Home › Immunology › Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques
Immunology JoVE (Open Access) Citable · DOI

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

DOI: 10.3791/52619-v
What you'll learn
  • ✓ Collect and prepare dried blood spots from skin puncture samples
  • ✓ Properly dry, store, and elute blood spot samples for analysis
  • ✓ Detect viral infection markers using immunoassays on DBS eluates
Protocol

The preparing and processing of dried blood spots (DBS) for their final analysis are still poorly standardized for most diagnostic applications. To overcome this shortcoming, a comprehensive step-by-step protocol is suggested and subsequently evaluated with regard to its effectiveness for detecting markers of viral infections.

Difficulty
intermediate
Total time
~1–2 hours per sample (collection to analysis); storage phase variable (days to weeks)
Biosafety
BSL-2

Steps

1
Collect blood and prepare dried blood spots

Perform skin puncture and apply blood droplets onto filter paper or collection cards. Allow spots to dry naturally at room temperature on a flat, clean surface.

▶ 02:43
2
Dry and store dried blood spot samples

Complete drying process and store prepared blood spots in sealed containers with desiccants at recommended temperature and humidity conditions to preserve sample integrity.

▶ 04:38
3
Elute dried blood spots and analyze eluates

Submerge or incubate dried blood spots in extraction buffer, recover the liquid eluate, and prepare it for immunoassay or molecular detection methods.

▶ 05:40
4
Test eluates for viral infection markers

Perform immunoassays or molecular assays on DBS eluates to detect serological or nucleic acid markers for HBV, HCV, and HIV infections.

▶ 07:39
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