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Rapid detection of haemotropic mycoplasma infection of feline erythrocytes using a novel flow cytometric approach
BACKGROUND: The haemotropic mycoplasmas Mycoplasma haemofelis and Candidatus Mycoplasma haemominutum cause feline infectious anaemia with infection rates in feline populations reflecting widespread subclinical infection. Clinically significant infections are much rarer but can be life-threatening. C...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3673821/ https://www.ncbi.nlm.nih.gov/pubmed/23725366 http://dx.doi.org/10.1186/1756-3305-6-158 |
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author | Sánchez-Pérez, Angeles Brown, Graeme Malik, Richard Assinder, Stephen J Cantlon, Katherine Gotsis, Christine Dunbar, Samantha Fraser, Stuart T |
author_facet | Sánchez-Pérez, Angeles Brown, Graeme Malik, Richard Assinder, Stephen J Cantlon, Katherine Gotsis, Christine Dunbar, Samantha Fraser, Stuart T |
author_sort | Sánchez-Pérez, Angeles |
collection | PubMed |
description | BACKGROUND: The haemotropic mycoplasmas Mycoplasma haemofelis and Candidatus Mycoplasma haemominutum cause feline infectious anaemia with infection rates in feline populations reflecting widespread subclinical infection. Clinically significant infections are much rarer but can be life-threatening. Current diagnosis is dependent upon visualising organisms in stained blood smears, PCR or quantitative PCR (qPCR). These procedures are labour-intensive and time-consuming. Furthermore, PCR-based approaches offer limited insight into the disease burden of the infected animal. METHODS: We have developed a novel and rapid flow cytometric system that permits diagnosis of haemotropic mycoplasma infections and quantitation of the percentage of erythrocytes that are parasitized. The method exploits the fact that mature mammalian erythrocytes, the host cell for haemoplasmas, are enucleated and thus lack nucleic acid. DRAQ5 is a synthetic anthrocycline dye which rapidly crosses cell membranes and binds to nucleic acids. The presence of exogenous bacterial DNA in mammalian erythrocytes can, therefore, be detected by DRAQ5 uptake and flow cytometric detection of DRAQ5 fluorescence. RESULTS: Here, we show that this system can detect epi-erythrocytic infection of companion felines by haemotropic mycoplasma. Due to their differences in size, and hence the quantity of DNA, the two major feline hemoplasmas M. haemofelis and Candidatus M. haemominutum can be distinguished according to DRAQ5 fluorescence. We have also shown the usefulness of DRAQ5 uptake in monitoring a cat infected with M. haemofelis sequentially during treatment with doxycycline. CONCLUSIONS: The technique described is the first report of a flow cytometric method for detecting haemotropic mycoplasmas in any species and could be applied to widespread screening of animal populations to assess infection by these epi-erythrocytic parasites. |
format | Online Article Text |
id | pubmed-3673821 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-36738212013-06-06 Rapid detection of haemotropic mycoplasma infection of feline erythrocytes using a novel flow cytometric approach Sánchez-Pérez, Angeles Brown, Graeme Malik, Richard Assinder, Stephen J Cantlon, Katherine Gotsis, Christine Dunbar, Samantha Fraser, Stuart T Parasit Vectors Research BACKGROUND: The haemotropic mycoplasmas Mycoplasma haemofelis and Candidatus Mycoplasma haemominutum cause feline infectious anaemia with infection rates in feline populations reflecting widespread subclinical infection. Clinically significant infections are much rarer but can be life-threatening. Current diagnosis is dependent upon visualising organisms in stained blood smears, PCR or quantitative PCR (qPCR). These procedures are labour-intensive and time-consuming. Furthermore, PCR-based approaches offer limited insight into the disease burden of the infected animal. METHODS: We have developed a novel and rapid flow cytometric system that permits diagnosis of haemotropic mycoplasma infections and quantitation of the percentage of erythrocytes that are parasitized. The method exploits the fact that mature mammalian erythrocytes, the host cell for haemoplasmas, are enucleated and thus lack nucleic acid. DRAQ5 is a synthetic anthrocycline dye which rapidly crosses cell membranes and binds to nucleic acids. The presence of exogenous bacterial DNA in mammalian erythrocytes can, therefore, be detected by DRAQ5 uptake and flow cytometric detection of DRAQ5 fluorescence. RESULTS: Here, we show that this system can detect epi-erythrocytic infection of companion felines by haemotropic mycoplasma. Due to their differences in size, and hence the quantity of DNA, the two major feline hemoplasmas M. haemofelis and Candidatus M. haemominutum can be distinguished according to DRAQ5 fluorescence. We have also shown the usefulness of DRAQ5 uptake in monitoring a cat infected with M. haemofelis sequentially during treatment with doxycycline. CONCLUSIONS: The technique described is the first report of a flow cytometric method for detecting haemotropic mycoplasmas in any species and could be applied to widespread screening of animal populations to assess infection by these epi-erythrocytic parasites. BioMed Central 2013-05-31 /pmc/articles/PMC3673821/ /pubmed/23725366 http://dx.doi.org/10.1186/1756-3305-6-158 Text en Copyright © 2013 Sánchez-Pérez et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Sánchez-Pérez, Angeles Brown, Graeme Malik, Richard Assinder, Stephen J Cantlon, Katherine Gotsis, Christine Dunbar, Samantha Fraser, Stuart T Rapid detection of haemotropic mycoplasma infection of feline erythrocytes using a novel flow cytometric approach |
title | Rapid detection of haemotropic mycoplasma infection of feline erythrocytes using a novel flow cytometric approach |
title_full | Rapid detection of haemotropic mycoplasma infection of feline erythrocytes using a novel flow cytometric approach |
title_fullStr | Rapid detection of haemotropic mycoplasma infection of feline erythrocytes using a novel flow cytometric approach |
title_full_unstemmed | Rapid detection of haemotropic mycoplasma infection of feline erythrocytes using a novel flow cytometric approach |
title_short | Rapid detection of haemotropic mycoplasma infection of feline erythrocytes using a novel flow cytometric approach |
title_sort | rapid detection of haemotropic mycoplasma infection of feline erythrocytes using a novel flow cytometric approach |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3673821/ https://www.ncbi.nlm.nih.gov/pubmed/23725366 http://dx.doi.org/10.1186/1756-3305-6-158 |
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