Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples

dc.creatorBezerra, Cristiane Sousa
dc.creatorPortilho, Moyra Machado
dc.creatorFrota, Cristiane Cunha
dc.creatorVillar, Lívia Melo
dc.date2021-06-25T20:02:41Z
dc.date2021-06-25T20:02:41Z
dc.date2021
dc.date.accessioned2023-09-26T21:24:00Z
dc.date.available2023-09-26T21:24:00Z
dc.identifierBEZERRA, Cristiane Sousa et al. Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples. Microbiology Open, v. 10, e1161, 8 p, 2021.
dc.identifier2045-8827
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/47877
dc.identifier10.1002/mbo3.1161
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8872269
dc.descriptionThe dried blood spot (DBS) samples are a useful resource for viral DNA isolation and important in increasing access to HBV diagnosis. However, the choice of the DNA extraction method is crucial for reliable results. We compared the reliability of four DNA extraction methods using DBS samples for the qualitative and quantitative de tection of HBV. A panel of serially diluted HBV DNA in whole blood was spotted onto filter paper (Whatman 903 paper and Whatman FTA cards). Four methods were used to extract DNA: QIAamp® DNA Blood Mini Kit (Qiagen); High Pure Viral Nucleic Acid Kit (Roche); Invisorb Spin Blood Midi Kit (Invitek), and DBS Genomic DNA Isolation Kit (Norgen Biotek). Two qualitative PCRs for the core and surface gene regions of HBV were used, and in-house real-time PCR was also evaluated. It was possible to detect HBV DNA using all extraction and PCR protocols. The lowest limit of detection was found using Whatman 903 paper, Roche extraction, and qualitative PCR (20 copies of HBV DNA per ml) for the surface/polymerase HBV region. In the case of in-house real-time PCR, the lowest limit of detection was found using both Roche and Qiagen assays (estimated 2 × 103 copies per ml). These results suggest the importance of both the extraction method and PCR protocol in detecting HBV DNA in DBS. This study provides insights into the utility of DBS samples in HBV molecular diagnosis and their feasibility in low resource areas where cold storage and transportation may be difficult.
dc.formatapplication/pdf
dc.languageeng
dc.publisherWiley
dc.rightsopen access
dc.subjectDBS
dc.subjectextração de DNA
dc.subjectHepatite B
dc.subjectPCR em tempo real
dc.subjectDBS
dc.subjectDNA extraction
dc.subjectHepatite B
dc.subjectreal-time PCR
dc.titleComparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples
dc.typeArticle


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