Comparison of available methods to elute serum from dried blood spot samples for measles serology.
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.
Mercader
Sara
S
Measles, 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
Featherstone
David
D
Bellini
William J
WJ
D000914:Antibodies, Viral / Q000097:blood*; D001800:Blood Specimen Collection; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D008457:Measles / Q000175:diagnosis / Q000276:immunology*; D008459:Measles virus / Q000276:immunology*; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*; D044967:Serum / Q000276:immunology*
D000914:Antibodies, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011933:Reagent Kits, Diagnostic