Dried blood spot samples: Optimization of commercial EIAs for hepatitis C antibody detection and stability under different storage conditions
| Autor: | B.L.C. Marques, C.U. Brandão, E.F. Silva, V.A. Marques, C.A. Villela-Nogueira, K.M.R. Do Ó, M.T. de Paula, L.L. Lewis-Ximenez, E. Lampe, L.M. Villar |
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| Rok vydání: | 2012 |
| Předmět: |
Adult
Male Coefficient of variation Enzyme-Linked Immunosorbent Assay Sensitivity and Specificity Specimen Handling Young Adult Virology Humans Desiccation Detection limit Reproducibility Chromatography Clinical Laboratory Techniques Chemistry Temperature Hepatitis C antibody Curve analysis Hepatitis C Antibodies Middle Aged Serum samples Hepatitis C Dilution Dried blood spot Blood Infectious Diseases Female |
| Zdroj: | Journal of Medical Virology. 84:1600-1607 |
| ISSN: | 0146-6615 |
| Popis: | This study was undertaken to optimize and compare the efficiency of two commercial EIAs for anti-HCV detection (HCV Ab Radim, Pomezzia, Italy and ETI-AB-HCVK-4 DiaSorin, Vercelli, Italy), in dried blood spot (DBS) samples. The long-term stability of anti-HCV on DBS samples stored at three environmental conditions was also evaluated at: 2-8 °C, 20-25 °C, and -20 °C. Paired DBS and serum samples were obtained from individuals with or without anti-HCV. The type of elution buffer, sample and conjugate volume, sample incubation time and cut-off values were evaluated. For both EIAs, a larger sample volume was used, and the cut-off value determined by the manufacturer was employed for Radim EIA; however, ROC curve analysis was used for the DiaSorin EIA. The sensitivity and specificity of Radim EIA on DBS were 97.5% and 99.5%, respectively, and of DiaSorin EIA were 88.9% and 98.9%, respectively. Accurate results were obtained for a period of 117 days using DBS samples stored at all storage conditions, but storage at -20 °C resulted in the lowest variation among the absorbance values. Both EIAs demonstrated the same limit of detection (until dilution of 1:10(4) with estimated viral load of 3.1 × 10(-1) UI/ml), but the Radim EIA was associated with the best performance because a low coefficient of variation was observed in the repetition and reproducibility studies. In conclusion, commercial EIAs can be optimized for anti-HCV detection in DBS samples that are extremely stable at different conditions for more than 100 days. |
| Databáze: | OpenAIRE |
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