The efficacy of recombinant protein lbk39 for the diagnosis of leishmaniosis in dogs

Abstract

Visceral leishmaniosis is one of the most important zoonotic diseases on the planet and dogs are the main reservoir of canine visceral leishmaniosis (CVL) in endemic areas. They play an important role in human infection because in dogs the disease appears long time after infection, and they can move uncontrollably, contributing to disperse the parasite. To take the decision to treat the animals or for euthanasia, in an elimination programme, in order to reduce the parasitic load, it is necessary to diagnose correctly, having more effective tools. Our group has developed a new recombinant antigen-based kinesin-related gene of Leishmania braziliensis (Lbk39), which shows 59% amino acid identity to the L. infantum homologue. The Lbk39 gene was synthesized, inserted into the pLEXSY-sat2 vector and transfected into L. tarentolae cells by electroporation. The recombinant protein was secreted in the culture with a C-terminal histidine marker, purified, generating a product at 337.68 μg mL−1. A total of 152 sera from dog's endemic and non-endemic areas were used, being 78 positives and 75 negatives. The antigen Lbk39 showed 100% sensitivity and 96.1% specificity. We compared this antigen with other antigens such as total extract of the parasite, TRDPP, and our data indicate that Lbk39 has potential application in the diagnosis of CVL through antibody detection.

Epidemic Preparedness—Leishmania tarentolae as an Easy-to-Handle Tool to Produce Antigens for Viral Diagnosis: Application to COVID-19

“…Antigens produced in the L. tarentolae expression system have been assayed for the serological diagnosis of Leishmania infections in humans and dogs, with successful results both in terms of sensibility and sensitivity ( De Souza et al, 2019 ; Rezaei et al, 2019 ; Pereira et al, 2021 ). In addition, in parasites from the Trypanosoma genus, which are phylogenetically related to Leishmania (within the family Trypanosomatidae), antigens produced in L. tarentolae showed satisfactory performances in serological assays, for both African and American trypanosomiases ( Rooney et al, 2015 ; Murphy et al, 2020 ).…”

“…Consequently, L. tarentolae is a very attractive system for antigen production, for both vaccine and diagnostic applications. To date, however, the use of this protozoon for the production of antigens for serodiagnosis has been very limited and has mainly been focused on antigens from protozoans of the genera Leishmania and Trypanosoma , with a sole application in the area of virology, on the hepatitis E virus-HEV ( Baechlein et al, 2013 ; Rooney et al, 2015 ; De Souza et al, 2019 ; Rezaei et al, 2019 ; Murphy et al, 2020 ; Pereira et al, 2021 ).…”

Epidemic preparedness -Leishmania tarentolaeas an easy-to-handle tool to produce antigens for viral diagnosis: application to COVID-19

“…Antigens produced in the L. tarentolae expression system have been assayed for the serological diagnosis of Leishmania infections in humans and dogs, with successful results both in terms of sensibility and sensitivity [30,31,33]. In addition, in parasites from the Trypanosoma genus, which are phylogenetically related to Leishmania (within the family Trypanosomatidae), antigens produced in L. tarentolae showed satisfactory performances in serological assays, for both African and American trypanosomiases [29,32].…”

“…Consequently, L. tarentolae is a very attractive system for antigen production, for both vaccine and diagnostic applications. To date, however, the use of this protozoon for the production of antigens for serodiagnosis has been very limited and has mainly been focused on antigens from protozoans of the genera Leishmania and Trypanosoma, with a sole application in the area of virology, on the hepatitis E virus [28][29][30][31][32][33].…”