Table 1.
Primer and probe sequences for real-time quantitative assays and assay specificities.
| Assay and target locus | Primer and Probe sequences (5′–3′) | Amplicon size | Genbank accession no. | Assay specificity |
|---|---|---|---|---|
| “Generic” fungal assay | F: AGA TAC CGT CGT AGT CTT AAC CAT AAA CT; | 131 bp | B. bassiana: AY245649 | Amplifies: B. bassiana, M. anisopliae (var. |
| 18S rRNA gene | R: TTC AGC CTT GCG ACC ATA CT; | 132 bp | M. anisopliae var. acridium: AF487275 | acridium) isolate 189, M. anisopliae (var. anisopliae) isolate ICPE30 |
| Probe: 6-FAM-CGT TCG GCA CCT TAC –MGB | M. anisopliae var. anisopliae: AF487273 | Not amplify: mouse, mosquito, P. chabaudi | ||
| Beauveria bassiana assay | F: GCC GGC CCT GAA ATG G; | 121 bp | AF345539 | Amplifies: B. bassiana |
| ITSII rRNA gene | R: GAT TCG AGG TCA ACG TTC AGA AG; | Not amplify: M. anisopliae 189, M. anisopliae | ||
| Probe: 6-FAM-ACA GCT CGC ACC GGA-MGB | ICPE30, background fungi, mouse, mosquito, P. chabaudi | |||
| Metarhizium anisopliae | F: GGA TCG GCG AAG CTT TTT TCA; | 99 bp | EU307907 | Amplifies: M. anisopliae 189 |
| var. acridium (189) | R: CCC GTT GCG AGT GAG TTA CTA; | Not amplify: M. anisopliae IC30, B. bassiana, | ||
| ITSII rRNA gene | Probe: 6-FAM-CCG TCC CTT AAA TTT-MGB | background fungi, mouse, mosquito, P. chabaudi | ||
| Plasmodium chabaudi | F: TGT CAG AGG TGA AAT TCT TAG ATT TTC T; | 88 bp | DQ241815 | Amplifies: All tested P. chabaudi clones, P. |
| assay | R: ACT TTC GTT CTT GAT TAA TGG AAG TAT TT; | falciparum, P. malariae, P. berghei and P. yoelii | ||
| 18S rRNA gene | Probe: 6-FAM-CAA ACA ACT GCG AAA GC | Not amplify: mouse, mosquito, fungi (named or background) | ||