Cargando…

A Serodiagnostic IgM ELISA to Detect Acute Cytauxzoonosis

Cytauxzoonosis is a tick-borne infectious disease affecting domestic cats with high mortality and limited treatment modalities. Because early diagnosis and therapeutic intervention are crucial to survival of infected cats, the objective of this study was to develop an ELISA capable of detecting cyta...

Descripción completa

Detalles Bibliográficos
Autores principales: Kao, Yun-Fan, Spainhour, Rebecca, Cowan, Shannon R., Nafe, Laura, Birkenheuer, Adam, Reichard, Mason V., Miller, Craig A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9611129/
https://www.ncbi.nlm.nih.gov/pubmed/36297239
http://dx.doi.org/10.3390/pathogens11101183
_version_ 1784819449792036864
author Kao, Yun-Fan
Spainhour, Rebecca
Cowan, Shannon R.
Nafe, Laura
Birkenheuer, Adam
Reichard, Mason V.
Miller, Craig A.
author_facet Kao, Yun-Fan
Spainhour, Rebecca
Cowan, Shannon R.
Nafe, Laura
Birkenheuer, Adam
Reichard, Mason V.
Miller, Craig A.
author_sort Kao, Yun-Fan
collection PubMed
description Cytauxzoonosis is a tick-borne infectious disease affecting domestic cats with high mortality and limited treatment modalities. Because early diagnosis and therapeutic intervention are crucial to survival of infected cats, the objective of this study was to develop an ELISA capable of detecting cytauxzoonosis and differentiating acute vs. chronic infection in clinical feline blood samples. A microsphere immunoassay (MIA) was developed to evaluate the production of Cytauxzoon felis-specific IgM and IgG antibodies in serial plasma samples from cats with experimental C. felis infection by targeting a C. felis-specific transmembrane protein (c88). Recombinant c88 protein was utilized to develop indirect ELISAs to detect IgM and IgG antibodies in clinical plasma samples from: PCR-positive cats with acute C. felis infection (n = 36), C. felis-negative cats with pyrexia (n = 10), healthy C. felis-negative cats (n = 22), and chronic C. felis carriers (n = 4). Anti-c88 IgM antibodies were detectable at day 12 post-tick infestation in cats with experimental C. felis infection (within 24 hours of developing clinical signs), while anti-c88 IgG was detectable at day 15 post-tick infestation – indicating IgM could be used to detect early infection. Using a cut-off value of 19.85 percent positive, the C. felis IgM ELISA detected acute cytauxzoonosis in 94.44% (34/36) of cats presented with clinical signs of acute cytauxzoonosis with 100% specificity (indicating a “Strong Positive” result). When a lower cutoff of 8.60 percent positive was used, cytauxzoonosis was detected in the 2 remaining PCR-positive cats with 87.88% specificity (indicating of a “Weak Positive” result). One C. felis-negative, febrile cat had high IgG, and chronic carriers had variable IgM and IgG results. Combined interpretation of IgM and IgG ELISAs did not reliably differentiate acute vs. chronic infection. While further validation on assay performance is needed, the C. felis IgM ELISA is a promising test to detect acute cytauxzoonosis and can be utilized to develop a point-of-care test for clinical use.
format Online
Article
Text
id pubmed-9611129
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-96111292022-10-28 A Serodiagnostic IgM ELISA to Detect Acute Cytauxzoonosis Kao, Yun-Fan Spainhour, Rebecca Cowan, Shannon R. Nafe, Laura Birkenheuer, Adam Reichard, Mason V. Miller, Craig A. Pathogens Article Cytauxzoonosis is a tick-borne infectious disease affecting domestic cats with high mortality and limited treatment modalities. Because early diagnosis and therapeutic intervention are crucial to survival of infected cats, the objective of this study was to develop an ELISA capable of detecting cytauxzoonosis and differentiating acute vs. chronic infection in clinical feline blood samples. A microsphere immunoassay (MIA) was developed to evaluate the production of Cytauxzoon felis-specific IgM and IgG antibodies in serial plasma samples from cats with experimental C. felis infection by targeting a C. felis-specific transmembrane protein (c88). Recombinant c88 protein was utilized to develop indirect ELISAs to detect IgM and IgG antibodies in clinical plasma samples from: PCR-positive cats with acute C. felis infection (n = 36), C. felis-negative cats with pyrexia (n = 10), healthy C. felis-negative cats (n = 22), and chronic C. felis carriers (n = 4). Anti-c88 IgM antibodies were detectable at day 12 post-tick infestation in cats with experimental C. felis infection (within 24 hours of developing clinical signs), while anti-c88 IgG was detectable at day 15 post-tick infestation – indicating IgM could be used to detect early infection. Using a cut-off value of 19.85 percent positive, the C. felis IgM ELISA detected acute cytauxzoonosis in 94.44% (34/36) of cats presented with clinical signs of acute cytauxzoonosis with 100% specificity (indicating a “Strong Positive” result). When a lower cutoff of 8.60 percent positive was used, cytauxzoonosis was detected in the 2 remaining PCR-positive cats with 87.88% specificity (indicating of a “Weak Positive” result). One C. felis-negative, febrile cat had high IgG, and chronic carriers had variable IgM and IgG results. Combined interpretation of IgM and IgG ELISAs did not reliably differentiate acute vs. chronic infection. While further validation on assay performance is needed, the C. felis IgM ELISA is a promising test to detect acute cytauxzoonosis and can be utilized to develop a point-of-care test for clinical use. MDPI 2022-10-14 /pmc/articles/PMC9611129/ /pubmed/36297239 http://dx.doi.org/10.3390/pathogens11101183 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kao, Yun-Fan
Spainhour, Rebecca
Cowan, Shannon R.
Nafe, Laura
Birkenheuer, Adam
Reichard, Mason V.
Miller, Craig A.
A Serodiagnostic IgM ELISA to Detect Acute Cytauxzoonosis
title A Serodiagnostic IgM ELISA to Detect Acute Cytauxzoonosis
title_full A Serodiagnostic IgM ELISA to Detect Acute Cytauxzoonosis
title_fullStr A Serodiagnostic IgM ELISA to Detect Acute Cytauxzoonosis
title_full_unstemmed A Serodiagnostic IgM ELISA to Detect Acute Cytauxzoonosis
title_short A Serodiagnostic IgM ELISA to Detect Acute Cytauxzoonosis
title_sort serodiagnostic igm elisa to detect acute cytauxzoonosis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9611129/
https://www.ncbi.nlm.nih.gov/pubmed/36297239
http://dx.doi.org/10.3390/pathogens11101183
work_keys_str_mv AT kaoyunfan aserodiagnosticigmelisatodetectacutecytauxzoonosis
AT spainhourrebecca aserodiagnosticigmelisatodetectacutecytauxzoonosis
AT cowanshannonr aserodiagnosticigmelisatodetectacutecytauxzoonosis
AT nafelaura aserodiagnosticigmelisatodetectacutecytauxzoonosis
AT birkenheueradam aserodiagnosticigmelisatodetectacutecytauxzoonosis
AT reichardmasonv aserodiagnosticigmelisatodetectacutecytauxzoonosis
AT millercraiga aserodiagnosticigmelisatodetectacutecytauxzoonosis
AT kaoyunfan serodiagnosticigmelisatodetectacutecytauxzoonosis
AT spainhourrebecca serodiagnosticigmelisatodetectacutecytauxzoonosis
AT cowanshannonr serodiagnosticigmelisatodetectacutecytauxzoonosis
AT nafelaura serodiagnosticigmelisatodetectacutecytauxzoonosis
AT birkenheueradam serodiagnosticigmelisatodetectacutecytauxzoonosis
AT reichardmasonv serodiagnosticigmelisatodetectacutecytauxzoonosis
AT millercraiga serodiagnosticigmelisatodetectacutecytauxzoonosis