Universidad Peruana Cayetano Heredia

Assessment of an automated capillary system for Plasmodium vivax microsatellite genotyping

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dc.contributor.author Manrique Valverde, Paulo Cesar
dc.contributor.author Hoshi, Mari
dc.contributor.author Fasabi, Manuel
dc.contributor.author Nolasco, Oscar
dc.contributor.author Yori, Pablo
dc.contributor.author Calderón Sánchez, Maritza Mercedes
dc.contributor.author Gilman, Robert Hugh
dc.contributor.author Kosek, Margaret N.
dc.contributor.author Vinetz, Joseph Michael
dc.contributor.author Gamboa Vilela, Dionicia Baziliza
dc.date.accessioned 2019-02-06T14:53:06Z
dc.date.available 2019-02-06T14:53:06Z
dc.date.issued 2015
dc.identifier.uri https://hdl.handle.net/20.500.12866/5364
dc.description.abstract BACKGROUND: Several platforms have been used to generate the primary data for microsatellite analysis of malaria parasite genotypes. Each has relative advantages but share a limitation of being time- and cost-intensive. A commercially available automated capillary gel cartridge system was assessed in the microsatellite analysis of Plasmodium vivax diversity in the Peruvian Amazon. METHODS: The reproducibility and accuracy of a commercially-available automated capillary system, QIAxcel, was assessed using a sequenced PCR product of 227 base pairs. This product was measured 42 times, then 27 P. vivax samples from Peruvian Amazon subjects were analyzed with this instrument using five informative microsatellites. Results from the QIAxcel system were compared with a Sanger-type sequencing machine, the ABI PRISM((R)) 3100 Genetic Analyzer. RESULTS: Significant differences were seen between the sequenced amplicons and the results from the QIAxcel instrument. Different runs, plates and cartridges yielded significantly different results. Additionally, allele size decreased with each run by 0.045, or 1 bp, every three plates. QIAxcel and ABI PRISM systems differed in giving different values than those obtained by ABI PRISM, and too many (i.e. inaccurate) alleles per locus were also seen with the automated instrument. CONCLUSIONS: While P. vivax diversity could generally be estimated using an automated capillary gel cartridge system, the data demonstrate that this system is not sufficiently precise for reliably identifying parasite strains via microsatellite analysis. This conclusion reached after systematic analysis was due both to inadequate precision and poor reproducibility in measuring PCR product size. en_US
dc.language.iso eng
dc.publisher BioMed Central
dc.relation.ispartofseries Malaria Journal
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Humans en_US
dc.subject Peru/epidemiology en_US
dc.subject Reproducibility of Results en_US
dc.subject Microsatellite Repeats/genetics en_US
dc.subject Molecular Epidemiology en_US
dc.subject DNA, Protozoan/genetics en_US
dc.subject Plasmodium vivax/genetics en_US
dc.subject Malaria, Vivax/epidemiology/parasitology en_US
dc.subject Molecular Typing/methods en_US
dc.title Assessment of an automated capillary system for Plasmodium vivax microsatellite genotyping en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1186/s12936-015-0842-9
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.03.07
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.03.08
dc.relation.issn 1475-2875


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