Table 2 List of the controls used in each PCR assays for the validation of DNA extraction and/or the real-time PCR runs

PCR HPC

PCR High Positive Control

1

Pf 3D7 DNA extract (QiaAmp DNA mini kit)

0.1 ng/μl

PCR LPC

PCR Low Positive Control

1

Pf 3D7 DNA Extract (QiaAmp DNA mini kit)

0.001 ng/μl

PCR NC

PCR Negative Control

2

H2O

N/A

Ext HPC

Extraction High Positive Control

2

Dried blood spot (4mmØ ) with Pf 3D7 blood

500 parasites/μl

Ext LPC

Extraction Low Positive Control

2

Dried blood spot (4mmØ ) with Pf 3D7 blood

5 parasites/μl

Ext NC

Extraction Negative control

2

Dried blood spot (4mmØ ) with Pf negative blood

No parasite

Pf

P. falciparum Positive Control

1

Plasmid (Pf 400 bp insert)

0.001 ng/μl

Pv

P. vivax Positive Control

1

Plasmid (Pv 400 bp insert)

0.001 ng/μl

Pm

P. malariae Positive Control

1

Plasmid (Pm 400 bp insert)

0.001 ng/μl

Po

P. ovale Positive Control

1

Plasmid (Po 400 bo insert)

0.001 ng/μl