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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 1a rPLU1/rPLU5 55 ≈1670 Plasmodium spp. [19]
Nest 2b 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 PCRd UNR/PLF 58 783–821 Universal [20]
NM
PCRe
New PLFshort with 53   Plasmodium spp.
MARshort   241 P. malariae
FARshort   370 P. falciparum
OVRshort   407 P. ovale
VIRshort   476 P. vivax
NG-PCRf NewPLFshort/NewRevshort 53 735–773 Plasmodium spp.
  1. 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
  2. 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
  3. cCycling condition is the same as Nest 2 PCRb. 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
  4. 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
  5. 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
  6. fCycling condition is the same as NM-PCRe. 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