Background
Methods
Samples
Human sample | Year of Collection | Origin | |
23PA | 1996 | Peixoto de Azevedo, MT, Amazon Region, BR | |
50PA | 1996 | Peixoto de Azevedo, MT, Amazon Region, BR | |
66PA | 1996 | Peixoto de Azevedo, MT, Amazon Region, BR | |
A | 2000 | Iguape, SP, Atlantic Forest, BR | |
I11 | 2002 | Iporanga, SP, Atlantic Forest, BR | |
58 | 2004 | São Paulo, SP (Blood transfusion from Patient 72) | |
72 | 2004 | Juquitiba, SP, Atlantic Forest, BR | |
23a | 2005 | Juquitiba, SP, Atlantic Forest, BR | |
190 | 2006 | Juquitiba, SP, Atlantic Forest, BR | |
157a | 2007 | Juquitiba, SP, Atlantic Forest, BR | |
222a | 2007 | Juquitiba, SP, Atlantic Forest, BR | |
Sample | Year of Collection | Source | Origin |
PIII | 1987 |
Saimiri sciureus
| Iquitos, Peru |
M95 | 2010 |
Anopheles (K.) cruzii
| Itanhaém, SP, Atlantic Forest, BR |
P169 | 2010 |
Callicebus dubius
| Porto Velho, RO, Amazon Region, BR |
P171 | 2010 |
Pithecia sp.
| Porto Velho, RO, Amazon Region, BR |
P177 | 2011 |
Callicebus caligatus
| Manaus, AM, Amazon Region, BR |
P182 | 2011 |
Callicebus caligatus
| Manaus, AM, Amazon Region, BR |
Design of PmMSP1-specific primers
GenBank accession no. | |||
---|---|---|---|
Plasmodium species (strain) | Host species | Aminoacid | Nucleotide |
berghei
|
Grammomys surdaster
| AAF13063.1 | AF187232.1 |
chabaudi
|
Thamnonys rutilans
| AAA29499.1 | L22982.1 |
coatneyi
|
Macaca fascicularis
| BAF74048.1 | AB266180.1 |
cynomolgi
|
Macaca sinica
| BAF74063.1 | AB266195.1 |
falciparum (MAD20) |
Homo sapiens
| A26868 | X05624.2 |
falciparum (K1) |
Homo sapiens
| CAA27070.1 | X03371.1 |
fragile
|
Macaca sinica
| BAF74049.1 | AB266181.1 |
gallinaceum
|
Gallus gallus
| CAH10838.1 | AJ809338.1 |
hylobati
|
Hylobati moloch
| BAF74050.1 | AB266182.1 |
inui
|
Cynopithecus niger
| BAF74051.1 | AB266183.1 |
knowlesi
|
Macaca fascicularis
| BAF74052.1 | AB266184.1 |
malariae (MM1A) |
Homo sapiens
| ACZ51237.1 | FJ824669 |
ovale (OM1A) |
Homo sapiens
| ACZ51238.1 | FJ824670 |
ovale (OM1B) |
Homo sapiens
| ACZ51239.1 | FJ824671 |
reichenowi
|
Pan troglodytes
| CAH10285.1 | AJ786604.1 |
simiovale
|
Macaca sinica
| BAF74053.1 | AB266185.1 |
vivax (Belem) |
Homo sapiens
| AAN86208.1 | AF435594.1 |
vivax (Sal1) |
Homo sapiens
| EDL45115.1 | AAKM01000007.1 |
yoelii
|
Thamnonys rutilans
| EAA17822.1 | AABL01001865.1 |
Region | PCR primers | Amplification conditions | Amplicon sizea |
---|---|---|---|
Fragment 1
| |||
First Reaction | F:5′-GCA TCT AAA TGG CTA CTG TGA TAT AC-3′ | 94 °C 5 min (1×); 94 °C 1 min, 53 °C 2 min, 72 °C 2 min (25×); 72 °C 5 min (1×) | 595 bp |
R:5′-GGC ATC TGT AAA TAG ACC ATC C-3′ | |||
Nested Reaction | F:5′-TAA TGA AAA GGA ATT AGA AAT G-3′ | 94 °C 5 min (1×); 94 °C 1 min, 43 °C 2 min, 72 °C 2 min (30×); 72 °C 5 min (1×) | 533 bp |
R:5′-CCA TAT TGA ATA CTA TAT TTT TC-3′ | |||
Fragment 2
| |||
First Reaction | F: 5′-CCA TAC TAT TTA ATT GCA CTA AAG-3′ | 94 °C 5 min (1×); 94 °C 1 min, 58 °C 2 min, 72 °C 2 min (25×); 72 °C 5 min (1×) | 723 bp |
R:5′-ACA CAC ATA AGC AGT TTT CAA AAA G-3′ | |||
Nested Reaction | F:5′-AGG GAA ATT GAT AAA TTA AAT ATT TC-3′ | 94 °C 5 min (1×); 94 °C 1 min, 58 °C 2 min, 72 °C 2 min (30×); 72 °C 5 min (1×) | 666 bp |
R:5′-CTT TTC AAG ATA TTG CAA TTT GGA-3′ | |||
Fragment 3
| |||
First Reaction | F:5′-GCA ATG TTC TAC AAA ATC AGT ACA AAG-3′ | 94 °C 5 min (1×); 94 °C 1 min, 53 °C 2 min, 72 °C 2 min (25×); 72 °C 5 min (1×) | 765 bp |
R:5′-CTA CAA AAG CTG CTA GTA CAT GTC T-3′ | |||
Nested Reaction | F:5′-GAA GAA GAA TGA CAA ACT TAA AAA C-3′ | 94 °C 5 min (1×); 94 °C 1 min, 43 °C 2 min, 72 °C 2 min (30×); 72 °C 5 min (1×) | 632 bp |
R:5′-CTT TTA AAA TAC TAT ATT CTT TAA TAT G-3′ | |||
Fragment 4
| |||
First Reaction | F:5′-GAG GTG ATC GTG TTT CCC ATT G-3′ | 94 °C 5 min (1×); 94 °C 1 min, 48 °C 2 min, 72 °C 2 min (25×); 72 °C 5 min (1×) | 532 bp |
R:5′-CAT TTT CAG TTT TTT TAT CAC CC-3′ | |||
Nested Reaction | F:5′-GTA AGA AAG AAA AGG AAA ATC CAT TAG-3′ | 94 °C 5 min (1×); 94 °C 1 min, 49 °C 2 min, 72 °C 2 min (30×); 72 °C 5 min (1×) | 443 bp |
R:5′-CTG AGT CTT GTA CAA CTT GGT C-3′ | |||
Fragment 5
| |||
First Reaction | F:5′-GAC CAA GTT GTA CAA GAC TCA G-3′ | 94 °C 5 min (1×); 94 °C 1 min, 48 °C 2 min, 72 °C 2 min (25×); 72 °C 5 min (1×) | 737 bp |
R:5′-GTA AGT TAA ACA TAA TTA ATA AAG CTG-3′ | |||
Nested Reaction | F: 5′-GGG TGA TAA AAA AAC TGA AAA TG-3′ | 94 °C 5 min (1×); 94 °C 1 min, 49 °C 2 min, 72 °C 2 min (30×); 72 °C 5 min (1×) | 631 bp |
R:5′-GCC GAG GAA ACT TGA AGA AC-3′ |