Island-wide diversity in single nucleotide polymorphisms of the Plasmodium vivax dihydrofolate reductase and dihydropteroate synthetase genes in Sri Lanka

Table 1 The sequence specific oligonucleotide probes (SSOPs) targeting SNPs/haplotypes in c57, 58, 61 and 117 of the Pvdhfr gene and c383, 553 and 585 in the Pvdhps gene and artificial positive controls.

SSOP	SSOP sequence †	Washing temperature §	Incubation time ¶
Pvdhfr		(C°)	Min.
57/58/61 FST	AC TTC AGC TCG GTG ACG A	60	12
57/58/61 FR₁T	AC TTC AGA TCG GTG ACG A	60	10
57/58/61 FR₂T	AC TTC AGG TCG GTG ACG A	60	10
57/58/61 L₁R₂M	ACCTC AGG TCG GTG ATG A	62	10
57/58/61 L₂ST	AC TTG AGC TCG GTG ACG A	60	10
57/58/61 L₂R₁T	AC TTG AGA TCG GTGACG A	64	10
57/58/61 L₃R₂T	AC TTA AGG TCG GTG ACG A	62	10
117 S	G AGA AGC AGC TGG GAG AG	60	12
117 N	G AGA AGC AAC TGG GAG AG	60	10
117 T	G AGA AGC ACC TGG GAG AG	60	10
Pvdhps
383A	A TCG TCC GCC CCT TAT GT	64	10
c383 G	A TCG TCC GGC CCT TAT GT	64	10
c553A	TC GGC CTG GGG TTTGCC A	64	10
c553G	TC GGC CTG GGG TTTGGC A	64	10
c585V	C TTT ATT GTC CAC TGC AT	64	10
Positive controls*
Pvdhfr	Sequence
PcL₃R₂T	TCCGTCGATATGAAGTACTTAAGG TCGGTGACG ACCTACGTGGATGAGTC
Pvdhps
Pc383G	GATTGACATCGGGGGGGAATCGTCCGGC CCTTATGTGGTCCCCAATCCGA
Pc553G	CTTTGATGTCGGCCTGGGGTTTGGC AAAAAGCACGACCAGTCTATTAAGC

† Sequences in bold indicate the codon in which the SNP occurs.
§ Optimal TMAC-washing temperature.
¶ Optimal incubation time for TMAC-wash
* Artificial positive controls mimicking specific mutated Pvdhfr or Pvdhps sequences.

ISSN: 1475-2875