Comparative Evaluation of Four Real-Time PCR Methods for the Quantitative Detection of Epstein-Barr Virus from Whole Blood Specimens Daelynn Buelow,

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Comparative Evaluation of Four Real-Time PCR Methods for the Quantitative Detection of Epstein-Barr Virus from Whole Blood Specimens Daelynn Buelow, Yilun Sun, Li Tang, Zhengming Gu, Stanley Pounds, Randall Hayden The Journal of Molecular Diagnostics Volume 18, Issue 4, Pages 527-534 (July 2016) DOI: 10.1016/j.jmoldx.2016.02.002 Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Linearity, limits of detection, and limits of quantification for type 1 by each method. Linearity of artificially seeded whole blood samples with type 1 as measured by real-time PCR. A total of nine replicates of each concentration were tested, and all replicates were plotted against concentrations determined by droplet digital PCR. Ct, cycle threshold; LDT, laboratory-developed test. The Journal of Molecular Diagnostics 2016 18, 527-534DOI: (10.1016/j.jmoldx.2016.02.002) Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 Colinear regression of type 1 and type 2 in artificially seeded whole blood samples. Epstein-Barr virus type 1 and type 2 were both serial diluted, extracted, and used to generate the linear regression lines. Ct was plotted against the droplet digital PCR concentrations. Graphs represent the three analyte-specific reagents (A–C) and one LDT (D) evaluated. Ct, cycle threshold; LDT, laboratory-developed test. The Journal of Molecular Diagnostics 2016 18, 527-534DOI: (10.1016/j.jmoldx.2016.02.002) Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 3 Quantitative pairwise correlation of patient samples between the different methods. Values are represented in log10 copy/mL. Intercept, slope, and R2 for each comparison are indicated within each graph, with numbers in parentheses indicating the 95% CI. Comparison between Luminex and LDT (A), Luminex and Focus (B), Luminex and ELITech (C), ELITech and LDT (D), ELITech and Focus (E), and Focus and LDT (F). LDT, laboratory-developed test. The Journal of Molecular Diagnostics 2016 18, 527-534DOI: (10.1016/j.jmoldx.2016.02.002) Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions