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Updated: Oct 5, 2025
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Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques
Published on: March 13, 2015
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Dried blood spot sampling for hepatitis B virus quantification, sequencing and mutation detection.
Cristianne Sousa Bezerra1,2, Moyra Machado Portilho3, Jakeline Ribeiro Barbosa1
1Laboratorio de Hepatites Virais, Instituto Oswaldo Cruz, Helio and Peggy Pereira Pavillion - Ground Floor - Room B09, Fundação Oswaldo Cruz, FIOCRUZ, Av. Brasil, 4365 - Manguinhos, Rio de Janeiro, Rio de Janeiro, 210360-040, Brazil.
Scientific Reports
|February 1, 2022
Summary
Dried blood spot (DBS) samples offer a viable alternative for Hepatitis B virus (HBV) DNA detection, quantification, and sequencing. This method improves HBV diagnosis accessibility in low-income regions.
Area of Science:
- Virology
- Molecular Diagnostics
- Public Health
Background:
- Hepatitis B virus (HBV) diagnosis typically relies on serum samples, posing accessibility challenges in resource-limited settings.
- Dried blood spot (DBS) sampling offers a simplified approach for sample collection, storage, and transport, bypassing the need for specialized infrastructure.
Purpose of the Study:
- To evaluate the efficacy of DBS samples for the detection, quantification, and sequencing of HBV DNA using in-house molecular techniques.
- To assess the sensitivity and specificity of DBS-based PCR assays compared to traditional serum-based methods.
Main Methods:
- In-house qualitative and quantitative PCR assays were developed and applied to paired serum and DBS samples from 92 HBsAg-positive individuals and 49 controls.
- Sequencing and phylogenetic analyses were performed on HBV DNA extracted from both serum and DBS samples.
- Sensitivity, specificity, and detection limits were determined for DBS-based assays.
Main Results:
- Qualitative PCR on DBS showed 75% sensitivity and 100% specificity (Kappa=0.689) compared to serum.
- Quantitative PCR on DBS had a detection limit of 852.5 copies/mL (250 IU/mL), with 77.63% sensitivity and 100% specificity (Kappa=0.731).
- Sequencing of 63 serum and 36 DBS samples revealed genotypes A, D, E, and F with 100% concordance between sample types; polymorphisms and drug resistance mutations were also identified.
Conclusions:
- DBS samples are a suitable alternative for HBV DNA detection, quantification, and genetic characterization.
- The use of DBS can significantly enhance HBV diagnostic capabilities in low-income regions, contributing to improved HBV control efforts.
