Volume 21, Number 10—October 2015
Dispatch
Detection of Mixed Infections with Plasmodium spp. by PCR, India, 2014
Table 2
Characteristics of PCR primers specific for 18S rRNA gene of Plasmodium spp., India*
Genus or species |
Primer |
Sequence, 5′→3′ |
PCR product, bp |
PCR Program | No. cycles |
|||||
---|---|---|---|---|---|---|---|---|---|---|
Denaturation |
Annealing |
Elongation |
||||||||
Temp, °C |
Time, min |
Temp, °C |
Time, min |
Temp ,°C |
Time, min |
|||||
Plasmodium | F | TTAAAATTGTTGCAGTTAAAACG | 1,200 | 94 | 1 | 58 | 2 | 72 | 2 | 25 |
R | CCTGTTGTTGCCTTAAACTTC | 1,200 | 94 | 1 | 58 | 2 | 72 | 2 | 25 | |
P. falciparum | F | TTAAACTGGTTTGGGAAAACCAAATATATT | 205 | 94 | 1 | 58 | 2 | 72 | 2 | 30 |
R | ACACAATGAACTCAATCATGACTACCCGTC | 205 | 94 | 1 | 58 | 2 | 72 | 2 | 30 | |
P. vivax | F | CGCTTCTAGCTTAATCCACATAACTGATAC | 120 | 94 | 1 | 58 | 2 | 72 | 2 | 30 |
R | ACTTCCAAGCCGAAGCAAAGAAAG TCCTTA | 120 | 94 | 1 | 58 | 2 | 72 | 2 | 30 | |
P. malariae | F | ATAACATAGTTGTACGTTAAGAATAACCGC | 144 | 94 | 1 | 58 | 2 | 72 | 2 | 30 |
R | AAAATTCCCATGCATAAAAAATTATACAAA | 144 | 94 | 1 | 58 | 2 | 72 | 2 | 30 | |
P. ovale | F | ATCTCTTTTGCTATTTTTTAGTATTGGAGA | 800 | 94 | 1 | 58 | 2 | 72 | 2 | 30 |
R | GGAAAAGGACACATTAATTGTATCCTAGTG | 800 | 94 | 1 | 58 | 2 | 72 | 2 | 30 | |
P. knowlesi | F | CAGAGATCCGTTCTCATGATTTCCATGG | 209 | 95 | 0.5 | 57 | 0.5 | 72 | 0.75 | 35 |
R | CTRAACACCTCATGTCGTGGTAG | 209 | 95 | 0.5 | 57 | 0.5 | 72 | 0.75 | 35 | |
P. ovale | F | CTACTTGACATTTCTACTTACA | 938 | 95 | 1 | 50 | 1 | 72 | 1 | 35 |
R | CGTTCTTGATTAATGGAAGTAT | 938 | 95 | 1 | 50 | 1 | 72 | 1 | 35 | |
P. ovale (curtisi and wallikeri) | F | GCTGTAGCTAATACTTGCTTTA | 827 | 95 | 1 | 55 | 1 | 72 | 1 | 25 |
R | TTCACCTCTGACATCTGAATC | 827 | 95 | 1 | 55 | 1 | 72 | 1 | 25 |
*F, forward; R, reverse.