Rapid detection of human and canine visceral leishmaniasis: Assessment of a latex agglutination test based on the A2 antigen from amastigote forms of Leishmania infantum

Akhoundi, Behnaz; Mohebali, Mehdi; Shojaee, Saeedeh; Jalali, M; Kazemi, Bahram; Bandehpour, Mojgan; Keshavarz, Hossein; Edrissian, Gholam Hossein; Eslami, Mohammad H.; Malekafzali, Hossein; Kouchaki, Ameneh

doi:10.1016/j.exppara.2012.12.002

Cited by 34 publications

(18 citation statements)

References 26 publications

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“…Similar to our results, Reis et al [34] showed 12.5% of false-negative results when testing ELISA-k39, due to the presence of dogs with antibody titers lower than 1∶320. Other authors [14] have showed 15% of false-negative results also due to low antibody titers, in a latex agglutination test with the A2 antigen.…”

Section: Discussionmentioning

confidence: 92%

Novel Recombinant Multiepitope Proteins for the Diagnosis of Asymptomatic Leishmania infantum-Infected Dogs

et al. 2015

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BackgroundVisceral leishmaniasis is the most severe form of leishmaniasis. Worldwide, approximately 20% of zoonotic human visceral leishmaniasis is caused by Leishmania infantum, also known as Leishmania chagasi in Latin America. Current diagnostic methods are not accurate enough to identify Leishmania-infected animals and may compromise the effectiveness of disease control. Therefore, we aimed to produce and test two recombinant multiepitope proteins as a means to improve and increase accuracy in the diagnosis of canine visceral leishmaniasis (CVL).Methodology/Principal FindingsTen antigenic peptides were identified by CVL ELISA in previous work. In the current proposal, the coding sequences of these ten peptides were assembled into a synthetic gene. Furthermore, other twenty peptides were selected from work by our group where good B and T cell epitopes were mapped. The coding sequences of these peptides were also assembled into a synthetic gene. Both genes have been cloned and expressed in Escherichia coli, producing two multiepitope recombinant proteins, PQ10 and PQ20. These antigens have been used in CVL ELISA and were able to identify asymptomatic dogs (80%) more effectively than EIE-LVC kit, produced by Bio-Manguinhos (0%) and DPP kit (10%). Moreover, our recombinant proteins presented an early detection (before PCR) of infected dogs, with positivities ranging from 23% to 65%, depending on the phase of infection in which sera were acquired.Conclusions/SignificanceOur study shows that ELISA using the multiepitope proteins PQ10 and PQ20 has great potential in early CVL diagnosis. The use of these proteins in other methodologies, such as immunochromatographic tests, could be beneficial mainly for the detection of asymptomatic dogs.

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Section: Discussionmentioning

confidence: 92%

Novel Recombinant Multiepitope Proteins for the Diagnosis of Asymptomatic Leishmania infantum-Infected Dogs

et al. 2015

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“…There are techniques such as Western blotting that is highly accurate but not available in routine practice, while others have been proposed but are not extensively used, such as the latex agglutination test or detection of antibodies through immunosensors or flow cytometry. [120][121][122][123] The most common techniques used to detect antileishmanial antibodies are based on 3 analytic principles: immunofluorescent antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA), and immunochromatographic test (ICT). The latter method is the basis of all rapid in-clinic assays, which only provide a qualitative result (ie, presence/absence of specific reactive bands or spots).…”

Section: Methodsmentioning

confidence: 99%

Laboratory tests for diagnosing and monitoring canine leishmaniasis

Paltrinieri

Gradoni

Roura

et al. 2016

Veterinary Clinical Pathol

149

232

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Although several reviews on canine leishmaniasis have been published, none thoroughly described clinicopathologic abnormalities and their clinical usefulness. The aim of this review was to provide information concerning current diagnostic tests relevant for clinical pathologists and from a practical perspective. Specifically, in canine leishmaniasis, nonregenerative normocytic normochromic anemia, thrombocytopenia, or leukogram changes may be present. Clinical chemistry and urinalysis may indicate renal dysfunction (azotemia, decreased urine specific gravity, proteinuria) and an inflammatory/immune response (increased acute phase proteins [APP] or α - and/or γ-globulins). Although a potential gammopathy is usually polyclonal, it may also appear oligo- or monoclonal, especially in dogs coinfected by other vector-borne pathogens. When lesions are accessible to fine-needle aspiration (lymphoadenomegaly, nodular lesions, joint swelling), cytology is strongly advised, as the presence of Leishmania amastigotes in a pattern of pyogranulomatous inflammation or lymphoplasmacytic hyperplasia is diagnostic. If the cytologic pattern is inconclusive, the parasite should be identified by histology/immunohistochemistry or PCR on surgical biopsies. Alternatively, cytology and PCR may be performed on bone marrow samples where amastigotes, along with erythroid hypoplasia, myeloid hyperplasia, plasmacytosis, or secondary dysmyelopoiesis can be observed. Dogs with overt leishmaniasis generally have high antibody titers, while low titers predominate in immunologically resistant infected dogs or in exposed dogs with no parasite confirmation. Quantitative serology is recommended in clinically suspect dogs as high-titer antibodies titers may confirm the clinical diagnosis. In confirmed and treated dogs, renal function and inflammatory/immune response variables should be periodically monitored.

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“…By binding either an antigen [62], [63] or a specific antibody [64], [65] of the causative agent to a latex bead, a cheap and fast screening tool for serum can be developed that minimally requires the use of any equipment. Latex bead assays have been developed for other tropical infections, including visceral leismaniasis [62], [64] and paracoccidioidomycosis [63]. In order to develop a specific agglutination assay, a discriminatory antigen needs to be selected.…”

Section: Development Of Methods For Early Diagnosismentioning

confidence: 99%

The Mycetoma Knowledge Gap: Identification of Research Priorities

et al. 2014

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Mycetoma is a tropical disease which is caused by a taxonomically diverse range of actinomycetes (actinomycetoma) and fungi (eumycetoma). The disease was only recently listed by the World Health Organization (WHO) as a neglected tropical disease (NTD). This recognition is the direct result of a meeting held in Geneva on February 1, 2013, in which experts on the disease from around the world met to identify the key research priorities needed to combat mycetoma. The areas that need to be addressed are highlighted here. The initial priority is to establish the incidence and prevalence of the disease in regions where mycetoma is endemic, prior to determining the primary reservoirs of the predominant causal agents and their mode of transmission to susceptible individuals in order to establish novel interventions that will reduce the impact of the disease on individuals, families, and communities. Critically, economical, reliable, and effective methods are required to achieve early diagnosis of infections and consequential improved therapeutic outcomes. Molecular techniques and serological assays were considered the most promising in the development of novel diagnostic tools to be used in endemic settings. Improved strategies for treating eumycetoma and actinomycetoma are also considered.

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Rapid detection of human and canine visceral leishmaniasis: Assessment of a latex agglutination test based on the A2 antigen from amastigote forms of Leishmania infantum

Cited by 34 publications

References 26 publications

Novel Recombinant Multiepitope Proteins for the Diagnosis of Asymptomatic Leishmania infantum-Infected Dogs

Novel Recombinant Multiepitope Proteins for the Diagnosis of Asymptomatic Leishmania infantum-Infected Dogs

Laboratory tests for diagnosing and monitoring canine leishmaniasis

The Mycetoma Knowledge Gap: Identification of Research Priorities

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