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Development and evaluation of an enzyme-linked immunosorbent assay for the detection of antibodies to a common urogenital derivative of Chlamydia trachomatis plasmid-encoded PGP3

Development and evaluation of an enzyme-linked immunosorbent assay for the detection of antibodies to a common urogenital derivative of Chlamydia trachomatis plasmid-encoded PGP3
Development and evaluation of an enzyme-linked immunosorbent assay for the detection of antibodies to a common urogenital derivative of Chlamydia trachomatis plasmid-encoded PGP3

BACKGROUND: Urogenital infection with Chlamydia trachomatis is the most commonly diagnosed sexually transmitted infection in the developed world. Accurate measurement and therefore understanding the seroprevalence of urogenital C. trachomatis infections requires a rigorously optimised and validated ELISA. Previous ELISAs based on the C. trachomatis plasmid-encoded protein, PGP3, have been described but lack standardisation and critical controls or use a less common PGP3 as the capture antigen.

METHODOLOGY/PRINCIPAL FINDINGS: A sensitive and specific indirect ELISA was developed based on recombinant PGP3 derived from a urogenital strain of C. trachomatis, serovar E (pSW2), using a rigorous validation protocol. Serum samples were collected from 166 genitourinary medicine (GUM) clinic patients diagnosed as positive or negative for urogenital C. trachomatis infection by nucleic acid amplification testing (NAATs). Overall sensitivity and specificity compared to NAATs was 68.18% and 98.0%, respectively. Sensitivities for female and male samples were 71.93% and 64.15%, respectively. Comparison of samples from these patients diagnosed positive for C. trachomatis by NAAT and patients diagnosed negative by NAAT revealed statistical significance (p≤0.0001).

CONCLUSIONS: We have developed and validated a sensitive and specific ELISA to detect anti-PGP3 antibodies as an indicator of past and current infection to C. trachomatis using PGP3 from a common urogenital strain. It is anticipated that this assay will be used for seroepidemiological analysis of urogenital C. trachomatis in populations.

Journal Article
0022-1759
Winstanley, Catherine E.
c407b164-5fca-4f53-b13d-e51d2d821489
Ramsey, Kyle H.
b93ffc54-69c6-4bf9-bfe5-cc078ed878fe
Marsh, Peter
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Clarke, Ian N.
ff6c9324-3547-4039-bb2c-10c0b3327a8b
Winstanley, Catherine E.
c407b164-5fca-4f53-b13d-e51d2d821489
Ramsey, Kyle H.
b93ffc54-69c6-4bf9-bfe5-cc078ed878fe
Marsh, Peter
2f77a131-1871-45b8-a5d8-a5ea9beb430a
Clarke, Ian N.
ff6c9324-3547-4039-bb2c-10c0b3327a8b

Winstanley, Catherine E., Ramsey, Kyle H., Marsh, Peter and Clarke, Ian N. (2017) Development and evaluation of an enzyme-linked immunosorbent assay for the detection of antibodies to a common urogenital derivative of Chlamydia trachomatis plasmid-encoded PGP3. Journal of Immunological Methods. (doi:10.1016/j.jim.2017.03.002).

Record type: Article

Abstract

BACKGROUND: Urogenital infection with Chlamydia trachomatis is the most commonly diagnosed sexually transmitted infection in the developed world. Accurate measurement and therefore understanding the seroprevalence of urogenital C. trachomatis infections requires a rigorously optimised and validated ELISA. Previous ELISAs based on the C. trachomatis plasmid-encoded protein, PGP3, have been described but lack standardisation and critical controls or use a less common PGP3 as the capture antigen.

METHODOLOGY/PRINCIPAL FINDINGS: A sensitive and specific indirect ELISA was developed based on recombinant PGP3 derived from a urogenital strain of C. trachomatis, serovar E (pSW2), using a rigorous validation protocol. Serum samples were collected from 166 genitourinary medicine (GUM) clinic patients diagnosed as positive or negative for urogenital C. trachomatis infection by nucleic acid amplification testing (NAATs). Overall sensitivity and specificity compared to NAATs was 68.18% and 98.0%, respectively. Sensitivities for female and male samples were 71.93% and 64.15%, respectively. Comparison of samples from these patients diagnosed positive for C. trachomatis by NAAT and patients diagnosed negative by NAAT revealed statistical significance (p≤0.0001).

CONCLUSIONS: We have developed and validated a sensitive and specific ELISA to detect anti-PGP3 antibodies as an indicator of past and current infection to C. trachomatis using PGP3 from a common urogenital strain. It is anticipated that this assay will be used for seroepidemiological analysis of urogenital C. trachomatis in populations.

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1-s2.0-S0022175916303647-main - Accepted Manuscript
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More information

Accepted/In Press date: 3 March 2017
e-pub ahead of print date: 7 March 2017
Additional Information: Copyright © 2017. Published by Elsevier B.V.
Keywords: Journal Article
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 410809
URI: https://eprints.soton.ac.uk/id/eprint/410809
ISSN: 0022-1759
PURE UUID: 3050423c-c253-4dc6-b825-7b8883777484
ORCID for Ian N. Clarke: ORCID iD orcid.org/0000-0002-4938-1620

Catalogue record

Date deposited: 09 Jun 2017 09:41
Last modified: 10 Dec 2019 06:07

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