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Precision of Serologic Testing from Dried Blood Spots Using a Multiplex Bead Assay

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  • 1 Centers for Disease Control and Prevention, Atlanta, Georgia;
  • | 2 The Carter Center Ethiopia, Addis Ababa, Ethiopia;
  • | 3 Francis I. Proctor Foundation, University of California, San Francisco, California;
  • | 4 Amhara Public Health Institute, Bahir Dar, Ethiopia;
  • | 5 The Carter Center, Atlanta, Georgia;
  • | 6 Department of Ophthalmology, University of California, San Francisco, California
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Multiplex bead assays (MBAs) for serologic testing have become more prevalent in public health surveys, but few studies have assessed their test performance. As part of a trachoma study conducted in a rural part of Ethiopia in 2016, dried blood spots (DBS) were collected from a random sample of 393 children aged 0 to 9 years, with at least two separate 6-mm DBS collected on a filter card. Samples eluted from DBS were processed using an MBA on the Luminex platform for antibodies against 13 antigens of nine infectious organisms: Chlamydia trachomatis, Vibrio cholera, enterotoxigenic Escherichia coli, Cryptosporidium parvum, Entamoeba histolytica, Camplyobacter jejuni, Salmonella typhimurium Group B, Salmonella enteritidis Group D, and Giardia lamblia. Two separate DBS from each child were processed. The first DBS was run a single time, with the MBA set to read 100 beads per well. The second DBS was run twice, first at 100 beads per well and then at 50 beads per well. Results were expressed as the median fluorescence intensity minus background (MFI–BG), and classified as seropositive or seronegative according to external standards. Agreement between the three runs was high, with intraclass correlation coefficients of > 0.85 for the two Salmonella antibody responses and > 0.95 for the other 11 antibody responses. Agreement was also high for the dichotomous seropositivity indicators, with Cohen’s kappa statistics exceeding 0.87 for each antibody assay. These results suggest that serologic testing on the Luminex platform had strong test performance characteristics for analyzing antibodies using DBS.

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Author Notes

Address correspondence to Jeremy D. Keenan, Francis I Proctor Foundation, University of California, San Francisco, 490 Illinois Street, Floor 2, Box 0944, San Francisco, CA 94158. E-mail: jeremy.keenan@ucsf.edu

Financial support: This work was supported by grant U10EY023939 from the National Institutes of Health—National Eye Institute (Bethesda, MD); grant NTDSC 062 from the Coalition for Operational Research on Neglected Tropical Diseases (COR-NTD, funded at The Task Force for Global Health primarily by the Bill & Melinda Gates Foundation, the UK Department for International Development, and the U.S. Agency for International Development through Its Neglected Tropical Diseases Program; Atlanta, GA), That Man May See, and Research to Prevent Blindness.

Disclaimer: The findings presented in this study are those of the authors alone and do not represent the views of the Centers for Disease Control and Prevention.

Authors’ addresses: Sarah Gwyn and Diana L. Martin, Centers for Disease Control and Prevention, Atlanta, GA, E-mails: yme8@cdc.gov and hzx3@cdc.gov. Solomon Aragie, The Carter Center Ethiopia, Addis Ababa, Ethiopia, E-mail: solomon.aragie@cartercenter.org. Dionna M. Wittberg, Jason S. Melo, Benjamin F. Arnold, and Jeremy D. Keenan, Francis I. Proctor Foundation, University of California, San Francisco, San Francisco, CA, E-mails: dionna.wittberg@ucsf.edu, jason.s.melo@gmail.com, ben.arnold@ucsf.edu, and jeremy.keenan@ucsf.edu. Adane Dagnew, Dagnachew Hailu, Zerihun Tadesse, and Scott D. Nash, The Carter Center Ethiopia, Atlanta, GA, E-mails: adane.dagnew@cartercenter.org, dagnachew.hailu@cartercenter.org, zerihun.tadesse@cartercenter.org, and scott.nash@cartercenter.org. Mahteme Haile and Taye Zeru, Amhara Public Health Institute, Bahir Dar, Ethiopia, E-mails: maykm24@yahoo.com and zerutaye@gmail.com.