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Table 1 Description of NM-PCR and NG-PCR with the corresponding primers sequences, concentration and annealing temperature

From: First case of a naturally acquired human infection with Plasmodium cynomolgi

Reaction

Primer name

Primer (μmol/L)

Sequence (5’-3’)

Annealing temp. (°C)

Product size (bp)

Specificity

First PCR*

UNR

0.10

GACGGTATCTGATCGTCTTC

58°C

 

Universal

PLF

0.10

AGTGTGTATCCAATCGAGTTTC

783–821§

Plasmodium

HUF

0.01

GAGCCGCCTGGATACCGC

231

Human

NM-PCR†

NewPLFshort

0.15

CTATCAGCTTTTGATGTTAG

53°C

 

Plasmodium

MARshort

0.10

TCCAATTGCCTTCTG

241

P. malariae

FARshort

0.15

GTTCCCCTAGAATAGTTACA

370

P. falciparum

OVRshort,

0.10

AGGAATGCAAAGARCAG

407

P. ovale

VIRshort,

0.10

AAGGACTTCCAAGCC

476

P. vivax

NG-PCR‡

NewPLFshort

0.50

CTATCAGCTTTTGATGTTAG

53°C

735–773¶

Plasmodium

 

NewRevshort

0.50

CCTTAACTTTCGTTCTTG

  

Plasmodium

  1. *First PCR performed at final volume of 50 μl and 5 μl DNA template.
  2. †Second multiplex PCR performed at final volume of 25 μl with 2 μl template from first PCR product.
  3. ‡Second genus PCR performed at final volume of 50 μl with 2 μl template from first PCR amplification.
  4. §Size depending on Plasmodium species: P. malariae 821 bp, P. falciparum 787 bp, P. vivax 783 bp, P. ovale 794 bp, P. knowlesi 807 bp, P. cynomolgi 785 bp.
  5. ¶Size depending on Plasmodium species: P. malariae 773 bp, P. falciparum 739 bp, P. vivax 735 bp, P. ovale 746 bp, P. knowlesi 763 bp. P. cynomolgi 738 bp.