A recombinant cysteine proteinase from Leishmania (Leishmania) chagasi suitable for serodiagnosis of American visceral leishmaniasis.
A recombinant protein, rLdccys1, which was produced by expression of the gene encoding a 30 kDa cysteine proteinase from Leishmania (Leishmania) chagasi, was used for detection of antibodies in sera from patients with active visceral leishmaniasis (VL) in enzyme-linked immunosorbent assays. Analysis of the predicted amino acid sequence of rLdccys1 showed that it contains all the characteristics of a cysteine proteinase. The ability of the protein to react with sera from humans with VL was also shown by Western blotting. The sensitivity for detection of specific antibodies to L. (L.) chagasi bodies using rLdccys1, L. (L.) chagasi promastigote lysates, and amastigote lysates was 80%, 98%, and 99%, respectively. No cross-reactivity between rLdccys1 and Chagas disease was observed, and there was little positive reactivity with sera from patients with cutaneous leishmaniasis and tuberculosis, compared with promastigote and amastigote extracts. Our findings indicate that rLdccys1 from L. (L.) chagasi constitutes a potential tool for the diagnosis of American VL.
de Souza Dias
Suzana
S
Department of Microbiology, Immunology and Parasitology, Universidade Federal de São Paulo, Escola Paulista de Medicina, São Paulo, Brazil. suzi.dias@bol.com.br <suzi.dias@bol.com.br>
da Costa Pinheiro
Paulo Henrique
PH
Katz
Simone
S
dos Santos
Márcia Regina Machado
MR
Barbiéri
Clara Lúcia
CL
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D015153:Blotting, Western; D016022:Case-Control Studies; D006224:Cricetinae; D003546:Cysteine Endopeptidases / Q000235:genetics*; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007891:Leishmania / Q000235:genetics* / Q000276:immunology*; D007898:Leishmaniasis, Visceral / Q000097:blood / Q000175:diagnosis*; D008647:Mesocricetus; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D015800:Protozoan Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D012698:Serologic Tests
D000913:Antibodies, Protozoan; D017931:DNA Primers; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases; C111469:Ldccys1 protein, Leishmania chagasi