Table 2.
Specificity and multiplexing of the PRI-lock probe system in conventional real-time PCR and in the Biotrove OpenArray™ for single and multiple targets.
| Single target | Multiplex mix 1 | Multiplex mix 2 | Multiplex mix 3 | |||||
| PRI-lock Probe | CT(AB) | CT(BT) | CT(AB) | CT(BT) | CT(AB) | CT(BT) | CT(AB) | CT(BT) |
| Phytophthora spp. | 15.5 (0.14) | 16.4 (0.17) | 15.7 (0.10) | 16.7 (0.25) | 16.1 (0.04) | 16.8 (0.12) | 16.3 (0.16) | 15.9 (0.09) |
| P. infestans | 14.8 (0.12) | 15.4 (0.18) | -- | -- | 15.2 (0.08) | 15.7 (0.12) | 15.1 (0.02) | 15.0 (0.08) |
| A. tumefaciens | 16.4 (0.24) | 16.2 (0.04) | -- | -- | -- | -- | 15.8 (0.28) | 15.9 (0.06) |
| G. Proteo bacterial spp | 17.0 (0.11) | 17.7 (0.08) | -- | -- | -- | -- | 17.7 (1.03) | 17.1 (0.04) |
| M. roridum | 16.2 (0.07) | -- | 16.4 (0.02) | 17.2 (0.05) | -- | -- | 16.7 (0.12) | 17.0 (0.16) |
| F. oxysporum | 17.4 (0.11) | -- | -- | -- | -- | -- | 17.2 (0.08) | 17.3 (0.04) |
| E. carotovora carotovora | 15.1 (0.15) | -- | -- | -- | -- | -- | 15.2 (0.24) | 15.4 (0.05) |
| V. dahliae | 16.9 (0.01) | -- | -- | -- | -- | -- | 16.1 (0.04) | 15.8 (0.03) |
| V. alboatrum/V. tricorpus | 19.6 (0.11) | -- | -- | -- | -- | -- | 19.9 (0.29) | 18.0 (0.10) |
| M. hapla | 17.0 (0.30) | -- | -- | -- | -- | -- | 18.2 (0.23) | 16.9 (0.11) |
| R. solani AG 2-2 | 16.3 (0.03) | -- | -- | -- | 15.8 (0.10) | 17.1 (0.12) | 16.2 (0.03) | 16.2 (0.04) |
| R. solani AG 4-1 | 14.0 (0.02) | -- | -- | -- | 14.6 (0.13) | 15.0 (0.11) | 14.5 (0.13) | 14.4 (0.09) |
| R. solani AG 4-2 | 19.3 (0.10) | -- | -- | -- | 19.5 (0.14) | 20.0 (0.18) | 19.3 (0.07) | 18.8 (0.03) |
The PRI-lock probes were ligated on 106 targets/μl ligation mixture. CT values were normalized using the ILC control PRI-lock probe. Data represent average CT values of three PCR replicates in the conventional real-time platform (n = 3) and of four PCR replicates in the Biotrove OpenArray™ (n = 4). Standard deviations are indicated between brackets. AB: samples run on the conventional real-time PCR platform. BT: Samples tested on the Biotrove OpenArray™ platform. Multiplex mix 1: P. soj. and M. ror. DNA. Multiplex mix 2: P. inf., R. sol. AG 2-2, R. sol. AG 4-1 and R. sol. AG 4-2 DNA. Multiplex mix 3: ligation mixture containing all the DNA targets indicated in table 2. --: not tested.