{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T09:05:25Z","timestamp":1767863125556,"version":"3.49.0"},"reference-count":34,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,11,11]],"date-time":"2022-11-11T00:00:00Z","timestamp":1668124800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003141","name":"Consejo Nacional de Ciencia y Tecnolog\u00eda","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007917","name":"Agricultural Research Service","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007917","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:p>Bovine babesiosis is a tick-borne disease caused by protozoan parasites of the genus <jats:italic>Babesia<\/jats:italic>. <jats:italic>Babesia bigemina<\/jats:italic> is one of the most prevalent and economically important parasite species that infects cattle because of its impact on the meat and milk production industry. Effective disease control strategies should include detection of reservoir animals and early and specific pathogen detection using rapid, economical, sensitive, and specific detection techniques. The loop-mediated isothermal amplification technique (LAMP) is a one-step molecular reaction that amplifies DNA sequences with high sensitivity and specificity under isothermal conditions and requires no special equipment. The results can be observed by the naked eye as color changes. The aim of this work was to develop and standardize the LAMP technique for <jats:italic>B. bigemina<\/jats:italic> detection and its visualization using hydroxynaphtol blue. For this situation, primers were designed from the conserved sequences of the <jats:italic>B. bigemina ama-1<\/jats:italic> gene. The results showed that at 63 \u00b0C in 1 h and under standardized conditions, this technique could amplify <jats:italic>B. bigemina<\/jats:italic> DNA as indicated by the characteristic colorimetric change. Sensitivity evaluation indicated that DNA was amplified at a 0.00000001% parasitemia, and it was demonstrated that this technique specifically amplified the DNA of <jats:italic>B. bigemina<\/jats:italic>. Additionally, this technique could amplify DNA from 10 strains of <jats:italic>B. bigemina<\/jats:italic> from three different countries. It is concluded that the LAMP technique as modified in our case could specifically amplify <jats:italic>B. bigemina<\/jats:italic> DNA and shows high sensitivity, does not cross-react with related organisms, and the product is observed by 60 min of reaction time based on color changes. This report is the first LAMP report that uses sequences that are conserved between strains of the <jats:italic>ama-1<\/jats:italic> gene, demonstrates the results by color changes using hydroxynaphtol blue. We propose LAMP as a rapid and economical alternative method for the molecular detection of <jats:italic>B. bigemina<\/jats:italic>.<\/jats:p>","DOI":"10.3389\/fvets.2022.1056355","type":"journal-article","created":{"date-parts":[[2022,11,11]],"date-time":"2022-11-11T07:06:19Z","timestamp":1668150379000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":7,"title":["Development and standardization of a Loop-mediated isothermal amplification (LAMP) test for the detection of Babesia bigemina"],"prefix":"10.3389","volume":"9","author":[{"given":"Andrea P.","family":"Lizarazo-Zuluaga","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bertha I.","family":"Carvajal-Gamez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Silvina","family":"Wilkowsky","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Silvio","family":"Cravero","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marcos","family":"Trangoni","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juan","family":"Mosqueda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2022,11,11]]},"reference":[{"key":"B1","first-page":"692","article-title":"Sur l'hemoglobinurie bacterienne du boeuf","volume":"107","author":"Babes","year":"1888","journal-title":"CR Acad Sci."},{"key":"B2","doi-asserted-by":"publisher","first-page":"S247","DOI":"10.1017\/S0031182004005190","article-title":"Babesiosis of cattle","volume":"129","author":"Bock","year":"2004","journal-title":"Parasitology"},{"key":"B3","unstructured":"10.5962\/bhl.title.1240681893"},{"key":"B4","doi-asserted-by":"publisher","first-page":"995","DOI":"10.1292\/jvms.12-0425","article-title":"Epidemiological Survey of Babesia bovis and Babesia bigemina Infections of Cattle in Philippines","volume":"75","author":"Yu","year":"2013","journal-title":"J Vet Med Sci."},{"key":"B5","doi-asserted-by":"publisher","first-page":"1788","DOI":"10.1016\/j.meegid.2012.07.004","article-title":"Babesia: a world emerging","author":"Schnittger","year":"2012","journal-title":"Infect Genet."},{"key":"B6","doi-asserted-by":"publisher","first-page":"510","DOI":"10.1111\/j.1749-6632.2000.tb05330.x","article-title":"Babesiosis (Babesia bovis) stability in unstable environments","volume":"916","author":"Smith","year":"2000","journal-title":"Ann N Y Acad Sci."},{"key":"B7","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1016\/j.vetpar.2011.05.032","article-title":"Emerging perspectives in the research of bovine babesiosis and anaplasmosis","volume":"180","author":"Suarez","year":"2011","journal-title":"Vet Parasitol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1111\/j.1365-3024.2005.00748.x","article-title":"Possible mechanisms underlying age-related resistance to bovine babesiosis","volume":"27","author":"Zintl","year":"2005","journal-title":"Parasite Immunol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1016\/S0304-4017(03)00078-5","article-title":"Molecular studies on Babesia, Theileria and Hepatozoon in southern Europe","volume":"113","author":"Criado-Fornelio","year":"2003","journal-title":"Part I Epizootiological aspects Vet Parasitol."},{"key":"B10","doi-asserted-by":"publisher","first-page":"281","DOI":"10.1016\/j.mimet.2007.09.019","article-title":"Development of a multiplex loop-mediated isothermal amplification (mLAMP) method for the simultaneous detection of bovine Babesia parasites","volume":"71","author":"Iseki","year":"2007","journal-title":"J Microbiol Methods."},{"key":"B11","doi-asserted-by":"publisher","first-page":"E63","DOI":"10.1093\/nar\/28.12.e63","article-title":"Loop-mediated isothermal amplification of DNA","volume":"28","author":"Notomi","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"B12","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1006\/mcpr.2002.0415","article-title":"Accelerated reaction by loop-mediated isothermal amplification using loop primers","volume":"16","author":"Nagamine","year":"2002","journal-title":"Mol Cell Probes."},{"key":"B13","doi-asserted-by":"publisher","first-page":"167","DOI":"10.2144\/000113072","article-title":"Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue","volume":"46","author":"Goto","year":"2009","journal-title":"BioTechniques"},{"key":"B14","doi-asserted-by":"publisher","first-page":"407","DOI":"10.1002\/rmv.593","article-title":"Loop mediated isothermal amplification (LAMP): a new generation of innovative gene amplification technique; 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