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A novel loop-mediated isothermal amplification-based test for detecting Neospora caninum DNA

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dc.creatorRamos, Andrea Estefaníaspa
dc.creatorMuñoz, Marinaspa
dc.creatorCortés-Vecino, Jesús Alfredospa
dc.creatorBarato, Paolaspa
dc.creatorPatarroyo, Manuel A.
dc.date.accessioned2020-05-25T23:57:49Z
dc.date.available2020-05-25T23:57:49Z
dc.date.created2017spa
dc.description.abstractBackground: Neospora caninum is a cyst-forming, coccidian parasite which is known to cause neurological disorders in dogs and abortion and neonatal mortality in cows and other livestock. This study reports the development of a loop-mediated isothermal amplification (LAMP) assay based on the Neospora caninum Nc-5 gene and compares its efficacy for detecting DNA to that of a semi-nested PCR test. Results: Six primers were designed based on the Nc-5 repeat region of N. caninum. Specific LAMP primers led to successful amplification of N. caninum DNA at 63 °C in 30 min. The LAMP assay was highly specific (i.e. it did not reveal cross-reactivity with other parasite species) and had a low N. caninum plasmid DNA limit of detection (1 fg), which is ten times higher than that for the semi-nested PCR. LAMP applicability was evaluated using a set of naturally-infected samples (59 from canine faeces and five from bovine abortions). Thirty-nine percent (25/64) of the naturally-infected samples were positive for N. caninum DNA by LAMP and 36% (23/64) by semi-nested PCR. However, the LAMP assay is much faster to perform than semi-nested PCR and provides results in 30 min. Conclusion: The optimized reaction conditions described in this study resulted in a sensitive, specific and rapid technique for detecting N. caninum DNA. Considering the advantages of LAMP for detecting N. caninum DNA, further assays aimed at testing its usefulness on a wider range of field samples are recommended. © 2017 The Author(s).eng
dc.format.mimetypeapplication/pdf
dc.identifier.doihttps://doi.org/10.1186/s13071-017-2549-y
dc.identifier.issn17563305
dc.identifier.urihttps://repository.urosario.edu.co/handle/10336/22749
dc.language.isoengspa
dc.publisherBioMed Central Ltd.spa
dc.relation.citationIssueNo. 1
dc.relation.citationTitleParasites and Vectors
dc.relation.citationVolumeVol. 10
dc.relation.ispartofParasites and Vectors, ISSN:17563305, Vol.10, No.1 (2017)spa
dc.relation.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85036646650&doi=10.1186%2fs13071-017-2549-y&partnerID=40&md5=9c08ad6642204e4759baabc4403b3906spa
dc.rights.accesRightsinfo:eu-repo/semantics/openAccess
dc.rights.accesoAbierto (Texto Completo)spa
dc.source.instnameinstname:Universidad del Rosariospa
dc.source.reponamereponame:Repositorio Institucional EdocURspa
dc.subject.keywordDnaspa
dc.subject.keywordprotozoaneng
dc.subject.keywordPrimer dnaspa
dc.subject.keywordProtozoal dnaspa
dc.subject.keywordArticlespa
dc.subject.keywordControlled studyspa
dc.subject.keywordCryptosporidium parvumspa
dc.subject.keywordDna determinationspa
dc.subject.keywordDna sequencespa
dc.subject.keywordGenespa
dc.subject.keywordGene amplificationspa
dc.subject.keywordHammondia hammondispa
dc.subject.keywordLimit of detectionspa
dc.subject.keywordLoop mediated isothermal amplificationspa
dc.subject.keywordNc 5 genespa
dc.subject.keywordNeospora caninumspa
dc.subject.keywordNeosporosisspa
dc.subject.keywordNonhumanspa
dc.subject.keywordPolymerase chain reactionspa
dc.subject.keywordProtozoonspa
dc.subject.keywordSarcocystisspa
dc.subject.keywordSarcocystis cruzispa
dc.subject.keywordSarcocystis hominisspa
dc.subject.keywordSequence alignmentspa
dc.subject.keywordToxoplasma gondiispa
dc.subject.keywordAnimalspa
dc.subject.keywordBovinespa
dc.subject.keywordCattle diseasespa
dc.subject.keywordCoccidiosisspa
dc.subject.keywordDogspa
dc.subject.keywordDog diseasespa
dc.subject.keywordFecesspa
dc.subject.keywordFemalespa
dc.subject.keywordGeneticsspa
dc.subject.keywordIsolation and purificationspa
dc.subject.keywordNeosporaspa
dc.subject.keywordNucleic acid amplificationspa
dc.subject.keywordParasitologyspa
dc.subject.keywordPregnancyspa
dc.subject.keywordProceduresspa
dc.subject.keywordSensitivity and specificityspa
dc.subject.keywordTemperaturespa
dc.subject.keywordVeterinaryspa
dc.subject.keywordAnimalsspa
dc.subject.keywordAntigenseng
dc.subject.keywordCattlespa
dc.subject.keywordCattle diseasesspa
dc.subject.keywordCoccidiosisspa
dc.subject.keywordDna primersspa
dc.subject.keywordDnaeng
dc.subject.keywordDog diseasesspa
dc.subject.keywordDogsspa
dc.subject.keywordFecesspa
dc.subject.keywordFemalespa
dc.subject.keywordLimit of detectionspa
dc.subject.keywordNeosporaspa
dc.subject.keywordNucleic acid amplification techniquesspa
dc.subject.keywordPolymerase chain reactionspa
dc.subject.keywordPregnancyspa
dc.subject.keywordSensitivity and specificityspa
dc.subject.keywordTemperaturespa
dc.subject.keywordLoop-mediated isothermal amplificationspa
dc.subject.keywordNc-5 genespa
dc.subject.keywordNeospora caninumspa
dc.subject.keywordNeosporosisspa
dc.subject.keywordSemi-nested pcrspa
dc.titleA novel loop-mediated isothermal amplification-based test for detecting Neospora caninum DNAspa
dc.typearticleeng
dc.type.hasVersioninfo:eu-repo/semantics/publishedVersion
dc.type.spaArtículospa
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