Absolute Quantification of the Host-To-Parasite DNA Ratio in Theileria parva-Infected Lymphocyte Cell Lines
Theileria parva is a tick-transmitted intracellular apicomplexan pathogen of cattle in sub-Saharan Africa that causes East Coast fever (ECF). ECF is an acute fatal disease that kills over one million cattle annually, imposing a tremendous burden on African small-holder cattle farmers. The pathology...
| Autores principales: | , , , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
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Public Library of Science
2016
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| Acceso en línea: | https://hdl.handle.net/10568/129557 |
| _version_ | 1855524211458572288 |
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| author | Gotia, Hanzel T. Munro, James B. Knowles, Donald P. Daubenberger, Claudia A. Bishop, Richard P. Silva, Joana C. |
| author_browse | Bishop, Richard P. Daubenberger, Claudia A. Gotia, Hanzel T. Knowles, Donald P. Munro, James B. Silva, Joana C. |
| author_facet | Gotia, Hanzel T. Munro, James B. Knowles, Donald P. Daubenberger, Claudia A. Bishop, Richard P. Silva, Joana C. |
| author_sort | Gotia, Hanzel T. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Theileria parva is a tick-transmitted intracellular apicomplexan pathogen of cattle in sub-Saharan Africa that causes East Coast fever (ECF). ECF is an acute fatal disease that kills over one million cattle annually, imposing a tremendous burden on African small-holder cattle farmers. The pathology and level of T. parva infections in its wildlife host, African buffalo (Syncerus caffer), and in cattle are distinct. We have developed an absolute quantification method based on quantitative PCR (qPCR) in which recombinant plasmids containing single copy genes specific to the parasite (apical membrane antigen 1 gene, ama1) or the host (hypoxanthine phosphoribosyltransferase 1, hprt1) are used as the quantification reference standards. Our study shows that T. parva and bovine cells are present in similar numbers in T. parva-infected lymphocyte cell lines and that consequently, due to its much smaller genome size, T. parva DNA comprises between 0.9% and 3% of the total DNA samples extracted from these lines. This absolute quantification assay of parasite and host genome copy number in a sample provides a simple and reliable method of assessing T. parva load in infected bovine lymphocytes, and is accurate over a wide range of host-to-parasite DNA ratios. Knowledge of the proportion of target DNA in a sample, as enabled by this method, is essential for efficient high-throughput genome sequencing applications for a variety of intracellular pathogens. This assay will also be very useful in future studies of interactions of distinct host-T. parva stocks and to fully characterize the dynamics of ECF infection in the field. |
| format | Journal Article |
| id | CGSpace129557 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2016 |
| publishDateRange | 2016 |
| publishDateSort | 2016 |
| publisher | Public Library of Science |
| publisherStr | Public Library of Science |
| record_format | dspace |
| spelling | CGSpace1295572024-08-27T10:35:52Z Absolute Quantification of the Host-To-Parasite DNA Ratio in Theileria parva-Infected Lymphocyte Cell Lines Gotia, Hanzel T. Munro, James B. Knowles, Donald P. Daubenberger, Claudia A. Bishop, Richard P. Silva, Joana C. dna theileria parva theileria parasite lymphocyte Theileria parva is a tick-transmitted intracellular apicomplexan pathogen of cattle in sub-Saharan Africa that causes East Coast fever (ECF). ECF is an acute fatal disease that kills over one million cattle annually, imposing a tremendous burden on African small-holder cattle farmers. The pathology and level of T. parva infections in its wildlife host, African buffalo (Syncerus caffer), and in cattle are distinct. We have developed an absolute quantification method based on quantitative PCR (qPCR) in which recombinant plasmids containing single copy genes specific to the parasite (apical membrane antigen 1 gene, ama1) or the host (hypoxanthine phosphoribosyltransferase 1, hprt1) are used as the quantification reference standards. Our study shows that T. parva and bovine cells are present in similar numbers in T. parva-infected lymphocyte cell lines and that consequently, due to its much smaller genome size, T. parva DNA comprises between 0.9% and 3% of the total DNA samples extracted from these lines. This absolute quantification assay of parasite and host genome copy number in a sample provides a simple and reliable method of assessing T. parva load in infected bovine lymphocytes, and is accurate over a wide range of host-to-parasite DNA ratios. Knowledge of the proportion of target DNA in a sample, as enabled by this method, is essential for efficient high-throughput genome sequencing applications for a variety of intracellular pathogens. This assay will also be very useful in future studies of interactions of distinct host-T. parva stocks and to fully characterize the dynamics of ECF infection in the field. 2016-03-01 2023-03-10T14:39:08Z 2023-03-10T14:39:08Z Journal Article https://hdl.handle.net/10568/129557 en Open Access Public Library of Science Gotia, Hanzel T.; Munro, James B.; Knowles, Donald P.; Daubenberger, Claudia A.; Bishop, Richard P.; Silva, Joana C. 2016. Absolute Quantification of the Host-To-Parasite DNA Ratio in Theileria parva-Infected Lymphocyte Cell Lines. PLOS ONE 11: e0150401 |
| spellingShingle | dna theileria parva theileria parasite lymphocyte Gotia, Hanzel T. Munro, James B. Knowles, Donald P. Daubenberger, Claudia A. Bishop, Richard P. Silva, Joana C. Absolute Quantification of the Host-To-Parasite DNA Ratio in Theileria parva-Infected Lymphocyte Cell Lines |
| title | Absolute Quantification of the Host-To-Parasite DNA Ratio in Theileria parva-Infected Lymphocyte Cell Lines |
| title_full | Absolute Quantification of the Host-To-Parasite DNA Ratio in Theileria parva-Infected Lymphocyte Cell Lines |
| title_fullStr | Absolute Quantification of the Host-To-Parasite DNA Ratio in Theileria parva-Infected Lymphocyte Cell Lines |
| title_full_unstemmed | Absolute Quantification of the Host-To-Parasite DNA Ratio in Theileria parva-Infected Lymphocyte Cell Lines |
| title_short | Absolute Quantification of the Host-To-Parasite DNA Ratio in Theileria parva-Infected Lymphocyte Cell Lines |
| title_sort | absolute quantification of the host to parasite dna ratio in theileria parva infected lymphocyte cell lines |
| topic | dna theileria parva theileria parasite lymphocyte |
| url | https://hdl.handle.net/10568/129557 |
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