Genotyping Malaria Parasites from Blood

Genomic DNA was isolated from whole blood using the QIAamp DNA mini kit (QIAGEN, Valencia, CA). Plasmodium vivax samples were assayed for 25 microsatellites [31 (link),32 (link)] and P. falciparum samples were assayed for 12 microsatellites [33 (link)]. Fluorescently labelled PCR products were separated on an Applied Biosystems 3730 capillary sequencer and scored using GeneMarker v1.95 (SoftGenetics LLC). The finding of one or more additional alleles was interpreted as a co-infection with two or more genetically distinct clones in the same isolate (multiple-clone infection, transmitted by one or several mosquitoes) [33 (link)]. Additional alleles generating peaks of at least one third the height of the predominant allele were also scored. Missing data (no amplifications) are reported by loci but not considered for defining haplotypes.

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Chenet S.M., Schneider K.A., Villegas L, & Escalante A.A. (2012). Local population structure of Plasmodium: impact on malaria control and elimination. Malaria Journal, 11, 412.

Publication 2012

Allele Blood Capillary Clone Co infection Genomic Haplotypes Infection Microsatellites Mosquitoes Plasmodium vivax

Corresponding Organization :

Other organizations : Arizona State University, Hochschule Mittweida

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Protocol cited in 9 other protocols

Variable analysis

independent variables

Plasmodium vivax samples assayed for 25 microsatellites
Plasmodium falciparum samples assayed for 12 microsatellites

dependent variables

Number of alleles found in the microsatellite assay
Presence of co-infections with two or more genetically distinct clones

control variables

Genomic DNA isolation from whole blood using the QIAamp DNA mini kit (QIAGEN, Valencia, CA)
Fluorescently labelled PCR products separated on an Applied Biosystems 3730 capillary sequencer
Scoring of alleles using GeneMarker v1.95 (SoftGenetics LLC)

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