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Background  Transfusion‐transmitted babesiosis caused by   B abesia microti  has emerged as a significant risk to the  US  blood supply. This study estimated the prevalence of   B . microti  antibodies in blood donors using an investigational enzyme immunoassay ( EIA ).    Study Design and Methods  A peptide‐based  EIA  that detects both immunoglobulin ( Ig ) G  and  IgM  antibodies to   B . microti  was developed and validated. Donor samples randomly selected from areas defined as high‐risk endemic, lower‐risk endemic, and nonendemic for   B . microti  were deidentified and tested using the investigational  EIA . Samples that were  EIA  repeat reactive were further tested by   B . microti  immunofluorescent assay ( IFA ), polymerase chain reaction ( PCR ) on red blood cell lysates, and peripheral blood smear examination. A random subset of 1272 samples from high‐risk endemic areas was tested by  IFA ,  PCR , and peripheral blood smear in parallel with  EIA .    Results  Among 15,000 donations tested with the investigational   B . microti   EIA ,  EIA  repeat‐reactive rates were 1.08% (54/5000) in a high‐risk endemic area, 0.74% (37/5000) in a lower‐risk area, and 0.40% (20/5000) in a nonendemic area. After application of a revised cutoff, these values were reduced to 0.92%, (46/5000), 0.54% (27/5000), and 0.16% (8/5000). Overall concordance between  EIA  and  IFA  among donor samples was 99.34%. One seropositive sample was positive by  PCR .    Conclusion  The seroprevalence of   B . microti  in blood donors in a high‐risk area measured by an investigational  EIA  was approximately 1%. The  EIA  shows promise as an efficient high‐throughput blood donor screening assay for   B . microti .

Determination of B abesia microti seroprevalence in blood donor populations using an investigational enzyme immunoassay