Table 1

Specificity and sensitivity of the novel primers in the detection of Las as shown by qRT-PCR assay
Primer pairs Target gene Las CT value of the qRT-PCR#
Negative control Other controls
CT value R2 value Slope Laf Lam Healthy plant tissue Water C1 C2 C3 C4 C5 C6
P1 CLIBASIA_05555 20.54 0.9944 -0.2883 UD UD UD UD UD UD UD UD UD UD
P2 CLIBASIA_04315 19.99 0.9867 -0.2849 UD UD UD UD UD UD UD UD UD UD
P3 CLIBASIA_05575 20.15 0.9991 -0.2847 UD UD UD UD UD UD UD UD UD UD
P4 CLIBASIA_05465 19.52 0.9618 -0.2897 UD UD UD UD UD UD UD UD UD UD
P5 CLIBASIA_01460 19.48 0.9995 -0.2969 UD UD UD UD UD UD UD UD UD UD
P6 CLIBASIA_05145 22.29 0.9971 -0.3057 UD UD UD UD UD UD UD UD UD UD
P7 CLIBASIA_05545 20.11 0.9972 -0.3407 UD UD UD UD UD UD UD UD UD UD
P8 CLIBASIA_05560 19.92 0.9982 -0.3132 UD UD UD UD UD UD UD UD UD UD
P9 CLIBASIA_02025 20.12 0.9875 -0.2743 UD UD UD UD UD UD UD UD UD UD
P10 CLIBASIA_05605 20.18 0.9945 -0.2781 UD UD UD UD UD UD UD UD UD UD
P11 CLIBASIA_03090 23.61 0.9997 -0.2867 UD UD UD UD UD UD UD UD UD UD
P12 CLIBASIA_03875 27.47 0.9992 -0.2563 UD UD UD UD UD UD UD UD UD UD
P13 CLIBASIA_02305 UD NT NT UD UD UD UD UD UD UD UD UD UD
P14 CLIBASIA_05495 21.25 0.9974 -0.2594 UD UD UD UD UD UD UD UD UD UD
P15 CLIBASIA_02660 UD NT NT UD UD UD UD UD UD UD UD UD UD
P16 CLIBASIA_02715 20.26 0.9411 -0.3480 UD UD UD UD UD UD UD UD UD UD
P17 CLIBASIA_03110 20.11 0.9994 -0.2786 UD UD UD UD UD UD UD UD UD UD
P18 CLIBASIA_03675 20.02 0.9967 -0.2780 UD UD UD UD UD UD UD UD UD UD
P19 CLIBASIA_03725 19.91 NT NT 35.29 UD UD UD UD UD UD UD UD UD
P20 CLIBASIA_03955 21.08 NT NT UD UD UD UD 37.41 UD UD UD UD UD
P21 CLIBASIA_04030 20.30 NT NT UD UD UD UD 32.93 UD UD UD UD UD
P22 CLIBASIA_04150 24.00 NT NT UD UD UD UD UD UD UD UD UD UD
P23 CLIBASIA_04310 20.76 0.991 -0.2976 UD UD UD UD UD UD UD UD UD UD
P24 CLIBASIA_04330 20.85 0.9986 -0.2635 UD UD UD UD UD UD UD UD UD UD
P25 CLIBASIA_04405 21.60 0.9987 -0.3051 UD UD UD UD UD UD UD UD UD UD
P26 CLIBASIA_04425 20.41 0.9994 -0.3032 UD UD UD UD UD UD UD UD UD UD
P27 CLIBASIA_02645 21.77 NT NT 38.61 UD UD UD UD UD UD UD UD UD
P28 CLIBASIA_04515 22.00 NT NT 38.63 UD UD UD UD UD UD UD UD UD
P29 CLIBASIA_04530 19.00 0.9919 -0.2852 UD UD UD UD UD UD UD UD UD UD
P30 CLIBASIA_04550 22.48 0.9938 -0.2708 UD UD UD UD UD UD UD UD UD UD
P31 CLIBASIA_05230 21.68 0.9941 -0.2771 UD UD UD UD UD UD UD UD UD UD
P32 CLIBASIA_05480 21.48 0.988 -0.2776 UD UD UD UD UD UD UD UD UD UD
P33 CLIBASIA_04475 20.84 0.9913 -0.2644 UD UD UD UD NT UD UD UD UD UD
P34 CLIBASIA_05505 22.70 0.9893 -0.2791 UD UD UD UD NT UD UD UD UD UD
CQULA04F/R β-operon 22.11 NT NT UD UD UD UD NT NT NT NT NT NT
LJ900f/r Prophage 22.25 NT NT UD UD UD UD NT NT NT NT NT NT
HLBas/r 16Sas 24.33 0.9998 -0.3057 NT NT UD UD NT NT NT NT NT NT
HLBam/r 16Sam NT NT NT NT 24.68 UD UD NT NT NT NT NT NT
HLBaf/r 16Saf NT NT NT 21.28 NT UD UD NT NT NT NT NT NT
COXf/r Cox 14.80 NT NT 15.21 18.54 16.15 UD NT NT NT NT NT NT

Las-infected psyllids total DNA was serially diluted spanning up to five logs and used as a template in the qRT-PCR assay. R2 and slope were further calculated from a plot of CT values versus log dilution factor. #qRT-PCR was conducted by using template DNA samples of Las, Laf, Lam, C1: Colletotrichum acutatum KLA-207, C2: Elsinoe fawcettii, C3: Xanthomonas axonopodis pv. citrumelo1381, C4: Xanthomonas citri subsp. citri Aw, C5: Xanthomonas citri subsp. citri A*, C6: Xanthomonas citri subsp. citri 306. The CT values are average of three replicates for each primer pair. UD: undetected; NT: Not tested.

Kogenaru et al.

Kogenaru et al. BMC Microbiology 2014 14:39   doi:10.1186/1471-2180-14-39

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