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Journal of Clinical Microbiology
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Immunoassays

Development and Evaluation of a New Triplex Immunoassay That Detects Group A Streptococcus Antibodies for the Diagnosis of Rheumatic Fever

Alana L. Whitcombe, Paulina Hanson-Manful, Susan Jack, Arlo Upton, Polly Atatoa Carr, Deborah A. Williamson, Michael G. Baker, Thomas Proft, Nicole J. Moreland
Yi-Wei Tang, Editor
Alana L. Whitcombe
aSchool of Medical Sciences, The University of Auckland, Auckland, New Zealand
bMaurice Wilkins Centre for Biodiscovery, The University of Auckland, Auckland, New Zealand
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Paulina Hanson-Manful
aSchool of Medical Sciences, The University of Auckland, Auckland, New Zealand
bMaurice Wilkins Centre for Biodiscovery, The University of Auckland, Auckland, New Zealand
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Susan Jack
cSouthern District Health Board, Dunedin, New Zealand
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Arlo Upton
dSouthern Community Laboratories, Dunedin, New Zealand
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Polly Atatoa Carr
eWaikato District Health Board, Hamilton, New Zealand
fWaikato University, Hamilton, New Zealand
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Deborah A. Williamson
gPeter Doherty Institute for Infection and Immunity, Melbourne, VIC, Australia
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  • ORCID record for Deborah A. Williamson
Michael G. Baker
bMaurice Wilkins Centre for Biodiscovery, The University of Auckland, Auckland, New Zealand
hDepartment of Public Health, University of Otago, Wellington, New Zealand
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Thomas Proft
aSchool of Medical Sciences, The University of Auckland, Auckland, New Zealand
bMaurice Wilkins Centre for Biodiscovery, The University of Auckland, Auckland, New Zealand
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Nicole J. Moreland
aSchool of Medical Sciences, The University of Auckland, Auckland, New Zealand
bMaurice Wilkins Centre for Biodiscovery, The University of Auckland, Auckland, New Zealand
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Yi-Wei Tang
Cepheid
Roles: Editor
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DOI: 10.1128/JCM.00300-20
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ABSTRACT

Streptococcal serology is a cornerstone in the diagnosis of acute rheumatic fever (ARF), a postinfectious sequela associated with group A Streptococcus infection. Current tests that measure anti-streptolysin O (ASO) and anti-DNaseB (ADB) titers require parallel processing, with their predictive value limited by the low rate of decay in antibody response. Accordingly, our objective was to develop and assess the diagnostic potential of a triplex bead-based assay, which simultaneously quantifies ASO and ADB together with titers for a third antigen, SpnA. Our previous cytometric bead assay was transferred to the clinically appropriate Luminex platform by coupling streptolysin O, DNaseB, and SpnA to spectrally unique magnetic beads. Sera from more than 350 subjects, including 97 ARF patients, were used to validate the assay and explore immunokinetics. Operating parameters demonstrate that the triplex assay produces accurate and reproducible antibody titers which, for ASO and ADB, are highly correlative with existing assay methodology. When ARF patients were stratified by time (days following hospital admission), there was no difference in ASO and ADB between <28 and 28+ day groups. However, for anti-SpnA, there was a significant decrease (P < 0.05) in the 28+ day group, indicative of faster anti-SpnA antibody decay. Anti-SpnA immunokinetics support very recent group A Streptococcus infection and may assist in diagnostic classification of ARF. Further, bead-based assays enable streptococcal serology to be performed efficiently in a high-throughput manner.

FOOTNOTES

    • Received 18 February 2020.
    • Returned for modification 19 March 2020.
    • Accepted 20 May 2020.
    • Accepted manuscript posted online 27 May 2020.
  • Supplemental material is available online only.

  • Copyright © 2020 American Society for Microbiology.

All Rights Reserved.

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Development and Evaluation of a New Triplex Immunoassay That Detects Group A Streptococcus Antibodies for the Diagnosis of Rheumatic Fever
Alana L. Whitcombe, Paulina Hanson-Manful, Susan Jack, Arlo Upton, Polly Atatoa Carr, Deborah A. Williamson, Michael G. Baker, Thomas Proft, Nicole J. Moreland
Journal of Clinical Microbiology Aug 2020, 58 (9) e00300-20; DOI: 10.1128/JCM.00300-20

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Development and Evaluation of a New Triplex Immunoassay That Detects Group A Streptococcus Antibodies for the Diagnosis of Rheumatic Fever
Alana L. Whitcombe, Paulina Hanson-Manful, Susan Jack, Arlo Upton, Polly Atatoa Carr, Deborah A. Williamson, Michael G. Baker, Thomas Proft, Nicole J. Moreland
Journal of Clinical Microbiology Aug 2020, 58 (9) e00300-20; DOI: 10.1128/JCM.00300-20
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KEYWORDS

immunoassay
DNaseB
SpnA
acute rheumatic fever
group A Streptococcus
immunoassay
serology
streptolysin O

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