title: Comparative study of amplification systems in immunoenzyme assays for the diagnosis of American tegumentary leishmaniasis.
abstract: We compared the accuracy and reliability of three amplification systems for enzyme immunoassays in the detection of specific IgG antibodies for the diagnosis of cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis in patients from an endemic area in Rio de Janeiro, Brazil. Partially soluble antigens obtained from the promastigote forms of L. (V.) braziliensis were used. For development of the reaction, two chromogens, 1,2-orthophenylenediamine (OPD) and 3,3',5,5'-tetramethylbenzidine (TMB), and a fluorogen, 4-methylumbelliferylphosphate (MUP), were tested. The performance of each system was compared using the following parameters: accuracy, intraclass correlation coefficient (ICC), and area under the receiver operating characteristic (ROC) curve. Sensitivity was the same (97.4%) for all systems. The reliability was excellent (ICC = 98.6, 98.7, and 99.1%) and specificity was 93.7, 95.8, and 97.4% for OPD, MUP, and TMB, respectively, showing no statistical significance. Despite the absence of differences in the performance of the three systems, the use of TMB is suggested because of its operational advantages, such as low cost compared with fluorogens, easy manipulation, greater stability, and lower toxicity.
authors.affiliation: Laboratório de Vigilância em Leishmanioses, Instituto de Pesquisa Clínica Evandro Chagas-IPEC/FIOCRUZ, Rio de Janeiro, RJ, Brazil. lilian.nascimento@ipec.fiocruz.br
authors.forename: Lílian Dias
authors.initials: LD
authors.lastname: Nascimento
authors.forename: Sonia Regina Lambert
authors.initials: SR
authors.lastname: Passos
authors.forename: Eliame
authors.initials: E
authors.lastname: Mouta-Confort
authors.forename: Marta de Almeida
authors.initials: Mde A
authors.lastname: Santiago
authors.forename: Andreia Silva
authors.initials: AS
authors.lastname: Alves
authors.forename: Maria de Fátima
authors.initials: Mde F
authors.lastname: Madeira
authors.forename: Armando de Oliveira
authors.initials: Ade O
authors.lastname: Schubach
authors.forename: Mauro Célio de Almeida
authors.initials: MC
authors.lastname: Marzochi
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology; D001560:Benzidines / Q000737:chemistry; D016001:Confidence Intervals; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives / Q000737:chemistry; D007124:Immunoenzyme Techniques / Q000379:methods*; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D007892:Leishmania braziliensis / Q000276:immunology; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis*; D010655:Phenylenediamines / Q000737:chemistry; D011237:Predictive Value of Tests; D012372:ROC Curve; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000953:Antigens, Protozoan; D001560:Benzidines; D007074:Immunoglobulin G; D010655:Phenylenediamines; C005359:4-methylumbelliferyl phosphate; C021758:3,3',5,5'-tetramethylbenzidine; D006923:Hymecromone; C034193:1,2-diaminobenzene