Title
Assessment of an automated capillary system for Plasmodium vivax microsatellite genotyping
Date Issued
21 August 2015
Access level
open access
Resource Type
journal article
Author(s)
University of California San Diego
Johns Hopkins School of Public Health
Johns Hopkins School of Public Health
Publisher(s)
BioMed Central Ltd.
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<sup>®</sup> 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.
Volume
14
Issue
1
Language
English
OCDE Knowledge area
Epidemiología
Subjects
Scopus EID
2-s2.0-84939643086
PubMed ID
Source
Malaria Journal
ISSN of the container
14752875
Sponsor(s)
We wish to thank Paula Maguina of the University of California for her scientific, logistical and ethics compliance support that were essential for the completion of this work. This work was supported by US Public Health Service Grants U19AI089681, D43TW007120, R01AI067727, and K24AI068903 from the National Institutes of Health (JMV). Medical Research Council - MR/K007467/1
Sources of information:
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