collected through semi-structured interviews. Study findings indicated that 97.2% of respondents had heard about Kala-azar. Despite the awareness, a broad gap between knowledge and practice of prevention and control strategies of the disease was evident. Kala-azar was associated with human behaviour like preferences for traditional and alternative treatment approaches (45.2%), evening outdoor practices before going to bed (99.6%), sleeping outdoors (60.1%), occasional bed nets use (69.2%), and availability of domestic animals and wild animals within house yards (54.6%). Cultural beliefs and practices were noted to play a key role in augmenting the disease burden, where most cases had burn scars from traditional healers’ therapeutic burns. As a consequence, they sought medical help after onset of complications, which ranged from increased severity to fatalities. Community engagement was minimal in prevention and control. In this participatory research, we propose behaviour change communication activities for the elimination of Kala-azar as a public health problem. This article serves as a base for future studies aimed at giving voice to communities while enhancing their understanding of causes of diseases, the possibility of enhanced early diagnostic techniques, and options for prevention, control, and surveillance which in turn is translated to action.

Synthetic Peptides Selected by Immunoinformatics as Potential Tools for the Specific Diagnosis of Canine Visceral Leishmaniasis.

Diagnosing canine visceral leishmaniasis (CVL) in Brazil faces challenges due to the limitations regarding the sensitivity and specificity of the current diagnostic protocol. Therefore, it is urgent to map new antigens or enhance the existing ones for future diagnostic techniques. Immunoinformatic tools are promising in the identification of new potential epitopes or antigen candidates. In this study, we evaluated peptides selected by epitope prediction for CVL serodiagnosis in ELISA assays. Ten B-cell epitopes were immunogenic in silico, but two peptides (peptides No. 45 and No. 48) showed the best performance in vitro. The selected peptides, both individually and in combination, were highly diagnostically accurate, with sensitivities ranging from 86.4% to 100% and with a specificity of approximately 90%. We observed that the combination of peptides showed better performance when compared to peptide alone, by detecting all asymptomatic dogs, showing lower cross-reactivity in sera from dogs with other canine infections, and did not detect vaccinated animals. Moreover, our data indicate the potential use of immunoinformatic tools associated with ELISA assays for the selection and evaluation of potential new targets, such as peptides, applied to the diagnosis of CVL.