Table 1.
Oligonucleotide primer sequences and PCR reaction conditions for species identification
| Primer for species ID | Sequence 5’ ⇒ 3’ | PCR product size (bp) | PCR conditions |
|---|---|---|---|
|
Anopheles gambiae
s.l. species | |||
| Universal primer |
GTGTGCCCCTTCCTCGATGT |
468 |
93°C 3’ followed by 35 cycles (93°C 30”; 50°C 30”; 72°C 1’); 93°C 30”; 50°C 30”; 72°C 10’ |
|
Anopheles gambiae s.s. |
CTGGTTTGGTCGGCACGTTT |
390 |
|
|
Anopheles merus/melax |
TGACCAACCCACTCCCTTGA |
464 |
|
|
Anopheles arabiensis |
AAGTGTCCTTCTCCATCCTA |
315 |
|
|
Anopheles quadrianulatus |
CAGACCAAGATGGTTAGTAT |
153 |
|
|
Anopheles funestus
s.l. species | |||
| Universal primer |
TGTGAACTGCAGGACACAT |
|
30 cycles (94°C 30”; 40°C 30”; 72°C 30”); 72°C 10’ |
|
Anopheles funestus s.s. |
GCATCGATGGGTTAATCATG |
460 |
|
|
Anopheles vaneedeni |
TGTCGACTTGGTAGCCGAAC |
555 |
|
|
Anopheles rivulorum |
CAAGCCGTTCGACCCTGATT |
400 |
|
|
Anopheles parensis |
TGCGGTCCCAAGCTAGGTTC |
235 |
|
|
Anopheles leesoni |
TACACGGGCGCCATGTAGTT |
146 |
|
|
Wuchereria bancrofti
| |||
|
NV-1 |
CGTGATGGCATCAAAGTAGCG |
188 |
94°C 3’; followed by 35 cycles (94°C 1’; 55°C 1’; 72°C 2’); 94°C 1’; 55°C 1’; 72°C 10” |
| NV-2 | CCCTCACTTACCATAAGACAAC | 188 | |