CRISPR-Cas-based pen-side diagnostic tests for Anaplasma marginale and Babesia bigemina
<i>Anaplasma marginale</i> and <i>Babesia bigemina</i> are tick-borne pathogens, posing significant threats to the health and productivity of cattle in tropical and subtropical regions worldwide. Currently, detection of <i>Babesia bigemina</i> and <i>Anaplasma marginale</i> in infected animals relie...
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| Format: | Journal Article |
| Language: | Inglés |
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MDPI
2024
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| Online Access: | https://hdl.handle.net/10568/168463 |
| _version_ | 1855540675957751808 |
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| author | Muriuki, Robert Ndichu, M. Githigia, S. Svitek, Nicholas |
| author_browse | Githigia, S. Muriuki, Robert Ndichu, M. Svitek, Nicholas |
| author_facet | Muriuki, Robert Ndichu, M. Githigia, S. Svitek, Nicholas |
| author_sort | Muriuki, Robert |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | <i>Anaplasma marginale</i> and <i>Babesia bigemina</i> are tick-borne pathogens, posing significant threats to the health and productivity of cattle in tropical and subtropical regions worldwide. Currently, detection of <i>Babesia bigemina</i> and <i>Anaplasma marginale</i> in infected animals relies primarily on microscopic examination of Giemsa-stained blood or organ smears, which has limited sensitivity. Molecular methods offer higher sensitivity but are costly and impractical in resource-limited settings. Following the development of a pen-side test for detecting <i>Theileria parva</i> infections in cattle, we have created two additional CRISPR-Cas12a assays targeting <i>Anaplasma marginale</i> and <i>Babesia bigemina</i>. The assays target the major surface protein 5 (MSP5) for <i>A. marginale</i> and rhoptry-associated protein 1a (RAP1a) for <i>B. bigemina</i>. These additional tests involve a 20 min recombinase polymerase amplification (RPA) reaction followed by a 60 min CRISPR-Cas12a detection with a lateral strip readout. Results demonstrate high specificity, with no cross-reactivity against other tick-borne parasites, and a limit of detection down to 102 DNA copies/µL of each target marker. The findings pave the way for sensitive and user-friendly pen-side tests to diagnose <i>A. marginale</i> and <i>B. bigemina</i> infections. |
| format | Journal Article |
| id | CGSpace168463 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2024 |
| publishDateRange | 2024 |
| publishDateSort | 2024 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | CGSpace1684632025-12-08T10:29:22Z CRISPR-Cas-based pen-side diagnostic tests for Anaplasma marginale and Babesia bigemina Muriuki, Robert Ndichu, M. Githigia, S. Svitek, Nicholas crispr diagnosis <i>Anaplasma marginale</i> and <i>Babesia bigemina</i> are tick-borne pathogens, posing significant threats to the health and productivity of cattle in tropical and subtropical regions worldwide. Currently, detection of <i>Babesia bigemina</i> and <i>Anaplasma marginale</i> in infected animals relies primarily on microscopic examination of Giemsa-stained blood or organ smears, which has limited sensitivity. Molecular methods offer higher sensitivity but are costly and impractical in resource-limited settings. Following the development of a pen-side test for detecting <i>Theileria parva</i> infections in cattle, we have created two additional CRISPR-Cas12a assays targeting <i>Anaplasma marginale</i> and <i>Babesia bigemina</i>. The assays target the major surface protein 5 (MSP5) for <i>A. marginale</i> and rhoptry-associated protein 1a (RAP1a) for <i>B. bigemina</i>. These additional tests involve a 20 min recombinase polymerase amplification (RPA) reaction followed by a 60 min CRISPR-Cas12a detection with a lateral strip readout. Results demonstrate high specificity, with no cross-reactivity against other tick-borne parasites, and a limit of detection down to 102 DNA copies/µL of each target marker. The findings pave the way for sensitive and user-friendly pen-side tests to diagnose <i>A. marginale</i> and <i>B. bigemina</i> infections. 2024-12-15 2025-01-02T07:32:17Z 2025-01-02T07:32:17Z Journal Article https://hdl.handle.net/10568/168463 en Open Access MDPI Muriuki, R., Ndichu, M., Githigia, S. and Svitek, N. 2024. CRISPR-Cas-based pen-side diagnostic tests for <i>Anaplasma marginale</i> and <i>Babesia bigemina</i>. Microorganisms 12(12): 2595. |
| spellingShingle | crispr diagnosis Muriuki, Robert Ndichu, M. Githigia, S. Svitek, Nicholas CRISPR-Cas-based pen-side diagnostic tests for Anaplasma marginale and Babesia bigemina |
| title | CRISPR-Cas-based pen-side diagnostic tests for Anaplasma marginale and Babesia bigemina |
| title_full | CRISPR-Cas-based pen-side diagnostic tests for Anaplasma marginale and Babesia bigemina |
| title_fullStr | CRISPR-Cas-based pen-side diagnostic tests for Anaplasma marginale and Babesia bigemina |
| title_full_unstemmed | CRISPR-Cas-based pen-side diagnostic tests for Anaplasma marginale and Babesia bigemina |
| title_short | CRISPR-Cas-based pen-side diagnostic tests for Anaplasma marginale and Babesia bigemina |
| title_sort | crispr cas based pen side diagnostic tests for anaplasma marginale and babesia bigemina |
| topic | crispr diagnosis |
| url | https://hdl.handle.net/10568/168463 |
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