Article Category:
Research Article

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

Sarah Gwyn Centers for Disease Control and Prevention, Atlanta, Georgia;

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Solomon Aragie The Carter Center Ethiopia, Addis Ababa, Ethiopia;

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Dionna M. Wittberg Francis I. Proctor Foundation, University of California, San Francisco, California;

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Jason S. Melo Francis I. Proctor Foundation, University of California, San Francisco, California;

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Adane Dagnew The Carter Center Ethiopia, Addis Ababa, Ethiopia;

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Dagnachew Hailu The Carter Center Ethiopia, Addis Ababa, Ethiopia;

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Zerihun Tadesse The Carter Center Ethiopia, Addis Ababa, Ethiopia;

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Mahteme Haile Amhara Public Health Institute, Bahir Dar, Ethiopia;

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Taye Zeru Amhara Public Health Institute, Bahir Dar, Ethiopia;

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Scott D. Nash The Carter Center, Atlanta, Georgia;

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Benjamin F. Arnold Francis I. Proctor Foundation, University of California, San Francisco, California;

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Diana L. Martin Centers for Disease Control and Prevention, Atlanta, Georgia;

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Jeremy D. Keenan Francis I. Proctor Foundation, University of California, San Francisco, California;
Department of Ophthalmology, University of California, San Francisco, California

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DOI:
https://doi.org/10.4269/ajtmh.21-0140
Volume/Issue:
Volume 105: Issue 3
Page(s):
822–827
Restricted access
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ABSTRACT.

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.

    • Supplemental Materials (DOCX 1480 KB)

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.

Copyright:
© The American Society of Tropical Medicine and Hygiene 2021
Received:
04 Feb 2021
|
Accepted:
16 Apr 2021
|
Published Online:
12 Jul 2021
Cover The American Journal of Tropical Medicine and Hygiene
The American Journal of Tropical Medicine and Hygiene
Print ISSN:
0002-9637
Online ISSN:
1476-1645
  • 1.

    Arnold BF , Scobie HM , Priest JW , Lammie PJ , 2018. Integrated serologic surveillance of population immunity and disease transmission. Emerg Infect Dis 24: 1188–1194.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 2.

    Gwyn S , Cooley G , Goodhew B , Kohlhoff S , Banniettis N , Wiegand R , Martin DL , 2017. Comparison of platforms for testing antibody responses against the Chlamydia trachomatis antigen Pgp3. Am J Trop Med Hyg 97: 1662–1668.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 3.

    Kaur H , Dize L , Munoz B , Gaydos C , West SK , 2018. Evaluation of the reproducibility of a serological test for antibodies to Chlamydia trachomatis pgp3: a potential surveillance tool for trachoma programs. J Microbiol Methods 147: 56–58.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 4.

    Wittberg DM et al., 2021. WASH Upgrades for Health in Amhara (WUHA): study protocol for a cluster-randomised trial in Ethiopia. BMJ Open 11: e039529.

  • 5.

    Aiemjoy K , Aragie S , Wittberg DM , Tadesse Z , Callahan EK , Gwyn S , Martin D , Keenan JD , Arnold BF , 2020. Seroprevalence of antibodies against Chlamydia trachomatis and enteropathogens and distance to the nearest water source among young children in the Amhara region of Ethiopia. PLoS Negl Trop Dis 14: e0008647.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 6.

    Goodhew EB et al., 2012. CT694 and pgp3 as serological tools for monitoring trachoma programs. PLoS Negl Trop Dis 6: e1873.

  • 7.

    Migchelsen SJ et al., 2017. Defining seropositivity thresholds for use in trachoma elimination studies. PLoS Negl Trop Dis 11: e0005230.

  • 8.

    Priest JW , Moss DM , 2020. Measuring cryptosporidium serologic responses by multiplex bead assay. Methods Mol Biol 2052: 61–85.

  • 9.

    Shrout PE , Fleiss JL , 1979. Intraclass correlations: uses in assessing rater reliability. Psychol Bull 86: 420–428.

  • 10.

    Menditto A , Patriarca M , Magnusson B , 2007. Understanding the meaning of accuracy, trueness and precision. Accredit Qual Assur 12: 45–47.

  • 11.

    Moss DM , Montgomery JM , Newland SV , Priest JW , Lammie PJ , 2004. Detection of cryptosporidium antibodies in sera and oral fluids using multiplex bead assay. J Parasitol 90: 397–404.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12.

    Houpt E , Barroso L , Lockhart L , Wright R , Cramer C , Lyerly D , Petri WA , 2004. Prevention of intestinal amebiasis by vaccination with the Entamoeba histolytica Gal/GalNac lectin. Vaccine 22: 611–617.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13.

    Schmidt-Ott R , Brass F , Scholz C , Werner C , Gross U , 2005. Improved serodiagnosis of Campylobacter jejuni infections using recombinant antigens. J Med Microbiol 54: 761–767.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14.

    Priest JW , Moss DM , Visvesvara GS , Jones CC , Li A , Isaac-Renton JL , 2010. Multiplex assay detection of immunoglobulin G antibodies that recognize Giardia intestinalis and Cryptosporidium parvum antigens. Clin Vaccine Immunol 17: 1695–1707.

    • PubMed
    • Search Google Scholar
    • Export Citation
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