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Comparison of available methods to elute serum from dried blood spot samples for measles serology
Sara Mercader1, David Featherstone, William J Bellini
1Measles, Mumps, Rubella and Herpes Viruses Branch, Division of Viral Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, M.S. C-22, Atlanta, GA 30333, USA. SMercader@cdc.gov
Insights
Comparing six serum extraction methods from dried blood spots, researchers found most effective for measles IgM and IgG detection. A rapid, easy protocol is proposed for outbreak situations.
Area of Science:
- Immunology
- Virology
- Laboratory Medicine
Background:
- Dried blood spots (DBS) are a minimally invasive method for collecting blood samples.
- Accurate serological testing from DBS requires efficient serum extraction protocols.
- Optimizing DBS processing is crucial for infectious disease surveillance, particularly for measles and rubella.
Purpose of the Study:
- To compare the efficacy of six existing serum extraction protocols from filter paper-based dried blood spots.
- To evaluate protocols based on measles IgM and IgG detection, eluate volume, reproducibility, time, cost, and ease of use.
- To identify and propose an optimal protocol for rapid measles/rubella serology in outbreak settings.
Main Methods:
- Six distinct serum extraction protocols were applied to dried blood spots.
- Extracted eluates were assessed for measles IgM and IgG using Dade Behring Enzygnost immunoassays.
- Key performance indicators included detection accuracy, eluate volume, processing time, throughput, reproducibility, and cost-effectiveness.
Main Results:
- Four protocols demonstrated comparable measles IgM detection to serum; two showed significant differences (p < 0.05).
- Measles-specific IgG detection varied significantly between protocols for indeterminate DBS (p < 0.05), but not for positive/negative DBS.
- Sufficient eluate volumes for IgM and IgG assays were obtained from most protocols, with two exceptions for IgM.
Conclusions:
- Most compared protocols effectively extracted antibodies from DBS for serology, with variations in eluate volume, turnaround time, equipment, and cost.
- Two protocols were notably faster (<2 hours processing time).
- An easy-to-implement, rapid serum extraction protocol is proposed for measles/rubella serology in WHO Global Measles and Rubella Laboratory Network outbreak situations.
Abstract:
Six existing protocols for the extraction of serum from blood spots dried onto filter paper were compared. Assessment criteria included: detection of measles IgM and IgG by the Dade Behring Enzygnost immunoassays, volumes of recovered eluates, reproducibility, processing time and throughput, difficulty of protocol, equipment required, safety and estimated costs. Detection of measles IgM in eluates obtained by four of these protocols was as in serum, and significant differences were only observed in eluates from the two remaining protocols (p < 0.05). Significant differences were found between extraction protocols regarding measles-specific IgG detection when an IgG indeterminate DBS was analyzed (p < 0.05), but not when an IgG positive and negative DBS were studied. Sufficient eluate volumes were recovered for testing in the IgM Behring assay following all protocols but two. Sufficient eluate was recovered for testing in the IgG Behring assay following all six protocols. While all protocols were relatively easy to perform, only two protocols required less than 2h for completion. In general, compared protocols performed well on the extraction of antibodies from DBS for serology with differences being observed with eluate volume recovery, turn around time, required equipment and cost. An easy-to-implement protocol is proposed for the rapid extraction of serum for measles/rubella serology in outbreak situations for use in the World Health Organization Global Measles and Rubella Laboratory Network.
