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Pathogens
Volume 10
Issue 2
10.3390/pathogens10020088
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Article

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

by
Navatha Alugubelly
†,
John V. Stokes
†,
Claire E. Cross
,
Anne-Marie L. Ross
,
Anna E. Crawford
,
Gabrielle F. Fiihr
and
Andrea S. Varela-Stokes
*
Department of Comparative Biomedical Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, USA
*
Author to whom correspondence should be addressed.
†
Contributed equally as first authors.
Pathogens 2021, 10(2), 88; https://doi.org/10.3390/pathogens10020088
Submission received: 22 December 2020 / Revised: 18 January 2021 / Accepted: 19 January 2021 / Published: 20 January 2021
(This article belongs to the Collection Updates on Rickettsia and Coxiella)
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Abstract

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.
Keywords: Rickettsia parkeri; Rickettsia amblyommatis; guinea pig; serology; tick-borne diseases; Amblyomma; rickettsial diseases Rickettsia parkeri; Rickettsia amblyommatis; guinea pig; serology; tick-borne diseases; Amblyomma; rickettsial diseases

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MDPI and ACS Style

Alugubelly, N.; Stokes, J.V.; Cross, C.E.; Ross, A.-M.L.; Crawford, A.E.; Fiihr, G.F.; Varela-Stokes, A.S. Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure. Pathogens 2021, 10, 88. https://doi.org/10.3390/pathogens10020088

AMA Style

Alugubelly N, Stokes JV, Cross CE, Ross A-ML, Crawford AE, Fiihr GF, Varela-Stokes AS. Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure. Pathogens. 2021; 10(2):88. https://doi.org/10.3390/pathogens10020088

Chicago/Turabian Style

Alugubelly, Navatha, John V. Stokes, Claire E. Cross, Anne-Marie L. Ross, Anna E. Crawford, Gabrielle F. Fiihr, and Andrea S. Varela-Stokes. 2021. "Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure" Pathogens 10, no. 2: 88. https://doi.org/10.3390/pathogens10020088

APA Style

Alugubelly, N., Stokes, J. V., Cross, C. E., Ross, A.-M. L., Crawford, A. E., Fiihr, G. F., & Varela-Stokes, A. S. (2021). Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure. Pathogens, 10(2), 88. https://doi.org/10.3390/pathogens10020088

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