Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology · 2025
Long-term stability of dried blood spot samples for HIV diagnosis in adults from Kinshasa (Democratic Republic of Congo).
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Astiz Nerea, Martínez de Aguirre Paula, Carlos Silvia, Mbikayi Samclide, Barquín David, Burgueño Eduardo, Holguín África, Reina Gabriel
Аннотация
BACKGROUND: Dried blood spots (DBS) are a widely used sampling method for serological testing, particularly in resource-limited settings. However, long-term stability of HIV antibodies in DBS remains underexplored. This study evaluates the stability of HIV serological markers stored at -80 °C for 7 years and compares the analytical performance and concordance of three different diagnostic techniques: electrochemiluminescence immunoassay (ECLIA), enzyme-linked fluorescent assay (ELFA), and immunochromatography.
METHODS: Between 2016 and 2017, a total of 143 DBS samples were collected at Monkole Hospital in Kinshasa, D.R. Congo. For each patient, two DBS cards were prepared, each containing five spots of whole blood. The first card was analyzed in 2017 using three serological assays for HIV diagnosis: ECLIA (Roche), ELFA (bioMerieux), and immunochromatography (Geenius, BioRad). The second card was stored at -80 °C until 2024, when it was tested again using the same diagnostic tools.
RESULTS: After 7 years of storage, HIV markers in DBS samples showed excellent stability. Sensitivity and specificity were 100 % for all techniques, except ECLIA, which produced one false positive. Inter-assay agreement was high in both 2017 and 2024, with Kappa values ranging from 0.90 to 1.00. Spearman correlation coefficients between 2017 and 2024 results were strong: 0.99 for ECLIA, 0.89 for ELFA, and 0.77-0.81 for immunochromatography. A significant increase in test values were observed for ELFA in 2024 but diagnostic classification remained unchanged.
CONCLUSIONS: High sensitivity, specificity, and inter-assay concordance support the long-term use of DBS for HIV diagnosis and surveillance. These findings can be particularly relevant for biobanking and retrospective seroepidemiology.
METHODS: Between 2016 and 2017, a total of 143 DBS samples were collected at Monkole Hospital in Kinshasa, D.R. Congo. For each patient, two DBS cards were prepared, each containing five spots of whole blood. The first card was analyzed in 2017 using three serological assays for HIV diagnosis: ECLIA (Roche), ELFA (bioMerieux), and immunochromatography (Geenius, BioRad). The second card was stored at -80 °C until 2024, when it was tested again using the same diagnostic tools.
RESULTS: After 7 years of storage, HIV markers in DBS samples showed excellent stability. Sensitivity and specificity were 100 % for all techniques, except ECLIA, which produced one false positive. Inter-assay agreement was high in both 2017 and 2024, with Kappa values ranging from 0.90 to 1.00. Spearman correlation coefficients between 2017 and 2024 results were strong: 0.99 for ECLIA, 0.89 for ELFA, and 0.77-0.81 for immunochromatography. A significant increase in test values were observed for ELFA in 2024 but diagnostic classification remained unchanged.
CONCLUSIONS: High sensitivity, specificity, and inter-assay concordance support the long-term use of DBS for HIV diagnosis and surveillance. These findings can be particularly relevant for biobanking and retrospective seroepidemiology.