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Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure

Based on limited serological studies, at least 10% of the US population has been exposed to spotted fever group Rickettsia (SFGR) species. The immunofluorescence antibody assay (IFA) has been the gold standard for the serodiagnosis of rickettsial infections such as spotted fever rickettsiosis (SFR)....

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Autores principales: Alugubelly, Navatha, Stokes, John V., Cross, Claire E., Ross, Anne-Marie L., Crawford, Anna E., Fiihr, Gabrielle F., Varela-Stokes, Andrea S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7909427/
https://www.ncbi.nlm.nih.gov/pubmed/33498380
http://dx.doi.org/10.3390/pathogens10020088
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author Alugubelly, Navatha
Stokes, John V.
Cross, Claire E.
Ross, Anne-Marie L.
Crawford, Anna E.
Fiihr, Gabrielle F.
Varela-Stokes, Andrea S.
author_facet Alugubelly, Navatha
Stokes, John V.
Cross, Claire E.
Ross, Anne-Marie L.
Crawford, Anna E.
Fiihr, Gabrielle F.
Varela-Stokes, Andrea S.
author_sort Alugubelly, Navatha
collection PubMed
description Based on limited serological studies, at least 10% of the US population has been exposed to spotted fever group Rickettsia (SFGR) species. The immunofluorescence antibody assay (IFA) has been the gold standard for the serodiagnosis of rickettsial infections such as spotted fever rickettsiosis (SFR). However, the IFA is semi-quantitative and subjective, requiring a high level of expertise to interpret it correctly. Here, we developed an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of Rickettsia parkeri infection in the guinea pig. Our ELISA is an objective, quantitative, and high-throughput assay that shows greater sensitivity and resolution in observed titers than the IFA. We methodically optimized relevant parameters in sequence for optimal signal-to-noise ratio and low coefficient of variation% values. We used a guinea pig model as it is a part of our overall research efforts to understand the immunological and clinical response to SFGR species after tick transmission. Guinea pigs are a useful model to study SFR and show clinical signs of SFR, such as fever and eschars. We anticipate that this assay will be easily adapted to other hosts, including humans and other SFGR species.
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spelling pubmed-79094272021-02-27 Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure Alugubelly, Navatha Stokes, John V. Cross, Claire E. Ross, Anne-Marie L. Crawford, Anna E. Fiihr, Gabrielle F. Varela-Stokes, Andrea S. Pathogens Article Based on limited serological studies, at least 10% of the US population has been exposed to spotted fever group Rickettsia (SFGR) species. The immunofluorescence antibody assay (IFA) has been the gold standard for the serodiagnosis of rickettsial infections such as spotted fever rickettsiosis (SFR). However, the IFA is semi-quantitative and subjective, requiring a high level of expertise to interpret it correctly. Here, we developed an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of Rickettsia parkeri infection in the guinea pig. Our ELISA is an objective, quantitative, and high-throughput assay that shows greater sensitivity and resolution in observed titers than the IFA. We methodically optimized relevant parameters in sequence for optimal signal-to-noise ratio and low coefficient of variation% values. We used a guinea pig model as it is a part of our overall research efforts to understand the immunological and clinical response to SFGR species after tick transmission. Guinea pigs are a useful model to study SFR and show clinical signs of SFR, such as fever and eschars. We anticipate that this assay will be easily adapted to other hosts, including humans and other SFGR species. MDPI 2021-01-20 /pmc/articles/PMC7909427/ /pubmed/33498380 http://dx.doi.org/10.3390/pathogens10020088 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Alugubelly, Navatha
Stokes, John V.
Cross, Claire E.
Ross, Anne-Marie L.
Crawford, Anna E.
Fiihr, Gabrielle F.
Varela-Stokes, Andrea S.
Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure
title Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure
title_full Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure
title_fullStr Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure
title_full_unstemmed Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure
title_short Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure
title_sort beyond the ifa: revisiting the elisa as a more sensitive, objective, and quantitative evaluation of spotted fever group rickettsia exposure
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7909427/
https://www.ncbi.nlm.nih.gov/pubmed/33498380
http://dx.doi.org/10.3390/pathogens10020088
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