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

2017, Number 3

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Rev Biomed 2017; 28 (3)

Validation of an ELISA for quantitation of serum levels of surface antigen in patients with chronic hepatitis B

Rodríguez-Pelier CV, Chang-Monteagudo A, Zúñiga-Rosales Y, Torres-Rives B, Martínez-Tellez G, la Rosa-Hernández D, Martínez-Perera A

Language: Spanish
References: 42
Page: 104-121
PDF size: 379.48 Kb.


ABSTRACT

Introduction. During the acute phase of infection with Hepatitis B Virus (HBV) and the initial period of chronic infection, DNA is episomally (free or extrachromosomal) and replicates in hepatocytes producing among other virions, DNA polymerase and virus surface antigen (HBsAg).
Objective. Validate an ELISA to quantify HBsAg levels in patients with chronic hepatitis B.
Method. An experimental study of technological developmentwas conducted. The normalization and validation of heterogeneous double antibody enzyme linked inmunoassay for quantitation of HBsAg in human sera was carried out. 115 serum samples from patients with chronic hepatitis B with viral load results were correlated with its HBsAg concentrations.
Results and discussion. The developed method presented variation coefficients intra and interassay of 9.8 and 13.2% respectively. The working range was estimated between 0.15 and 60 ng/mL. The recovery rate was between 90 and 110% and the linear fit of the standard curve presented a determination coefficient greater than 0, 99. The correlation between the DNA levels and HBsAg concentration was 62.5%.
Conclusions. The assessment of the ELISA for quantitation of HBsAg developed in the laboratory showed that it achieved validation parameters for its clinical use.


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