Development of a rapid antigen-based lateral flow assay for tick-borne spotted fever rickettsioses.
Tick-borne spotted fever rickettsioses (SFRs) continue to cause severe illness and death in otherwise-healthy individuals due to lack of a timely and reliable diagnostic laboratory test. We recently identified a diagnostic biomarker for SFRs, the putative N-acetylmuramoyl-l-alanine amidase RC0497. H...
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Public Library of Science (PLoS)
2025-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0312819 |
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| author | Richard Willson Yingxin Zhao Kristen Brosamer Yogita Pal Lucas S Blanton Esteban Arroyave Carsen Roach David H Walker Katerina Kourentzi Rong Fang |
| author_facet | Richard Willson Yingxin Zhao Kristen Brosamer Yogita Pal Lucas S Blanton Esteban Arroyave Carsen Roach David H Walker Katerina Kourentzi Rong Fang |
| author_sort | Richard Willson |
| collection | DOAJ |
| description | Tick-borne spotted fever rickettsioses (SFRs) continue to cause severe illness and death in otherwise-healthy individuals due to lack of a timely and reliable diagnostic laboratory test. We recently identified a diagnostic biomarker for SFRs, the putative N-acetylmuramoyl-l-alanine amidase RC0497. Here, we developed a prototype laboratory test that targets RC0497 for diagnosis of SFRs. The concentrations of RC0497 in sera of Rickettsia rickettsii-infected guinea pigs and R. conorii-infected mice were determined by stable isotope dilution-parallel reaction monitoring mass spectrometry (SID-PRM-MS), ranging from 0.1 to 1.1 ng/ml. Using europium chelate nanoparticle reporters, we developed a lateral flow assay (LFA) and evaluated the test with a panel of serum samples of mock and experimentally infected animals. Interestingly, 21 of 22 (95.5%) serum samples from R. rickettsii-infected guinea pigs and R. conorii-infected mice yielded positive results with a ratio of test line / control line greater than the cutoff value determined for non-infected animals. All uninfected samples were in agreement with the intended results, suggesting that the initially assessed specificity of the test is 100%, among these samples. Mice infected with a lethal dose of R. conorii and treated with doxycycline on day 3 post-infection (p.i.), upon RC0497 detection by LFA, displayed significantly decreased rickettsial loads, comparable to the sublethal infection group on day 5 p.i.. A panel of human serum samples spiked with various concentrations of recombinant RC0497 were analyzed by LFA, suggesting that the limit of detection of the LFA was 0.64 ng/mL. These findings suggest that the timely detection of RC0497 by a europium LFA offers guidance for treatment, leading to a significant improvement in infection outcomes. This work, for the first time, shows significant promise for a rapid and easy-to-use platform offering a timely diagnostic assay for severe SFRs. |
| format | Article |
| id | doaj-art-60f7bb85da73497aa4b8e0e9ffd6d3b6 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-60f7bb85da73497aa4b8e0e9ffd6d3b62025-08-20T01:47:28ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01201e031281910.1371/journal.pone.0312819Development of a rapid antigen-based lateral flow assay for tick-borne spotted fever rickettsioses.Richard WillsonYingxin ZhaoKristen BrosamerYogita PalLucas S BlantonEsteban ArroyaveCarsen RoachDavid H WalkerKaterina KourentziRong FangTick-borne spotted fever rickettsioses (SFRs) continue to cause severe illness and death in otherwise-healthy individuals due to lack of a timely and reliable diagnostic laboratory test. We recently identified a diagnostic biomarker for SFRs, the putative N-acetylmuramoyl-l-alanine amidase RC0497. Here, we developed a prototype laboratory test that targets RC0497 for diagnosis of SFRs. The concentrations of RC0497 in sera of Rickettsia rickettsii-infected guinea pigs and R. conorii-infected mice were determined by stable isotope dilution-parallel reaction monitoring mass spectrometry (SID-PRM-MS), ranging from 0.1 to 1.1 ng/ml. Using europium chelate nanoparticle reporters, we developed a lateral flow assay (LFA) and evaluated the test with a panel of serum samples of mock and experimentally infected animals. Interestingly, 21 of 22 (95.5%) serum samples from R. rickettsii-infected guinea pigs and R. conorii-infected mice yielded positive results with a ratio of test line / control line greater than the cutoff value determined for non-infected animals. All uninfected samples were in agreement with the intended results, suggesting that the initially assessed specificity of the test is 100%, among these samples. Mice infected with a lethal dose of R. conorii and treated with doxycycline on day 3 post-infection (p.i.), upon RC0497 detection by LFA, displayed significantly decreased rickettsial loads, comparable to the sublethal infection group on day 5 p.i.. A panel of human serum samples spiked with various concentrations of recombinant RC0497 were analyzed by LFA, suggesting that the limit of detection of the LFA was 0.64 ng/mL. These findings suggest that the timely detection of RC0497 by a europium LFA offers guidance for treatment, leading to a significant improvement in infection outcomes. This work, for the first time, shows significant promise for a rapid and easy-to-use platform offering a timely diagnostic assay for severe SFRs.https://doi.org/10.1371/journal.pone.0312819 |
| spellingShingle | Richard Willson Yingxin Zhao Kristen Brosamer Yogita Pal Lucas S Blanton Esteban Arroyave Carsen Roach David H Walker Katerina Kourentzi Rong Fang Development of a rapid antigen-based lateral flow assay for tick-borne spotted fever rickettsioses. PLoS ONE |
| title | Development of a rapid antigen-based lateral flow assay for tick-borne spotted fever rickettsioses. |
| title_full | Development of a rapid antigen-based lateral flow assay for tick-borne spotted fever rickettsioses. |
| title_fullStr | Development of a rapid antigen-based lateral flow assay for tick-borne spotted fever rickettsioses. |
| title_full_unstemmed | Development of a rapid antigen-based lateral flow assay for tick-borne spotted fever rickettsioses. |
| title_short | Development of a rapid antigen-based lateral flow assay for tick-borne spotted fever rickettsioses. |
| title_sort | development of a rapid antigen based lateral flow assay for tick borne spotted fever rickettsioses |
| url | https://doi.org/10.1371/journal.pone.0312819 |
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