Volume 15, Number 5—May 2009
Research
Chloroquine-Resistant Haplotype Plasmodium falciparum Parasites, Haiti
Berlin L. Londono, Thomas P. Eisele, Joseph Keating, Adam Bennett, Chandon Chattopadhyay, Gaetan Heyliger, Brian Mack, Ian Rawson, Jean-Francois Vely, Olbeg Désinor, and Donald J. Krogstad
Table 1
Primers used to amplify Plasmodium falciparum DNA during study in Haiti*
| Primers (5′ → 3′) | Amplicon, bp | Tm, °C | Reference |
|---|---|---|---|
| Primers for P. falciparum species-specific SSU rRNA gene | 276 | (22) | |
| Forward: AACAGACGGGTAGTCATGATTGAG | 56.5 | ||
| Reverse: GTATCTGATCGTCTTCACTCCC | 54.5 | ||
| Primers for single-step pfcrt gene PCR | 170 | (23) | |
| Forward: TgTgCTCATgTGTTTAAACTT | 50.6 | ||
| Reverse: AATAAAgTTgTgAgTTTCggA | 49.8 | ||
| Primers for nested (2-step) pfcrt gene | 573 | (24,25) | |
| First round of amplification | |||
| Forward (CRTP1): CCGTTAATAATAAATACACGCAG | 49.9 | ||
| Reverse (CRTP2): CGGATGTTACAAAACTATAGTTACC | 51.5 | ||
| Second round of amplification | |||
| Forward (CRTD1): TGTGCTCATGTGTTTAAACTT | 134 | 50.6 | |
| Reverse (CRTD2): CAAAACTATAGTTACCAATTTTG | 46.1 |
*Nucleotides in upper case letters were conserved in 100% of sequences at those positions, and nucleotides in lower case letters were conserved in most (e.g., >50%) sequences at those positions. Tm, melting (annealing) temperature; SSU, small subunit; pfcrt, P. falciparum chloroquine resistance transporter.
