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Volume 12, Number 3—March 2006

Research

Identifying and Quantifying Genotypes in Polyclonal Infections due to Single Species

James M. Colborn*, Ousmane Koita†, Ousmane Cissé†, Mamadou W. Bagayoko†, Edward J. Guthrie‡, and Donald J. Krogstad*Comments to Author 
Author affiliations: *Tulane University Health Sciences Center, New Orleans, Louisiana, USA; †University of Bamako, Bamako, Mali; ‡Agilent Technologies, Wilmington, Delaware, USA

Main Article

Table 1

Primers and probes used for real-time polymerase chain reaction*

Primer/probe sequence Sizes (bp) Tm (°C) Fluorophore, quencher Final reactant concentration (nmol/L)
Parasites with K1 and hybrid sequences in block 2 of msp1
K1F 5´-AGGTGCAAGTGCTCAAAGTG-3´ 108–171 50.3 Texas Red, BHQ-2 100
K1R 5´-CACCAGATGAAGTATTTGAACG-3´ 49.2 100
PROBE: 5´-AAGTGGTACAAGTCCATCATCTCGT-3´ 54.9 300
Hybrid F 5´-GAAGGAACAAGTGGAACAGC-3´ 96–168 48.7 Cy5, BHQ-2 200
Hybrid R 5´-GCAGCACCTGGAGATCTTATA-3´ 48.8 200
PROBE: 5´-TTCACTTATTTCCCCATGAGCCCC-3´ 54.2 400
Primers for positive control (EBA175)
EBA175 F 5´-GGTTATTCAACTAAGGCAGAA-3´ 95 46.0 Cy 5, BHQ-3 100
EBA175 R 5´-TCCACCATTCTTTTCTAAAATTTT-3´ 50.6 100
PROBE: 5´-TCATTTCCCATAGCAAGATGTCC 60.0 100
LUX primers (MAD 20, RO33)
MAD20 F 5´-AATGAAGGAACAAGTGGAAC-3´ 52–205 48.7 FAM 200
MAD20 R 5´-GAATTATCTGAAGGATTTGTACG-3´ 47.4 (reverse) 200
RO33 F 5´-GCAGATGCTGTAAGTACTCAA-3´ 148 44.8 JOE 200
RO33 R 5´-GCAGCACCTGGAGATCTTATA-3´ 44.8 (forward) 200

*msp1, merozoite surface protein 1, EBA175, erythrocyte-binding antigen 175; Tm, melting temperature.

Main Article

Page created: January 27, 2012
Page updated: January 27, 2012
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