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Centro de Investigaciones Regionales Dr. Hideyo Noguchi, Universidad Autónoma de Yucatán

2004, Number 1

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Rev Biomed 2004; 15 (1)

Response of IgM antibodies against hepatitis A virus immunogenic epitopes

Delahanty-Fernández A, Valdivia-Alvarez I, Trujillo-Brito J, Hernández-Marin M, Gómez-Cordero I, Ventura-Paz J, Palenzuela-Díaz A, Acosta-Bas C, Zulueta-Rodríguez O, Rodríguez AM

Language: Spanish
References: 19
Page: 11-16
PDF size: 39.63 Kb.


ABSTRACT

Introduction. Hepatitis A is a disease characterized by acute inflammation of the liver, as a consequence of infection by hepatitis A virus (HAV). Due to the difficulties in obtaining the natural antigen of the HAV using cell culture, peptide synthesis technology has been used for the simulation of the main antigenic sites of the virus. The objective of this work was to determine the IgM antibody response against three immunogenic epitopes from the structural regions VP1 and VP3 of HAV.
Materials and Methods. The peptides were synthesized in solid phase, following Merrifield´s method. The purity was determined with the High Performance Liquid Chromatography system by means of reverse chromatography. 38 serum samples of HAV positive patients and 90 negative samples of blood donors were evaluated in an UMELISA assay. The MicroElisa HAV IgM (Organon Teknika) was used as a confirmatory assay.
Results. Two peptides of the VP1 region were synthesized (b4 and b6) and one peptide of the VP3 region (b9), and a main signal was obtained in all the chromatograms. For peptide b4, 81.82% of samples had a fluorescence/cutoff relation between 2 and 5, 18.18% over 10; for b6 and b9 ratios were between 1 and 5. Reactivity figures were low, moderate, and high. The most recognizable peptide was b4 from region VP1.
Conclusions. These results show that assays using syntetic peptides can be an apropiate tool in the development of ELISA-type techniques for the detection of infection by HVA.


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