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Table 9 Real-time PCR results of 14-plex assay

From: Comparison of the performance of an amplicon sequencing assay based on Oxford Nanopore technology to real-time PCR assays for detecting bacterial biodefense pathogens

Organism

Strain

Agent

F. tularensis assays

Y. pestis assays

Burkholderia assays

B. anthracis assays

F. tularensis

239

A

30.63

32.97

TN

TN

240

B

26.05

TN

28.75

TN

241

C

29.80

TN

TN

30.97

Y. pestis

113

D

24.86

23.04

TN

30.28

114

E

27.75

TN

29.71

31.04

B. mallei

164

F

32.81

TN

TN

B. pseudomallei

197

G

TN

32.60

32.14

B. anthracis

708

H

27.07

28.45

28.97

PCR Assay Number

23

28

29

30

9

11

14

15

49

50

65

01

04

07

Probe Dye Channel

FAM

VIC

NED

CY5

FAM

VIC

NED

CY5

FAM

VIC

NED

FAM

VIC

NED

  1. Real-time PCR results using a mixed assay of all 14 sets of primers and probes tested on individual agents. Each sample contained a single agent, extracted in singlet and analyzed by PCR. Agent concentration are all at 2.5E+ 05 CFU/mL. Each PCR reaction employed all 14 primer/probe sets. Data shows that only agent specific primer/probe sets detected with no FP or FN. Values in the cells indicate the Ct value of an observed positive result (Ct < 40) where a positive result was expected. A minus sign (−) indicates the assay-organism combination was not tested. Cells containing TN indicate an observed negative or undetected result (Ct ≥ 40) where a positive result was not expected