RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infection
Abstract Anaplasma marginale infection is one of the most common tick-borne diseases, causing a substantial loss in the beef and dairy production industries. Once infected, the pathogen remains in the cattle for life, allowing the parasites to spread to healthy animals. Since clinical manifestations...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-04-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-58169-6 |
| _version_ | 1827291597505560576 |
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| author | Arpaporn Sutipatanasomboon Jantana Wongsantichon Somsri Sakdee Piyaporn Naksith Amaya Watthanadirek Panat Anuracpreeda Stuart D. Blacksell Chonticha Saisawang |
| author_facet | Arpaporn Sutipatanasomboon Jantana Wongsantichon Somsri Sakdee Piyaporn Naksith Amaya Watthanadirek Panat Anuracpreeda Stuart D. Blacksell Chonticha Saisawang |
| author_sort | Arpaporn Sutipatanasomboon |
| collection | DOAJ |
| description | Abstract Anaplasma marginale infection is one of the most common tick-borne diseases, causing a substantial loss in the beef and dairy production industries. Once infected, the pathogen remains in the cattle for life, allowing the parasites to spread to healthy animals. Since clinical manifestations of anaplasmosis occur late in the disease, a sensitive, accurate, and affordable pathogen identification is crucial in preventing and controlling the infection. To this end, we developed an RPA-CRISPR/Cas12a assay specific to A. marginale infection in bovines targeting the msp4 gene. Our assay is performed at one moderately high temperature, producing fluorescent signals or positive readout of a lateral flow dipstick, which is as sensitive as conventional PCR-based DNA amplification. This RPA-CRISPR/Cas12a assay can detect as few as 4 copies/μl of Anaplasma using msp4 marker without cross-reactivity to other common bovine pathogens. Lyophilized components of the assay can be stored at room temperature for an extended period, indicating its potential for field diagnosis and low-resource settings of anaplasmosis in bovines. |
| first_indexed | 2024-04-24T12:39:37Z |
| format | Article |
| id | doaj.art-9bdb071be2414d03b1da6db96e6b24ae |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-24T12:39:37Z |
| publishDate | 2024-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-9bdb071be2414d03b1da6db96e6b24ae2024-04-07T11:17:55ZengNature PortfolioScientific Reports2045-23222024-04-0114111110.1038/s41598-024-58169-6RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infectionArpaporn Sutipatanasomboon0Jantana Wongsantichon1Somsri Sakdee2Piyaporn Naksith3Amaya Watthanadirek4Panat Anuracpreeda5Stuart D. Blacksell6Chonticha Saisawang7Molecular Biosciences Cluster, Institute of Molecular Biosciences, Mahidol University, Salaya CampusMahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol UniversityCenter for Advanced Therapeutics, Institute of Molecular Biosciences, Mahidol University, Salaya CampusCenter for Advanced Therapeutics, Institute of Molecular Biosciences, Mahidol University, Salaya CampusMolecular Biosciences Cluster, Institute of Molecular Biosciences, Mahidol University, Salaya CampusMolecular Biosciences Cluster, Institute of Molecular Biosciences, Mahidol University, Salaya CampusMahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol UniversityCenter for Advanced Therapeutics, Institute of Molecular Biosciences, Mahidol University, Salaya CampusAbstract Anaplasma marginale infection is one of the most common tick-borne diseases, causing a substantial loss in the beef and dairy production industries. Once infected, the pathogen remains in the cattle for life, allowing the parasites to spread to healthy animals. Since clinical manifestations of anaplasmosis occur late in the disease, a sensitive, accurate, and affordable pathogen identification is crucial in preventing and controlling the infection. To this end, we developed an RPA-CRISPR/Cas12a assay specific to A. marginale infection in bovines targeting the msp4 gene. Our assay is performed at one moderately high temperature, producing fluorescent signals or positive readout of a lateral flow dipstick, which is as sensitive as conventional PCR-based DNA amplification. This RPA-CRISPR/Cas12a assay can detect as few as 4 copies/μl of Anaplasma using msp4 marker without cross-reactivity to other common bovine pathogens. Lyophilized components of the assay can be stored at room temperature for an extended period, indicating its potential for field diagnosis and low-resource settings of anaplasmosis in bovines.https://doi.org/10.1038/s41598-024-58169-6 |
| spellingShingle | Arpaporn Sutipatanasomboon Jantana Wongsantichon Somsri Sakdee Piyaporn Naksith Amaya Watthanadirek Panat Anuracpreeda Stuart D. Blacksell Chonticha Saisawang RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infection Scientific Reports |
| title | RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infection |
| title_full | RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infection |
| title_fullStr | RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infection |
| title_full_unstemmed | RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infection |
| title_short | RPA-CRISPR/Cas12a assay for the diagnosis of bovine Anaplasma marginale infection |
| title_sort | rpa crispr cas12a assay for the diagnosis of bovine anaplasma marginale infection |
| url | https://doi.org/10.1038/s41598-024-58169-6 |
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