Table 2.
Primer pair sequence, concentration and amplicon size and Tm value for each SYBR®Green qPCR assay
| Targeted genus | Targeted species | Targeted gene | Primer pair name | Primer name | Primer sequence, 5′→3′ | Primer concentration (nM) | Product size (bp) | T m amplicon | Reference |
|---|---|---|---|---|---|---|---|---|---|
| Listeria | All (except L. grayi) | Invasion-associated protein (iap) | iap-50-deg | iap-31-deg-F | CAYCCGCWAGCACWGTAGTAGT | 1,000 | 78 | 75.5–76 °C L. monocytogenes, L. seeligeri, L. welshimeri and L. ivanovii; 77 °C L. innocua | This study |
| iap-50-deg-R | GCGTCRACAGTWGTSCCHTT | 1,000 | |||||||
| Phosphoribosyl pyrophosphate synthetase (prs) | prs-2-deg | prs-2-F | ATTTTCTCGCTAAATTCTAATCGTG | 250 | 60 | 71.5 °C | This study | ||
| prs-2-R-deg | CAATACCWACTTCTTTCGCAATCTC | 250 | |||||||
| Listeria | monocytogenes | Listeriolysin O (hlyA) | hlyA-177 | hlyA-177-F | TGCAAGTCCTAAGACGCCA | 250 | 112 | 74 °C | Nogva et al. (2000) |
| hlyA-177-R | CACTGCATCTCCGTGGTATACTAA | 250 | |||||||
| hlyA-146-deg-tronc | hlyA-146-deg-tronc-F | AAATCTGTCTCAGGYGATGT | 1,000 | 103 | 73.5–74 °C for L. monocytogenes 1/2a, 1/2c, 3a, 3c, 4a, 4d and 4e and 75 °C for L. monocytogenes 1/2b, 3b, 4b and 4c | Adapted from Hough et al. (2002) | |||
| hlyA-146-R | CGATGATTTGAACTTCATCTTTTGC | 1,000 |
In italics are degenerated nucleotides: R = A or G, Y = C or T, W= A or T, S = C or G, H = A or C or T