New synthetic lipid antigens for rapid serological diagnosis of tuberculosis

Alison Jones; Mark Pitts; Juma’a R. Al Dulayymi; James Gibbons; Andrew Ramsay; Delia Goletti; Christopher D. Gwenin; Mark S. Baird

doi:10.1371/journal.pone.0181414

New synthetic lipid antigens for rapid serological diagnosis of tuberculosis

Alison Jones, Mark Pitts, Juma’a R. Al Dulayymi, James Gibbons, Andrew Ramsay, Delia Goletti, Christopher D. Gwenin, Mark S. Baird^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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Abstract

Background

During pulmonary tuberculosis (PTB) antibodies are generated to trehalose esters of mycolic acids which are cell wall lipids of Mycobacterium tuberculosis (Mtb). Attempts have been made to use these complex natural mixtures in serological tests for PTB diagnosis.

Aim

The aim of this work was to determine whether a serological test based on a panel of defined individual trehalose esters of characteristic synthetic mycolic acids has improved diagnostic accuracy in distinguishing patients with culture positive PTB from individuals who were Mtb culture negative.

Method

One hundred serum samples from well-characterized patients with presumptive tuberculosis, and diagnosed as having pulmonary smear and culture positive TB, or being culture and smear negative were evaluated by ELISA using different combinations of synthetic antigens and secondary antibodies. Using cut-off values determined from these samples, we validated this study blind in samples from a further 249 presumptive TB patients.

Results

With the first 100 samples, detailed responses depended both on the precise structure of the antigen and on the secondary antibody. Using a single antigen, a sensitivity/specificity combination for smear and culture positive PTB detection of 85 and 88% respectively was achieved; this increased to 96% and 95% respectively by a statistical combination of the results with seven antigens. In the blind study a sensitivity/specificity of 87% and 83% was reached with a single antigen. With some synthetic antigens, the responses from all 349 samples were significantly better than those with the natural mixture. Combining the results for seven antigens allowed a distinction between culture positive and negative with a ROC AUC of 0.95.

Conclusion

We have identified promising antigen candidates for serological assays that could be used to diagnose PTB and which could be the basis of a much-needed, simple, rapid diagnostic test that would bring care closer to communities.

Original language	English
Article number	e0181414
Number of pages	28
Journal	PLoS One
Volume	12
Issue number	8
Early online date	14 Aug 2017
DOIs	https://doi.org/10.1371/journal.pone.0181414
Publication status	Published - 14 Aug 2017
Externally published	Yes

Keywords

Enyme-linked immunoassays
Tuberculosis
Mycobacterium tuberculosis
Tuberculosis diagnosis and management
Trehalose
Diagnostic medicine
Antibody response
Esters

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Gwening_NewSynteticLipidAntigens_Published_2017Final published version, 3.85 MBLicence: CC BY

Cite this

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title = "New synthetic lipid antigens for rapid serological diagnosis of tuberculosis",

abstract = "BackgroundDuring pulmonary tuberculosis (PTB) antibodies are generated to trehalose esters of mycolic acids which are cell wall lipids of Mycobacterium tuberculosis (Mtb). Attempts have been made to use these complex natural mixtures in serological tests for PTB diagnosis.AimThe aim of this work was to determine whether a serological test based on a panel of defined individual trehalose esters of characteristic synthetic mycolic acids has improved diagnostic accuracy in distinguishing patients with culture positive PTB from individuals who were Mtb culture negative.MethodOne hundred serum samples from well-characterized patients with presumptive tuberculosis, and diagnosed as having pulmonary smear and culture positive TB, or being culture and smear negative were evaluated by ELISA using different combinations of synthetic antigens and secondary antibodies. Using cut-off values determined from these samples, we validated this study blind in samples from a further 249 presumptive TB patients.ResultsWith the first 100 samples, detailed responses depended both on the precise structure of the antigen and on the secondary antibody. Using a single antigen, a sensitivity/specificity combination for smear and culture positive PTB detection of 85 and 88% respectively was achieved; this increased to 96% and 95% respectively by a statistical combination of the results with seven antigens. In the blind study a sensitivity/specificity of 87% and 83% was reached with a single antigen. With some synthetic antigens, the responses from all 349 samples were significantly better than those with the natural mixture. Combining the results for seven antigens allowed a distinction between culture positive and negative with a ROC AUC of 0.95.ConclusionWe have identified promising antigen candidates for serological assays that could be used to diagnose PTB and which could be the basis of a much-needed, simple, rapid diagnostic test that would bring care closer to communities.",

author = "Alison Jones and Mark Pitts and {Al Dulayymi}, {Juma{\textquoteright}a R.} and James Gibbons and Andrew Ramsay and Delia Goletti and Gwenin, {Christopher D.} and Baird, {Mark S.}",

note = "Funding Information: AJ and MP wish to acknowledge the support of the Welsh Government A4B programme. We wish to thank UNICEF/UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases for providing serum samples from their TB Specimen Bank. Publisher Copyright: {\textcopyright} 2017 Jones et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",

year = "2017",

month = aug,

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doi = "10.1371/journal.pone.0181414",

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T1 - New synthetic lipid antigens for rapid serological diagnosis of tuberculosis

AU - Jones, Alison

AU - Pitts, Mark

AU - Al Dulayymi, Juma’a R.

AU - Gibbons, James

AU - Ramsay, Andrew

AU - Goletti, Delia

AU - Gwenin, Christopher D.

AU - Baird, Mark S.

N1 - Funding Information: AJ and MP wish to acknowledge the support of the Welsh Government A4B programme. We wish to thank UNICEF/UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases for providing serum samples from their TB Specimen Bank. Publisher Copyright: © 2017 Jones et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PY - 2017/8/14

Y1 - 2017/8/14

N2 - BackgroundDuring pulmonary tuberculosis (PTB) antibodies are generated to trehalose esters of mycolic acids which are cell wall lipids of Mycobacterium tuberculosis (Mtb). Attempts have been made to use these complex natural mixtures in serological tests for PTB diagnosis.AimThe aim of this work was to determine whether a serological test based on a panel of defined individual trehalose esters of characteristic synthetic mycolic acids has improved diagnostic accuracy in distinguishing patients with culture positive PTB from individuals who were Mtb culture negative.MethodOne hundred serum samples from well-characterized patients with presumptive tuberculosis, and diagnosed as having pulmonary smear and culture positive TB, or being culture and smear negative were evaluated by ELISA using different combinations of synthetic antigens and secondary antibodies. Using cut-off values determined from these samples, we validated this study blind in samples from a further 249 presumptive TB patients.ResultsWith the first 100 samples, detailed responses depended both on the precise structure of the antigen and on the secondary antibody. Using a single antigen, a sensitivity/specificity combination for smear and culture positive PTB detection of 85 and 88% respectively was achieved; this increased to 96% and 95% respectively by a statistical combination of the results with seven antigens. In the blind study a sensitivity/specificity of 87% and 83% was reached with a single antigen. With some synthetic antigens, the responses from all 349 samples were significantly better than those with the natural mixture. Combining the results for seven antigens allowed a distinction between culture positive and negative with a ROC AUC of 0.95.ConclusionWe have identified promising antigen candidates for serological assays that could be used to diagnose PTB and which could be the basis of a much-needed, simple, rapid diagnostic test that would bring care closer to communities.

AB - BackgroundDuring pulmonary tuberculosis (PTB) antibodies are generated to trehalose esters of mycolic acids which are cell wall lipids of Mycobacterium tuberculosis (Mtb). Attempts have been made to use these complex natural mixtures in serological tests for PTB diagnosis.AimThe aim of this work was to determine whether a serological test based on a panel of defined individual trehalose esters of characteristic synthetic mycolic acids has improved diagnostic accuracy in distinguishing patients with culture positive PTB from individuals who were Mtb culture negative.MethodOne hundred serum samples from well-characterized patients with presumptive tuberculosis, and diagnosed as having pulmonary smear and culture positive TB, or being culture and smear negative were evaluated by ELISA using different combinations of synthetic antigens and secondary antibodies. Using cut-off values determined from these samples, we validated this study blind in samples from a further 249 presumptive TB patients.ResultsWith the first 100 samples, detailed responses depended both on the precise structure of the antigen and on the secondary antibody. Using a single antigen, a sensitivity/specificity combination for smear and culture positive PTB detection of 85 and 88% respectively was achieved; this increased to 96% and 95% respectively by a statistical combination of the results with seven antigens. In the blind study a sensitivity/specificity of 87% and 83% was reached with a single antigen. With some synthetic antigens, the responses from all 349 samples were significantly better than those with the natural mixture. Combining the results for seven antigens allowed a distinction between culture positive and negative with a ROC AUC of 0.95.ConclusionWe have identified promising antigen candidates for serological assays that could be used to diagnose PTB and which could be the basis of a much-needed, simple, rapid diagnostic test that would bring care closer to communities.

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DO - 10.1371/journal.pone.0181414

M3 - Article

C2 - 28806423

AN - SCOPUS:85027325929

SN - 1932-6203

VL - 12

JO - PLoS One

JF - PLoS One

IS - 8

M1 - e0181414

ER -

New synthetic lipid antigens for rapid serological diagnosis of tuberculosis

Abstract

Keywords

UN SDGs

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