Diagnosis of an imported Plasmodium ovale wallikeri infection in Malaysia

Liew, Jonathan Wee Kent; Mahmud, Rohela; Tan, Lian Huat; Lau, Yee Ling

doi:10.1186/s12936-015-1070-z

Malaria Journal

Table 1 Description of PCR protocols used in this report

From: Diagnosis of an imported Plasmodium ovale wallikeri infection in Malaysia

Reaction	Primer	Annealing temperature (°C)	Product size (bp)	Target species	Reference
Nested PCR
Nest 1^a	rPLU1/rPLU5	55	≈1670	Plasmodium spp.	[19]
Nest 2^b	rPLU3/rPLU4		240	Plasmodium spp.
	rFal1/rFal2		205	P. falciparum
	rVIV1/rVIV2		120	P. vivax
	rMAL1/rMAL2		145	P. malariae
	rOVA1/rOVA2	58	787–789	P. ovale curtisi
	Pmk8/Pmkr9		153	P. knowlesi	[16]
	rOVA1v/rOVA2v		782	P. ovale wallikeri	[3]
	rOVA1WC/rOVA2WC		659–662	P. ovale curtisi and P. ovale wallikeri	[26]
Mixed P. ovale ^c	rOVA1/rOVA2 and rOVA1v/rOVA2v	58	782–789	P. ovale curtisi and P. ovale wallikeri	[3]
NM- and NG-PCR
Primary PCR^d	UNR/PLF	58	783–821	Universal	[20]
NM PCR^e	New PLFshort with	53		Plasmodium spp.
	MARshort		241	P. malariae
	FARshort		370	P. falciparum
	OVRshort		407	P. ovale
	VIRshort		476	P. vivax
NG-PCR^f	NewPLFshort/NewRevshort	53	735–773	Plasmodium spp.

^aCycling condition: Initial denaturation, 94 °C for 4 min; 35 cycles of 94 °C for 1 min, 55 °C for 1 min and 72 °C for 1 min; final extension 72 °C for 10 min. Four µL of DNA is used in a 25 µL reaction volume containing 4 mM magnesium chloride, 0.2 mM of each dNTPs and 1 Unit of Taq polymerase
^bCycling condition: Initial denaturation, 94 °C for 4 min; 35 cycles of 94 °C for 1 min, 58 °C for 1 min and 72 °C for 1 min; final extension 72 °C for 10 min. Four µL of nest one product is used in a 25 µL reaction volume containing 4 mM magnesium chloride, 0.2 mM of each dNTPs and 1 Unit of Taq polymerase
^cCycling condition is the same as Nest 2 PCR^b. Primer sets rOVA1/rOVA2 and rOVA1v/rOVA2v are mixed in equimolar in a single PCR reaction tube. Four µL of nest one product is used in a 25 µL reaction volume containing 4 mM magnesium chloride, 0.2 mM of each dNTPs and 1 Unit of Taq polymerase
^dCycling condition: Initial denaturation, 94 °C for 5 min; 35 cycles of 94 °C for 1 min, 58 °C for 1 min and 72 °C for 1 min; final extension 72 °C for 10 min. Five µL of DNA is used in a 50 µL reaction volume containing 4 mM magnesium chloride, 0.2 mM of each dNTPs and 2 Units of Taq polymerase
^eCycling condition: Initial denaturation, 94 °C for 5 min; 35 cycles of 94 °C for 1 min, 53 °C for 1 min and 72 °C for 1 min; final extension 72 °C for 10 min. Two µL of Primary PCR product is used in a 25 µL reaction volume containing 4 mM magnesium chloride, 0.2 mM of each dNTPs and 1 Unit of Taq polymerase
^fCycling condition is the same as NM-PCR^e. Two µL of Primary PCR product is used in a 50 µL reaction volume containing 4 mM magnesium chloride, 0.2 mM of each dNTPs and 2 Units of Taq polymerase

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ISSN: 1475-2875

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