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Figure 2 | Malaria Journal

Figure 2

From: First case of Plasmodium knowlesi infection in a Japanese traveller returning from Malaysia

Figure 2

Nested PCR results from the inner primers. The region amplified was a partial sequence of the cytochrome b gene (mitochondrial DNA) from different malaria parasite species (~130-180 bp). M: DNA size marker, P: patient, Pf: Plasmodium falciparum, Pv: Plasmodium vivax, Po: Plasmodium ovale, Pm: Plasmodium malariae and, Pk: Plasmodium knowlesi. PC: positive control, NC: negative control. Forward primers for the outer PCR and the inner PCR were the same as those reported by Putaporntip et al. [15]. The sequences of the reverse primers used for the outer PCR and the inner PCR are as follows: PCBR-ed (5′-ACATAATTATAACCTTACGGTCTG-3′), PfCBR-ed (5′-GATTTGTTCCGCTCAATAC-3′), PvCBR-ed (5′-CTGTATTGTTCTGCTCAA-3′), PoCBR-ed (5′-CTGTATTGTTCTGCTCAT-3′), PmCBR-ed (5′-CTGTATTGTTCTGCACAG-3′), PkCBR-ed (5′-GTATTGTTCTAATCAGTGTA-3′). Nested PCR primers for the small-subunit rRNA from parasite nuclear DNA were the same as those reported by Kimura et al. [16] (gel not shown). The inner PCR primer (reverse) used to amplify P. knowlesi DNA (SS-rRNA-Pk-R) had the following sequence: 5′-AAGAGTTCTAATCTCCGGAGAGAAAAG-3′. DNA sequences of the partial cytochrome b gene and SSU rRNA gene were deposited in the DNA Data Bank of Japan (DDBJ). The DDBJ accession numbers of the DNA sequences of the partial cytochrome b gene and SSU rRNA gene were AB787188 and AB787187, respectively. A positive band for the patient′s blood sample with P. knowlesi primer sets was shown.

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