Synthetic peptide-based enzyme-linked immunosorbent assay for serodiagnosis of visceral leishmaniasis.
Synthetic peptides, derived from the amino acid sequence of a Leishmania donovani clone, were used to develop an enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against L. donovani. For this purpose, five peptides were conjugated to a protein carrier, human serum albumin (HSA), by using a heterobifunctional reagent, epsilon-maleimidocaproic acid N-hydroxysuccinimide ester, to obtain a well-defined product. The sensitivity and the specificity of the peptide-specific ELISA were determined with a panel of 106 serum samples from individuals living in areas where visceral leishmaniasis is endemic; sera from post-kala azar dermal leishmaniasis-infected patients and from individuals suffering from other infectious diseases were also included. ELISAs were performed with either a single peptide-HSA conjugate or a mixture of two peptide-HSA conjugates. Ninety-seven percent of the serum samples from patients with visceral leishmaniasis had detectable antibodies to one or more of the single synthetic peptides. ELISA with a single peptide-HSA conjugate proved to be less sensitive (less than 71%) but more specific (up to 93%) than ELISA with crude promastigote antigens (80% sensitivity and 79% specificity); when a combination of two different peptide-HSA conjugates was used, the test increased both in sensitivity and in specificity. Chemically defined peptide-protein conjugates improve the reproducibility and reliability of ELISA for the serodiagnosis of L. donovani infection.
Fargeas
C
C
Glycobiologie, Centre National de la Recherche Scientifique, Orléans, France.
Hommel
M
M
Maingon
R
R
Dourado
C
C
Monsigny
M
M
Mayer
R
R
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000918:Antibody Specificity; D000953:Antigens, Protozoan / Q000235:genetics; D001483:Base Sequence; D003432:Cross-Linking Reagents; D016054:DNA, Protozoan / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D005069:Evaluation Studies as Topic; D006801:Humans; D007893:Leishmania donovani / Q000235:genetics / Q000276:immunology; D007898:Leishmaniasis, Visceral / Q000175:diagnosis* / Q000276:immunology; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000235:genetics / Q000276:immunology; D010455:Peptides / Q000138:chemical synthesis / Q000235:genetics / Q000276:immunology; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods* / Q000706:statistics & numerical data; D012709:Serum Albumin
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D003432:Cross-Linking Reagents; D016054:DNA, Protozoan; D009842:Oligopeptides; D010455:Peptides; D015800:Protozoan Proteins; D012709:Serum Albumin; C047981:arginyl-glycyl-aspartic acid