Interlaboratory comparison of a multiplex bead assay against the Chlamydia trachomatis antigen Pgp3 using two monoclonal antibody clones
5 affiliations
- 1Division of Parasitic Diseases and Malaria, National Center for Zoonotic, Waterborne, and Environmental Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America
- 2Department of Immunology, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom
- 3Infectious Disease Epidemiology and Analytics Unit, Department of Global Health, Institut Pasteur, Université Paris-Cité, Paris, France
- 4Synergy America, Inc., Atlanta, Georgia, United States of America
- 5Eastern and Southern Africa Centre of International Parasite Control, Kenya Medical Research Institute, Nairobi, Kenya
Abstract
The bacterium Chlamydia trachomatis causes three distinct conditions – urogenital chlamydia, the eye disease trachoma, and lymphogranuloma venereum. Use of serological testing for exposure to C. trachomatis for evaluating ocular disease risk factors and population-based surveillance has expanded over the past decade. We developed chimeric monoclonal antibodies to the C. trachomatis antigen Pgp3 as standardized reagents for Pgp3 serological assays. Here we use two anti-Pgp3 monoclonals (1G1 and 1C3) to conduct an interlaboratory comparison of Pgp3 Luminex-based multiplex bead serological assays in 4 different laboratories (each with their own protocol) with 5 operators using 3 different instrument types. Antibodies spotted onto filter paper were stable for two weeks at room temperature. Accuracy and precision were evaluated at 3 concentrations for each monoclonal (high, medium, and low). Accuracy within 20% was achieved for 11/21 conditions for 1G1 and 18/21 conditions for 1C3; the high concentration resulted in the poorest accuracy. Precision was within 10% for all low and medium concentrations and was above 10% for several conditions at the high concentration. The fluorescence values differed between instruments but showed no difference when extrapolated to concentrations using a standard curve. These data suggest that 1C3 may be the more reliable standard reagent and that the Pgp3 multiplex assays are highly robust when comparing different instruments and assay protocols.
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BibTeX
@article{gwyn2026interlaboratory,
title = {{Interlaboratory comparison of a multiplex bead assay against the Chlamydia trachomatis antigen Pgp3 using two monoclonal antibody clones}},
author = {Sarah Gwyn and Karana Wickens and Helena Brazal Monzo and Chris Drakeley and Stephane Pelleau and Michael White and Zachary Reynolds and Henry Kanyi and Richelle W. Kihoro and Henry Emisiko and Sammy M. Njenga and Diana L. Martin},
journal = {PLOS Global Public Health},
year = {2026},
month = nov,
doi = {10.1371/journal.pgph.0007564},
url = {https://journals.plos.org/globalpublichealth/article?id=10.1371/journal.pgph.0007564},
note = {CC0},
}
RIS
TY - JOUR
TI - Interlaboratory comparison of a multiplex bead assay against the Chlamydia trachomatis antigen Pgp3 using two monoclonal antibody clones
AU - Sarah Gwyn
AU - Karana Wickens
AU - Helena Brazal Monzo
AU - Chris Drakeley
AU - Stephane Pelleau
AU - Michael White
AU - Zachary Reynolds
AU - Henry Kanyi
AU - Richelle W. Kihoro
AU - Henry Emisiko
AU - Sammy M. Njenga
AU - Diana L. Martin
PY - 2026
DA - 2026/11/03
JO - PLOS Global Public Health
DO - 10.1371/journal.pgph.0007564
UR - https://journals.plos.org/globalpublichealth/article?id=10.1371/journal.pgph.0007564
AB - The bacterium Chlamydia trachomatis causes three distinct conditions – urogenital chlamydia, the eye disease trachoma, and lymphogranuloma venereum. Use of serological testing for exposure to C. trachomatis for evaluating ocular disease risk factors and population-based surveillance has expanded over the past decade. We developed chimeric monoclonal antibodies to the C. trachomatis antigen Pgp3 as standardized reagents for Pgp3 serological assays. Here we use two anti-Pgp3 monoclonals (1G1 and 1C3) to conduct an interlaboratory comparison of Pgp3 Luminex-based multiplex bead serological assays in 4 different laboratories (each with their own protocol) with 5 operators using 3 different instrument types. Antibodies spotted onto filter paper were stable for two weeks at room temperature. Accuracy and precision were evaluated at 3 concentrations for each monoclonal (high, medium, and low). Accuracy within 20% was achieved for 11/21 conditions for 1G1 and 18/21 conditions for 1C3; the high concentration resulted in the poorest accuracy. Precision was within 10% for all low and medium concentrations and was above 10% for several conditions at the high concentration. The fluorescence values differed between instruments but showed no difference when extrapolated to concentrations using a standard curve. These data suggest that 1C3 may be the more reliable standard reagent and that the Pgp3 multiplex assays are highly robust when comparing different instruments and assay protocols.
SN - 2767-3375
ER -