A strategy for identifying serodiagnostically relevant antigens of Leishmania or other pathogens in genetic libraries.
Different individuals, when infected with the same parasite, rarely produce antibodies against the same set of antigens. Indeed, unless a particular antigen happens to be recognized by antibodies in all individuals, the use of a single antigen in the serodiagnosis of parasitic diseases leads, invariably, to false-negative results. A straightforward method for pin-pointing, in genetic libraries, the precise antigens that would increase serodiagnostic assay sensitivities is presented. The method is based on the utilization of sera that produced false-negative results against previously available antigens. Employing this false-negative serum-selection methodology for the identification of new Leishmania infantum recombinant antigens (rAgs), the sensitivity of a dipstick assay for anti-Leishmania antibodies in a panel of sera from patients with visceral leishmaniasis was increased from 83.3% to 98.1%, without affecting its specificity, by the inclusion of a fifth and a sixth L. infantum rAg.
Centro de Pesquisas Gonçalo Moniz, Fundação Oswaldo Cruz, Rua Valdemar Falcão 121, 40295-001, Salvador, Brazil.
Márcia C A
MC
Teixeira
Geraldo G S
GG
Oliveira
Marco A
MA
Silvany
Neuza M
NM
Alcântara-Neves
Milena B P
MB
Soares
Ricardo
R
Ribeiro-Dos-Santos
Selma M B
SM
Jerônimo
Carlos H
CH
Costa
Washington L C
WL
dos-Santos
Daniel
D
Eichinger
Lain
L
Pontes-de-Carvalho
D000818:Animals; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan / Q000302:isolation & purification*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005188:False Negative Reactions; D015723:Gene Library; D006801:Humans; D007891:Leishmania / Q000302:isolation & purification*; D007898:Leishmaniasis, Visceral / Q000097:blood; D010271:Parasites / Q000302:isolation & purification*; D012016:Reference Values; D012680:Sensitivity and Specificity
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan