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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.

From: Quantitative multiplex detection of plant pathogens using a novel ligation probe-based system coupled with universal, high-throughput real-time PCR on OpenArrays™

  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)
  1. 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.