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Please use this identifier to cite or link to this item: http://hdl.handle.net/1807/19580

Title: Validation of an ELISA for the Quantitation of Diphtheria Antitoxin in Human Serum
Issue Date: 31-Dec-2000
Publisher: Elfos Scientiae
Citation: Biotecnologia Aplicada (ISSN: 0684-4551) Vol 17 Num 3
Abstract: Biotecnologa Aplicada, Volume 17, July-September 2000, pp. 183-186 Validation of an ELISA for the Quantitation of Diphtheria Antitoxin in Human Serum Code Number: BA00053 ABSTRACT: An indirect solid-phase enzyme-linked immunosorbent assay (ELISA) was developed for the quantitation of antidiphtheria antibodies in human sera on the basis of a calibrated antibody standard. Intra- and inter-assay imprecision was around 10%. This assay showed an excellent accuracy (97.06% average recovery). Parallelism deviations were below 10%, which were evaluated with dilutions covering the working range of the standard curve. The detection limit of 0.0044 international units per milliliter (IU/mL) was sufficient for determining the immunological protection against diphtheria. In a preliminary study, a good correlation was found between the ELISA and the neutralization test in guinea pigs (R2 = 0.992). A wide range of diphtheria antitoxin quantitation made possible to classify 81 adults prior and four weeks after revaccination against diphtheria as: unprotected subjects ( 0.01 U/mL), relatively protected (> 0.010.1 IU/mL), satisfactorily protected (> 0.11.0 IU/mL), and long-lasting protected (>1.0 IU/mL). Keywords: diphtheria antitoxin, ELISA, validation RESUMEN: Validacin de un ELISA para la cuantificacin de anticuerpos antitoxina diftrica en sueros humanos. Se desarroll un ensayo inmunoenzimtico en fase slida (ELISA) de tipo indirecto para la cuantificacin de antitoxina diftrica en sueros humanos, con el empleo de un estndar previamente calibrado. Las imprecisiones intra e interensayo fueron de alrededor de 10%. Este ensayo tambin mostr una excelente precisin (recobrado medio de 97,06%). Las desviaciones del paralelismo estuvieron por debajo de 10%, las cuales se evaluaron con el empleo de diluciones que cubran el intervalo de trabajo de la curva estndar. El lmite de deteccin de 0,0044 unidades internacionales por mililitro (UI/mL) fue suficiente para determinar el grado de proteccin inmunolgica contra la difteria. En un estudio preliminar, el ensayo mostr una correlacin adecuada con la prueba de neutralizacin en conejillos de Indias (R2 = 0,992). Este inmunoensayo ofrece, adems, un amplio intervalo de cuantificacin, lo que permiti la clasificacin de 81 individuos adultos antes y cuatro semanas despus de la revacunacin como: desprotegidos ( 0,01 UI/mL), relativamente protegidos (> 0,01-0,1 UI/mL), satisfactoriamente protegidos (> 0,1-1,0 UI/mL) y protegidos por largo tiempo (> 1,0 UI/mL). Palabras claves: antitoxina diftrica, ELISA, validacin Copyright 2000 Elfos Scientiae
URI: http://hdl.handle.net/1807/19580
Other Identifiers: http://www.bioline.org.br/abstract?id=ba00053
Rights: Copyright 2000 Elfos Scientiae
Appears in Collections:Bioline International Legacy Collection

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