Quantitative detection of cassava common mosaic virus for health certification of cassava (Manihot esculenta Crantz) germplasm using qPCR analysis

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cg.authorship.typesCGIAR single centreen_US
cg.contributor.affiliationInternational Center for Tropical Agricultureen_US
cg.contributor.affiliationUniversidad Nacional de Colombiaen_US
cg.contributor.donorCGIAR Trust Funden_US
cg.contributor.initiativeGenebanksen_US
cg.contributor.initiativePlant Healthen_US
cg.coverage.countryColombiaen_US
cg.coverage.iso3166-alpha2COen_US
cg.coverage.regionAmericasen_US
cg.coverage.regionSouth Americaen_US
cg.coverage.regionLatin America and the Caribbeanen_US
cg.creator.identifierDiana Patricia Niño Jimenez: 0000-0002-3387-3401en_US
cg.creator.identifierKarina Lopez-Lopez: 0000-0003-3623-4725en_US
cg.creator.identifierMaritza Cuervo Ibañez: 0000-0002-4418-699Xen_US
cg.identifier.doihttps://doi.org/10.1016/j.heliyon.2024.e27604en_US
cg.isijournalISI Journalen_US
cg.issn2405-8440en_US
cg.issue6en_US
cg.journalHeliyonen_US
cg.reviewStatusPeer Reviewen_US
cg.subject.actionAreaGenetic Innovationen_US
cg.subject.alliancebiovciatCASSAVAen_US
cg.subject.alliancebiovciatHEALTHen_US
cg.subject.alliancebiovciatPLANT GENETIC RESOURCESen_US
cg.subject.impactAreaNutrition, health and food securityen_US
cg.subject.sdgSDG 2 - Zero hungeren_US
cg.volume10en_US
dc.contributor.authorNiño-Jimenez, Diana Patriciaen_US
dc.contributor.authorLópez-López, Karinaen_US
dc.contributor.authorCuervo-Ibanez, Maritzaen_US
dc.date.accessioned2024-04-15T08:12:26Zen_US
dc.date.available2024-04-15T08:12:26Zen_US
dc.identifier.urihttps://hdl.handle.net/10568/141463en_US
dc.titleQuantitative detection of cassava common mosaic virus for health certification of cassava (Manihot esculenta Crantz) germplasm using qPCR analysisen_US
dcterms.abstractCassava (Manihot esculenta Crantz) is a crop of global economic and food safety importance, used for human consumption and in various industrial applications. The genebank of the Genetic Resources Program of the Alliance of Bioversity International and CIAT currently holds the world's largest cassava collection, with 5965 <i>in vitro</i> accessions from 28 countries. Managing this extensive collection involves indexing quarantine pathogens as a phytosanitary certification requirement for safely distributing cassava germplasm. The study therefore aimed to optimize a quantitative diagnostic protocol to detect cassava common mosaic virus (CsCMV) using quantitative PCR (qPCR) as a better alternative to other molecular techniques. This was done through designing primers and a probe in the RdRP region of CsCMV, and optimizing the qPCR conditions of the diagnostic protocol using primer concentration assays, and reaction amplification conditions such as volume and reaction time. We also evaluated the qPCR protocol by comparing the results of 140 cassava accession evaluations using three diagnostic methodologies (DAS-ELISA, end-point PCR, and qPCR) for CsCMV. Our protocol established that qPCR technique analysis is ten-times more sensitive in detecting CsCMV compared to end-point PCR, showing a maximum detection level of 77.97 copies/μL of plasmid, with 76 min of reaction time. The comparison allowed us to verify the level of CsCMV detection through the techniques evaluated, concluding that qPCR was more sensitive and allowed the quantification of viral concentration. The optimized qPCR protocol will be used to accelerate diagnostic screening of cassava germplasm for the presence or absence of CsCMV to ensure safe movement and distribution of disease-free germplasm.en_US
dcterms.accessRightsOpen Accessen_US
dcterms.available2024-03-04en_US
dcterms.bibliographicCitationNiño-Jimenez, D.P.; López-López, K.; Cuervo-Ibanez, M. (2024) Quantitative detection of cassava common mosaic virus for health certification of cassava (Manihot esculenta Crantz) germplasm using qPCR analysis. Heliyon 10(6): e27604. ISSN: 2405-8440en_US
dcterms.extente27604en_US
dcterms.issued2024-03-04en_US
dcterms.languageenen_US
dcterms.licenseCC-BY-NC-4.0en_US
dcterms.publisherElsevieren_US
dcterms.subjectcscmven_US
dcterms.subjectqpcren_US
dcterms.subjectindexingen_US
dcterms.subjecthealth certificationen_US
dcterms.subjectdas-elisaen_US
dcterms.subjectend-point pcren_US
dcterms.typeJournal Articleen_US

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CGIAR Initiative on Genebanks
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