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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.