Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques

Grüner, Nico; Stambouli, Oumaima; Roß, R. Stefan

doi:10.3791/52619

Cited by 125 publications

(115 citation statements)

References 43 publications

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“…The supernatant was immediately analyzed in accordance with manufacturer assay protocols. Dried blood spots were prepared and processed following Clinical & Laboratory Standards Institute guidelines (37). Additional information on sample collection, fabrication, analytical methods, and statistical analysis is provided in Supporting Information.…”

Section: Methodsmentioning

confidence: 99%

Silk-based blood stabilization for diagnostics

Kluge

Kahn

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Advanced personalized medical diagnostics depend on the availability of high-quality biological samples. These are typically biofluids, such as blood, saliva, or urine; and their collection and storage is critical to obtain reliable results. Without proper temperature regulation, protein biomarkers in particular can degrade rapidly in blood samples, an effect that ultimately compromises the quality and reliability of laboratory tests. Here, we present the use of silk fibroin as a solid matrix to encapsulate blood analytes, protecting them from thermally induced damage that could be encountered during nonrefrigerated transportation or freeze-thaw cycles. Blood samples are recovered by simple dissolution of the silk matrix in water. This process is demonstrated to be compatible with a number of immunoassays and provides enhanced sample preservation in comparison with traditional air-drying paper approaches. Additional processing can remediate interactions with conformational structures of the silk protein to further enhance blood stabilization and recovery. This approach can provide expanded utility for remote collection of blood and other biospecimens empowering new modalities of temperature-independent remote diagnostics.lood contains a variety of proteins, enzymes, lipids, metabolites, and peptides, which can be interrogated as biomarkers for health screening, monitoring, and diagnostics. The integrity of these blood components and thus the quality of information attained from their analysis is determined by the storage conditions from sampling until analysis, or the so-called pre-analytical phase (1, 2). This phase often includes time-consuming processing steps and the requirement for a continuous cold storage. Without temperature regulation, blood-derived biospecimens degrade quickly, accounting for up to 67% of all laboratory testing errors (2, 3). Further, when blood-derived materials are frozen, decreases in thermodynamic free energy and unfavorable ice crystal-protein interactions (4) can occur during subsequent thawing (5, 6), which can further compromise analyte integrity as a gold-standard methodology.As an alternative route, blood and blood derivatives can be dried via newer approaches such as isothermal vitrification (7), lyophilization (8), or on silica chips (9). Isothermal vitrification and lyophilization are inherently resource-intensive techniques and thus not suitable for field use. Silica chips are designed for enrichment of selected fractions of the low-molecular-weight serum proteome, but are not broadly protective at elevated temperature (10). An inexpensive alternative used since the 1960s are dried blood spots (DBS) (11), a paper card system which captures blood components among cellulose fibers as the water phase evaporates. These drying measures decrease sample weight by >90%, thus decreasing transport burden, and in theory can enhance long-term sample stability by decreasing water-dependent analyte degradation caused by hydrolysis and enzyme activity (12). Unfortunately, DBS stored in ...

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Section: Methodsmentioning

confidence: 99%

Silk-based blood stabilization for diagnostics

Kluge

Kahn

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…DBS sampling is being increasingly used to facilitate access to serological and NAT for HIV, hepatitis B and C, and other infectious diseases in remote and under resourced regions with poor access to laboratory services, as well as for large epidemiological surveillance studies [171,172].…”

Section: Recommendations For Monitoring Of Treatment Response and Ratmentioning

confidence: 99%

Who to test and how to test for chronic hepatitis C infection – 2016 WHO testing guidance for low- and middle-income countries

Easterbrook

2016

Journal of Hepatology

114

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“…The sample volumes required for DBS are smaller and can remain stable for months until they are processed 6. The processing of DBS samples requires an additional sample extraction step prior to analysis7 making them more costly, with fewer laboratories accredited to analyse them.…”

Section: Introductionmentioning

confidence: 99%