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Updated: Jul 31, 2026
Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques
Published on: March 13, 2015
Optimization and Stability Testing of Four Commercially Available Dried Blood Spot Devices for Estimating Measles and
Ojas Kaduskar1, Vaishali Bhatt1, Christine Prosperi2
1Diagnostic Virology Group, ICMR-National Institute of Virology, Pune, Maharashtra, India.
Insights
Dried blood spot (DBS) collection is a convenient alternative to venipuncture for measles and rubella IgG antibody testing. Optimized protocols ensure accurate results comparable to serum, with good stability at ambient temperatures.
Area of Science:
- Immunology
- Virology
- Public Health
Background:
- Dried blood spot (DBS) collection offers advantages for sample collection, transport, and storage over traditional venipuncture.
- However, DBS requires additional processing, potentially introducing variability in results.
Purpose of the Study:
- To optimize elution protocols for DBS devices for accurate measurement of antimeasles and antirubella IgG antibodies.
- To assess the stability of DBS samples under various storage and transport conditions.
Main Methods:
- Whole-blood samples were collected on four DBS devices and compared with paired serum samples.
- Elution protocols were optimized, and antibody concentrations were measured using enzyme immunoassay.
- Stability of HemaSpot HF DBS devices was evaluated at different temperatures and time points, including field conditions.
Main Results:
- Three DBS devices showed comparable antibody concentrations to serum after protocol optimization.
- HemaSpot HF DBS devices maintained antibody stability for 90 days at 4°C and 30 days at ambient temperature (22-25°C).
- Elevated temperatures (45°C) caused significant antibody decline, while field transport resulted in quantitative, but not qualitative, variability.
Conclusions:
- DBS devices, with optimized elution, provide an accurate alternative to serum for measuring measles and rubella IgG antibodies.
- The stability of HemaSpot HF at ambient temperatures supports its use in serosurveys where cold storage is not feasible.
Abstract:
Blood collection using dried blood spots (DBS) provides an easier alternative to venipuncture for sample collection, transport, and storage but requires additional processing that can cause variability in results. Whole-blood samples spotted on four DBS devices and respective paired serum samples were tested for antimeasles and antirubella IgG antibody concentrations by enzyme immunoassay. Elution protocols for DBS devices were optimized for comparability relative to serum samples using 12 adult volunteers. Stability of DBS collected on HemaSpot HF was assessed under various temperature conditions (+4, 22 to 25, and 45°C) at six time points (0, 7, 15, 30, 60, and 90 days) in a controlled laboratory setting using six adult volunteers. Devices were shipped and stored for 30 days at four settings with variable temperature and humidity conditions to assess the impact on antibody concentrations. Three DBS devices demonstrated comparable antibody concentrations with paired sera following optimization. Antibodies recovered from DBS were stable for at least 90 days at 4°C and for 30 days at ambient temperature (22 to 25°C) using the HemaSpot HF device. A drastic decline in antibody concentrations was observed at 45°C, resulting in quantitative and qualitative discrepancies by day 7. HemaSpot HF devices shipped to field sites and stored at ambient temperature and humidity resulted in quantitative, but not qualitative, variability. Measurement of antimeasles and antirubella IgG antibodies with DBS devices is an accurate alternative to testing serum, provided elution protocols are optimized. Stability of HemaSpot HF devices at ambient temperature enables broader use in surveys when serum processing and cold storage are not feasible. IMPORTANCE Dried blood spot (DBS) collection offers various advantages over conventional methods of blood collection, especially when collecting and transporting samples for a serosurvey. Yet use of DBS requires additional processing steps in the laboratory that can add to variability in results. We optimized a protocol to elute IgG antibodies against measles and rubella viruses in four DBS devices, demonstrating high concordance with paired venous sera for most devices. Extensive stability studies with various temperature and storage conditions in the laboratory and in the field were conducted using HemaSpot HF DBS devices prior to its use in one of the largest community-based measles and rubella serological surveys in the world.
