Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia

Table 1 Primers and probe sequences used for QuantiFast™ real-time PCR detection of Plasmodium species [20, 24] and nested PCR detection of P. knowlesi [22] in this study

Species	Primer or probe	Sequence (5′–3′)^a
P. knowlesi
Forward	Plasmo1	GTTAAGGGAGTGAAGACGATCAGA
Reverse	Plasmo2	AACCCAAAGACTTTGATTTCTCATAA
RT-probe	Pkprobe	FAM-CTCTCCGGAGATTAGAACTCTTAGATTGCT-BHQ-1
P. falciparum^b
Forward	Fal-F	CCGACTAGGTGTTGGATGAAAGTGTTAA
RT-probe	Falcprobe	Cy5-AGCAATCTAAAAGTCACCTCGAAAGATGACT-BHQ-1
P. vivax^b
Forward	Viv-F	CCGACTAGGCTTTGGATGAAAGATTTTA
RT-probe	Vivprobe	TR-AGCAATCTAAGAATAAACTCCGAAGAGAAAATTCT-BHQ-2
P. malariae^b
Forward	Mal-F	CCGACTAGGTGTTGGATGATAGAGTAAA
RT-probe	Malaprobe	FAM-CTATCTAAAAGAAACACTCAT-MGBNFQ
Validated reference PCR for P. knowlesi detection
Nest 1; Forward	PkF1160	GATGCCTCCGCGTATCGAC
Nest 1; Reverse	PKR1150	GAGTTCTAATCTCCGGAGAGAAAAGA
Nest 2; Forward	PKF1140	GATTCATCTATTAAAAATTTGCTTC
Nest 2; Reverse	PKR1150	Same as reverse primer used in Nest 1

^aMGBNFQ: minor groove binding non-fluorescent quencher; BHQ: black hole quencher; Cy5: cyanine; FAM: carboxyfluorescein; TR: Texas Red
^bThe Plasmo2 reverse primer was also used for P. falciparum, P. vivax and P. malariae

ISSN: 1475-2875