Background
Methods
Database accession of strains and isolates used in this study
Blood sample collection
DNA preparation from blood samples
PCR amplification of PvMCA1 gene and Pvmdr1 gene
Genes | Primers | Sequences | PCR program |
---|---|---|---|
PvMCA1-cd | MCA1-F1 | 5′-CATGGAAACAAAAAAAAGG-3′ | 95 °C 10′, 35 cycles 94 °C 30″ 56 °C 30″ 72 °C 2′ |
MCA1-R1 | 5′-CGAAAACTCCATATCTTTGC-3′ | ||
pvmdr1
| mdr1-F1 | 5′-ATAGTCATGCCCCAGGATTG-3′ | 95 °C 10′, 45 cycles 95 °C 10″, 60 °C 10″ and 72 °C 30″ |
mdr1-R1 | 5′-ACGTTTGGTCTGGACAAGTAT-3′ | ||
Pvcsp
| VCSP-OF | 5′-ATGTAGATCTGTCCAAGGCCATAAA-3′ | Primary reaction |
VCSP-OR | 5′-TAATTGAATAATGCTAGGACTAACAATATG-3′ | 95 °C 10′, 40 cycles 95 °C 1′, 58 °C 2′ and 72 °C 2′ | |
VCSP-NF | 5′-GCAGAACCAAAAAATCCACGTGAAAATAAG-3′ | Nested reaction | |
VCSP-NR | 5′-CCAACGGTAGCTCTAACTTTATCTAGGTAT-3′ | 30 cycles 95 °C 1′, 62 °C 2′ and 72 °C 2′ |
Pvcsp genotyping of P. vivax isolates
Results
Analysis of pvmdr1 gene polymorphisms
Location/ID-isolate | M958Y | Y976F/Y976V | F1076L/F1076I/F1076T |
---|---|---|---|
NCBI-AY618622a
| M | Y | F |
MAURIT-61606 | M | Y | F |
MAURIT-00721 | M | Y |
L
|
SUDAN-KH-179 | M | Y |
L
|
SUDAN-KH-193 | M | Y |
L
|
OMAN-8 | M | Y |
L
|
OMAN-39 | M | Y |
L
|
OMAN-3 | M | Y |
L
|
OMAN-51 | M | Y |
L
|
OMAN-48 | M | Y |
L
|
SUDAN-KH-144 | M |
F
|
L
|
SUDAN-KH-142 | M |
F
|
L
|
SUDAN-KH047 | M |
F
|
L
|
SUDAN-KH-145 |
Y
|
F
|
L
|
MAURIT-06727 |
Y
|
V
|
I
|
MAURIT-Mou15 |
Y
|
V
|
T
|
MAURIT-06826 |
Y
|
V
|
T
|
MAURIT-Chi43 |
Y
|
V
|
T
|
MAURIT-Well30 |
Y
|
V
|
T
|
MAURIT-94241 |
Y
|
V
|
T
|
MAURIT-Jid34 |
Y
|
V
|
T
|
MAURIT-63933 |
Y
|
V
|
T
|
SUDAN-KH-130 |
Y
|
V
|
T
|
SUDAN-KH-173 |
Y
|
V
|
T
|
SUDAN-KH-103 |
Y
|
V
|
T
|
SUDAN-KH-195 |
Y
|
V
|
T
|
OMAN-52 |
Y
|
V
|
T
|
OMAN-36 |
Y
|
V
|
T
|
OMAN-13 |
Y
|
V
|
T
|
OMAN-17 |
Y
|
V
|
T
|
OMAN-47 |
Y
|
V
|
T
|
Comparison of P. vivax caspase-like sequences of reference strains from different areas
Analysis of PvMCA1 catalytic domain
Location/ID-isolate | H282M/H282K | C305T/C305R | H372D/H372T | C428R |
pvmdr1
|
---|---|---|---|---|---|
PlasmoDB-PVX_114725a
| H | C | H | C | MYF |
MAURIT-00721 | H | C | H | C | MYL
|
SUDAN-KH-142 | H | C | H | C | MFL
|
SUDAN-KH-193 | H | C | H | C | MYL
|
OMAN-48 | H | C | H | C | MYL
|
MAURIT-06727 | H |
T
|
D
|
R
|
YVI
|
MAURIT-Jid34 | H |
T
|
D
|
R
|
YVT
|
MAURIT-63933 | H |
T
|
D
|
R
|
YVT
|
SUDAN-KH-195 | H |
T
|
D
|
R
|
YVT
|
OMAN-36 | H |
T
|
D
|
R
|
YVT
|
OMAN-13 | H |
T
|
D
|
R
|
YVT
|
OMAN-17 | H |
T
|
D
|
R
|
YVT
|
OMAN-51 | ND |
T
|
D
|
R
| MYL
|
SUDAN-KH-103 | ND |
T
|
D
|
R
|
YVT
|
MAURIT-94241 |
M
|
T
|
D
|
R
|
YVT
|
MAURIT-Well30 |
M
|
T
|
D
|
R
|
YVT
|
MAURIT-Mou15 |
K
|
R
|
T
|
R
|
YVT
|
MAURIT-06826 |
K
|
R
|
T
|
R
|
YVT
|
MAURIT-61606 |
K
|
R
|
T
|
R
| MYF |
MAURIT-Chi43 |
K
|
R
|
T
|
R
|
YVT
|
SUDAN-KH-179 |
K
|
R
|
T
|
R
| MYL
|
SUDAN-KH-144 |
K
|
R
|
T
|
R
| MFL
|
SUDAN-KH-145 |
K
|
R
|
T
|
R
|
YFL
|
SUDAN-KH-130 |
K
|
R
|
T
|
R
|
YVT
|
SUDAN-KH-173 |
K
|
R
|
T
|
R
|
YVT
|
OMAN-52 |
K
|
R
|
T
|
R
|
YVT
|
OMAN-8 |
K
|
R
|
T
|
R
| MYL
|
OMAN-39 |
K
|
R
|
T
|
R
| MYL
|
OMAN-3 |
K
|
R
|
T
|
R
| MYL
|