title: Recombinant Leishmania (Leishmania) infantum Ecto-Nucleoside Triphosphate Diphosphohydrolase NTPDase-2 as a new antigen in canine visceral leishmaniasis diagnosis.
abstract: Canine visceral leishmaniasis is an important public health concern. In the epidemiological context of human visceral leishmaniasis, dogs are considered the main reservoir of Leishmania parasites; therefore, dogs must be epidemiologically monitored constantly in endemic areas. Furthermore, dog to human transmission has been correlated with emerging urbanization and increasing rates of leishmaniasis infection worldwide. Leishmania (Leishmania) infantum (L. chagasi) is the etiologic agent of visceral leishmaniasis in the New World. In this work, a new L. (L.) infantum (L. chagasi) recombinant antigen, named ATP diphosphohydrolase (rLic-NTPDase-2), intended for use in the immunodiagnosis of CVL was produced and validated. The extracellular domain of ATP diphosphohydrolase was cloned and expressed in the pET21b-Escherichia coli expression system. Indirect ELISA assays were used to detect the purified rLic-NTPDase-2 antigen using a standard canine sera library. This library contained CVL-positive samples, leishmaniasis-negative samples and samples from Trypanosoma cruzi-infected dogs. The results show a high sensitivity of 100% (95% CI=92.60-100.0%) and a high specificity of 100% (95% CI=86.77-100.0%), with a high degree of confidence (k=1). These findings demonstrate the potential use of this recombinant protein in immune diagnosis of canine leishmaniasis and open the possibility of its application to other diagnostic approaches, such as immunochromatography fast lateral flow assays and human leishmaniasis diagnosis.
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000, MG, Brazil. ronnyfrsouza@yahoo.com.br
authors.forename: Ronny Francisco
authors.initials: RF
authors.lastname: de Souza
authors.forename: Yaro Luciolo
authors.initials: YL
authors.lastname: Dos Santos
authors.forename: Raphael
authors.initials: R
authors.lastname: de Souza Vasconcellos
authors.forename: Lucas
authors.initials: L
authors.lastname: Borges-Pereira
authors.forename: Ivo Santana
authors.initials: IS
authors.lastname: Caldas
authors.forename: Márcia Rogéria
authors.initials: MR
authors.lastname: de Almeida
authors.forename: Maria Terezinha
authors.initials: MT
authors.lastname: Bahia
authors.forename: Juliana Lopes Rangel
authors.initials: JL
authors.lastname: Fietto
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics; D019411:Clinical Laboratory Techniques / Q000379:methods*; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004283:Dog Diseases / Q000175:diagnosis* / Q000469:parasitology; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D018314:Leishmania infantum / Q000302:isolation & purification*; D007898:Leishmaniasis, Visceral / Q000175:diagnosis / Q000662:veterinary*; D008969:Molecular Sequence Data; D010274:Parasitology / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D011994:Recombinant Proteins; D000251:Adenosine Triphosphatases; C027201:ectoATPase