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
cg.contributor.affiliationInternational Center for Tropical Agricultureen
cg.contributor.affiliationUniversidad Nacional de Colombiaen
cg.contributor.donorCGIAR Trust Funden
cg.contributor.initiativeGenebanks
cg.contributor.initiativePlant Health
cg.coverage.countryColombia
cg.coverage.iso3166-alpha2CO
cg.coverage.regionAmericas
cg.coverage.regionSouth America
cg.coverage.regionLatin America and the Caribbean
cg.creator.identifierDiana Patricia Niño Jimenez: 0000-0002-3387-3401en
cg.creator.identifierKarina Lopez-Lopez: 0000-0003-3623-4725en
cg.creator.identifierMaritza Cuervo Ibañez: 0000-0002-4418-699Xen
cg.identifier.doihttps://doi.org/10.1016/j.heliyon.2024.e27604en
cg.isijournalISI Journalen
cg.issn2405-8440en
cg.issue6en
cg.journalHeliyonen
cg.reviewStatusPeer Reviewen
cg.subject.actionAreaGenetic Innovation
cg.subject.alliancebiovciatCASSAVAen
cg.subject.alliancebiovciatHEALTHen
cg.subject.alliancebiovciatPLANT GENETIC RESOURCESen
cg.subject.impactAreaNutrition, health and food security
cg.subject.sdgSDG 2 - Zero hungeren
cg.volume10en
dc.contributor.authorNiño-Jimenez, Diana Patriciaen
dc.contributor.authorLópez-López, Karinaen
dc.contributor.authorCuervo-Ibanez, Maritzaen
dc.date.accessioned2024-04-15T08:12:26Zen
dc.date.available2024-04-15T08:12:26Zen
dc.identifier.urihttps://hdl.handle.net/10568/141463
dc.titleQuantitative detection of cassava common mosaic virus for health certification of cassava (Manihot esculenta Crantz) germplasm using qPCR analysisen
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
dcterms.accessRightsOpen Access
dcterms.available2024-03-04en
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
dcterms.extente27604en
dcterms.issued2024-03-04en
dcterms.languageen
dcterms.licenseCC-BY-NC-4.0
dcterms.publisherElsevieren
dcterms.subjectcscmven
dcterms.subjectqpcren
dcterms.subjectindexingen
dcterms.subjecthealth certificationen
dcterms.subjectdas-elisaen
dcterms.subjectend-point pcren
dcterms.typeJournal Article

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Alliance Bioversity CIAT Journal Articles
Alliance Research Lever 6: Crops for Nutrition and Health
CGIAR Initiative on Genebanks
CGIAR Initiative on Plant Health
CIAT Genetic Resources Articles in International Journals