Background
Methods
Clinical isolates
Drugs
Maturation of P. falciparumisolates
In vitrodrug sensitivity assay
Genetic polymorphisms in pfmdr1, pfcrt and pfATPase 6
Genes, Codons | Primer names | Primers | T°C | Restriction enzyme | sizes (bp)* |
---|---|---|---|---|---|
Pfmdr1, N86Y
| mdr86D1 | TTTACCGTTTAAATGTTTACCTGC |
45
|
Afl III
|
126 +165
|
mdr86D2 | CCATCTTGATAAAAAACACTTCTT | ||||
Pfmdr1, D1246Y,
| mdr1246D1 | AATGTAAATGAATTTTCAAACC |
45
|
Bgl II
|
113 + 90
|
mdr1246D2 | CATCTTCTCTTCCAAATTTGATA | ||||
ATPase 6, G110A
| ATP6-110F | CGTTGAACTTATTATATCTTTGTC |
60
|
Mbo II
|
103+94+92+40
|
ATP6-110R | TTTCATATCTAATAAAGTTAACACG | ||||
ATPase 6, A2694T
| ATP6-2694F | GAATTGTTTTCTGTAGAACTGAAC |
55
|
Tas I
|
142 + 39
|
ATP6-2694R | ATCTGATGCTTCTTTAGCTACC | ||||
Pfcrt, C72S
| CRT72MS | TTTATATTTTAAGTATTATTTATTTAAGTGGA |
55
|
Mbo I
|
55 + 38
|
76-D2 | CAAAACTATAGTTACCAATTTTG | ||||
Pfcrt, M74I
| CRT745MS | TAAGTATTATTTATTTAAGTGTATGTGTCAT |
55
|
Nla III
|
53 + 31
|
76-D2 | CAAAACTATAGTTACCAATTTTG | ||||
Pfcrt, N75M
| CRT745MS | TAAGTATTATTTATTTAAGTGTATGTGTCAT |
50
|
BspHI
|
53 + 31
|
76-D2 | CAAAACTATAGTTACCAATTTTG | ||||
Pfcrt, 76
| Pfcrt-76A | GCGCGCGCATGGCTCACGTTTAGGTGGAG |
55
|
Apo I
|
136 + 56
|
Pfcrt-76B | GGGCCCGGCGGATGTTACAAAACTATAGTTACC |
Statistical analysis
Results
In vitro drug sensitivity of P. falciparumisolates
Drug 1 | Drug 2 | Coefficient of correlation (r2) | p |
---|---|---|---|
DHA | MF | 0.73 | < 0.001 |
DHA | CQ | 0.39 | 0.006 |
DHA | MDAQ | 0.37 | 0.08 |
CQ | MF | 0.45 | 0.04 |
CQ | MDAQ | 0.34 | 0.5 |
MDAQ | MF | 0.30 | 0.08 |
Drug sensitivity and P. falciparumgene polymorphisms
Drug | Profils | N | N° (%), of strains with polymorphism : (Pfmdr) | |||||||
---|---|---|---|---|---|---|---|---|---|---|
N86Y | D1246Y | |||||||||
N | Y | MIX | p | D | Y | MIX | p | |||
CQ | Resistant (IC50>100 nM) | 20 | 13 (65.0%) | 6 (30.0%) | 1 (5.0%) | 0.39 | 14 (70.0%) | 2 (10.0%) | 4 (20.0%) |
0.3
|
Sensitive (IC50<100 nM) | 26 | 15 (57.7%) | 11 (42.3%) | 0 (0.0%) | 18 (69.2%) | 6 (23.1%) | 2 (7.7%) | |||
MDAQ | Resistant (IC50 ≥ 60 nM) | 24 | 14 (58.3%) | 9 (37.5%) | 1 (4.2%) | 0.28 | 16 (66.7%) | 3 (12.5%) | 5 (20.8%) |
0.09
|
Sensitive (IC50 < 60 nM) | 22 | 12 (54.5%) | 10 (45.5%) | 0 (0.0%) | 20 (90.9%) | 0 (0.0%) | 2 (9.1%) | |||
MF | Resistant (IC50>20 nM) | 11 | 6 (54.5%) | 5 (45.5%) | 0 (0.0%) | 0.7 | 7 (63.6%) | 2 (18.2%) | 2 (18.2%) | 0.23 |
Sensitive (IC50<20 nM) | 35 | 22 (62.8%) | 12 (34.3%) | 1 (2,9%) | 28 (80.0%) | 2 (5.7%) | 5 (14.3%) | |||
DHA | Reduced susceptibility (IC50 ≥ 10 nM) | 11 | 6 (54.5%) | 5 (45.5%) | 0 (0.0%) | 0.7 | 9 (81.8%) | 1 (9.1%) | 1 (9.1%) | 0.53 |
Sensitive (IC50<10 nM) | 35 | 22 (62.8%) | 12 (34.3%) | 1 (2.9%) | 27 (77.1%) | 2 (5.7%) | 6 (17.1%) |
Drug | Profils | N | N° (%), of strains with polymorphism : (Pfcrt) | |||
---|---|---|---|---|---|---|
HAPLOTYPE 72-76 | ||||||
CVIEK | CVIET | MIX | p | |||
CQ | Resistant (IC50 ≥ 100 nM) | 20 | 0 (0.0%) | 20 (100.0%) | 0 (0.0%) |
0.56
|
Sensitive (IC50 < 100 nM) | 26 | 1 (3.8%) | 25 (96.2%) | 0 (0.0%) | ||
MDAQ | Resistant (IC50 ≥ 60 nM) |
24
|
1 (4.2%)
|
23 (95.8%)
|
0 (0.0%)
|
0.29
|
Sensitive (IC50 < 60 nM) |
22
|
0 (0.0%)
|
22 (100%)
|
0 (0.0%)
| ||
MF | Resistant (IC50 ≥ 20 nM) | 11 | 0 (0.0%) | 11 (100.0%) | 0 (0.0%) |
0.58
|
Sensitive (IC50 < 20 nM) | 35 | 1 (2.9%) | 34 (97.1%) | 0 (0.0%) | ||
DHA | Reduced susceptibility (IC50 ≥ 10 nM) | 11 | 1 (9.1%) | 10 (90.9%) | 0 (0.0%) |
0.23
|
Sensitive (IC50 < 10 nM) | 35 | 0 (0.0%) | 35 (100.0%) | 0 (0.0%) |
Drug | Profils | N | Strains with polymorphism PfATPase 6 A2694T | |||
---|---|---|---|---|---|---|
A | T | MIX | p | |||
CQ | Resistant (IC50>100 nM) | 20 | 14 (70.0%) | 5 (25.0%) | 1 (5.0%) | 0.45 |
Sensitive (IC50<100 nM) | 26 | 20 (76.9%) | 5 (19.2%) | 1 (3.8%) | ||
MDAQ | Resistant (IC50 ≥ 60 nM) | 24 | 17 (69.2%) | 6 (26.9%) | 1 (3.8%) | 0.6 |
Sensitive (IC50 < 60 nM) | 22 | 17 (80.0%) | 4 (15.0%) | 1 (5.0%) | ||
MF | Resistant (IC50>20 nM) | 11 | 7 (63.6%) | 3 (27.3%) | 1 (9.1%) | 0.4 |
Sensitive (IC50<20 nM) | 35 | 27 (77.1%) | 7 (20.0%) | 1 (2.9%) | ||
DHA | Reduced susceptibility (IC50 ≥ 10 nM) | 11 | 8 (72.7%) | 3 (27.3%) | 0 (0.0%) | 0.4 |
Sensitive (IC50<10 nM) | 35 | 26 (74.3%) | 7 (20.0%) | 2 (5.71%) |