Potential application of rLc36 protein for diagnosis of canine visceral leishmaniasis

Abstract

Visceral leishmaniasis (VL) is fatal if left untreated. Infected dogs are important reservoirs of the disease, and thus specific identification of infected animals is very important. Several diagnostic tests have been developed for canine VL (CVL); however, these tests show varied specificity and sensitivity. The present study describes the recombinant protein rLc36, expressed by Leishmania infantum, as potential antigen for more sensitive and specific diagnosis of CVL based on an immunoenzymatic assay. The concentration of 1.0 μg/mL of rLc36 enabled differentiation of positive and negative sera and showed a sensitivity of 85% and specificity of 71% (with 95% confidence), with an accuracy of 76%.

Improving the reference standard for the diagnosis of canine visceral leishmaniasis: a challenge for current and future tests

“…In fact, mathematical modelling indicates that the key to the potential success of CVL control may be the identification of asymptomatic dogs. (19,20) In that sense, new diagnostic tests are under development with promising results in their initial stages, (21,22,23,24) but however, it is necessary to challenge these against a robust reference standard. The current context is characterised by a lack of consensus about the best reference standard for the validation of diagnostic tests for CVL, and few studies have used reasonably adequate reference standards.…”

Alterações hematológicas em cães soropositivos ao DPP e ELISA para Leishmania infantum submetidos ao tratamento com Miltefosina 2% / Hematological alterations in dogs seropositive to DPP and ELISA for Leishmania infantum submitted to treatment with Miltefosine 2%

“…Trypanosomatidae, e transmitidas, por flebotomíneos do gênero Lutzomyia (BRASIL, 2017). A leishmaniose visceral é considerada a forma mais grave entre as leishmanioses (NOGUEIRA et al, 2018) e se encontra em franca expansão como crescente número de vetores, e de cães, principais reservatórios da LV (FERROGLIO et al, 2018;SANTOS et al, 2018). Neste cenário de distribuição o Brasil concentra 14% (3.336) dos focos eco epidemiológicos, colocando o país em destaque.…”

Overcoming challenges for amplified expression of recombinant proteins using Escherichia coli

“…The expressed recombinant P4 molecule (33 kDa) was also purified by His 6 -tag using nickel and the nitrilotriacetic acid (NTA) column, demonstrating that this strategy is efficient for protein expression and purification (Table 2). Additionally Nogueira (2014) and Nogueira et al (2018) expressed an antigen rLc36 of unknown function containing a nucleotidyl transferase important for protecting Leishmania amastigotes against reactive oxygen species produced by macrophages (Nogueira et al 2018). According to Nogueira (2014), it was possible to express rLc36 using the vector pET 28a(+) and the strain E. coli BL21 (DE) (Table 1) in order to obtain a protein with 28.4 kDa (Table 2).…”

“…Additionally Nogueira (2014) and Nogueira et al (2018) expressed an antigen rLc36 of unknown function containing a nucleotidyl transferase important for protecting Leishmania amastigotes against reactive oxygen species produced by macrophages (Nogueira et al 2018). According to Nogueira (2014), it was possible to express rLc36 using the vector pET 28a(+) and the strain E. coli BL21 (DE) (Table 1) in order to obtain a protein with 28.4 kDa (Table 2).…”