Article Figures & Data
Figures
- Figure 1.
The proportion of samples with detectable levels for 67 markers on Bio-Rad (A) and 97 markers on Millipore (B) across 3 specimen types—serum (open circles), heparin plasma (filled circles), and EDTA plasma (open squares).
- Figure 2.
The CVs for 40 blinded duplicates, 20 placed within the same batch, and 20 placed across different batches. Results are shown separately for 67 Bio-Rad markers (A–C) and 97 Millipore markers (D–F) across 3 specimen types–heparin plasma, serum, and EDTA plasma.(D–F) across 3 specimen types—serum, heparin plasma, and EDTA plasma.
- Figure 3.
Spearman's correlation coefficients are shown for comparisons of the rank order of marker concentrations between T0 serum and T0 heparin plasma on Bio-Rad (A) and on Millipore (B). Comparisons between Bio-Rad and Millipore for T0 serum (C) and T0 heparin plasma (D) are also shown. T0 denotes baseline visit in the PLCO study.
- Figure 4.
Differences in ICCs between T0 heparin plasma versus T0 Serum for Bio-Rad (A) and for Millipore (B). Differences in ICCs between Millipore versus Bio-Rad for T0 serum (C) and T0 heparin plasma (D) are also shown. ICCs were estimated using variance components analyses.
Tables
- Table 1.
Summary of results for evaluation of multiplexed inflammation marker assays
Bio-Rad number of markers (%) Millipore number of markers (%) Total 67 (100.0) 97 (100.0) Markers with >25% detection Serum 56 (83.5) 89 (91.7) Heparin plasma 63 (94.0) 89 (91.7) EDTA plasma 64 (95.5) 89 (91.7) Markers with <20% CV for across-batch duplicatesa Serum 51 (76.2) 75 (77.3) Heparin plasma 52 (77.6) 69 (71.1) EDTA plasma 47 (70.1) 78 (80.4) ICCs for across-batch duplicates Serum <0.50 6 (8.9) 5 (5.1) 0.50–0.80 29 (43.3) 32 (33.0) 0.80–0.90 12 (17.9) 19 (19.5) >0.90 11 (16.4) 34 (35.1) Heparin plasma <0.50 3 (4.5) 9 (9.3) 0.50–0.80 38 (56.7) 31 (31.9) 0.80–0.90 15 (22.3) 25 (25.8) >0.90 7 (10.4) 28 (28.9) EDTA plasma <0.50 5 (7.5) 7 (7.2) 0.50–0.80 49 (73.1) 25 (25.8) 0.80–0.90 5 (7.5) 24 (24.7) >0.90 5 (7.5) 37 (38.1) Markers with acceptable performanceb 45 (67.2) 71 (73.2) -
aCVs and ICCs were calculated for 20 blinded duplicate samples for each specimen type that were placed across different batches.
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bAcceptable performance was defined as: (i) being detectable in greater than 25% of the 100 samples on all 3 specimen types and (ii) across-batch CVs of less than 20% for blinded duplicates on at least 2 of the 3 specimen types.
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- Table 2.
Summary of performance of multiplexed markers on Bio-Rad and Millipore
Bio-Rad markers with acceptable performancea Bio-Rad markers with <25% detectabilityb Millipore markers with acceptable performance Millipore markers with acceptable performancea Millipore markers with <25% detectabilityb A2M B_NGF Amylin_Total MIP_1D Eotaxin 3 CRP GM_CSF BCA_1 PP Ghrelin CTACK IFNA2 CCL19_MIP3B PYY IL_20 C Peptide IL_12P40 CCL20_MIP3A SAA IL_21 Eotaxin IL_15 CKINE SAP IL_28a Ferritin IL_1A CRP SCD30 IL_3 FGF_Basic IL_2 CTACK SCD40L IL_4 Fibrinogen IL_3 CXCL11_I_TAC SCF IL_5 Ghrelin LIF CXCL6_GCP2 SDF_1A M_CSF GIP MCP_3 CXCL9_MIG SEGFR XCL1_Lympho GLP_1 TNFB C_Peptide SGP130 Millipore markers Glucagon EGF SILRII with >20% CVc GRO Bio-Rad markers ENA_78 SIL_1RI Haptoglobin with >20% CVc Eotaxin SIL_2RA GM_CSF HGF Eotaxin_2 SIL_4R IL_10 IFNG B_NGF FGF_Basic SIL_6R IL_12P70 IL_16 GM_CSF FIT_3_Ligand SRAGE IL_13 IL_18 G_CSF Fractalkine STNFRI IL_15 IL 1RA IL_10 GIP STNFRII IL_17 IL 2RA IL_12P70 GLP_1 SVEGFR1 IL_1B IL_6 IL_13 Glucagon SVEGFR2 IL_1RA IL_8 IL_15 GRO SVEGFR3 IL_2 IL 9 IL_17 G_CSF TARC IL_21 Insulin IL_1A IFNA2 TNF-α IL_23 IP_10 IL 1B IFNG TPO IL_28A Leptin IL_2 IL_11 TRAIL IL_3 MCP_1MC IL_4 IL_12P40 TSLP IL_4 MIF IL_5 IL_16 VEGF IL_5 MIG IL_7 IL_1A IL_6 MIP_1B LIF IL_29_IFNG1 IL_7 M_CSF MCP_3 IL_33 IL_9 PAI_1 MIP 1A IL_8 I_309 PCT TNF_B INSULIN M_CSF PDGF_BB TPA IP_10 TGFA Rantes LEPTIN TNF-B Resistin LIF XCL1_Lympho SAA MCP_1 SAP MCP_2 SCF MCP_3 SCGF_B MCP_4 SDF_1A MDC TNF-α MIP_1A TRAIL MIP_1B VEGF Visfatin -
aAcceptable performance was defined as (i) being detectable in greater than 25% of the 100 samples on all 3 specimen types and (ii) across-batch CVs of less than 20% for blinded duplicates on at least 2 of the 3 specimen types.
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bMarkers with less than 25% detectability on at least 1 of 3 (serum, heparin plasma, and EDTA plasma) specimen types.
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cMarkers with CVs for across-batch duplicates greater than 20% on 2 or more of 3 specimen types. CVs were calculated for 20 blinded duplicate samples for each specimen type that were placed across different batches.
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- Table 3.
Comparison of assay performance across specimen types and kitsa
Comparison % Detectability number of markersb Median concentration number of markersc Bio-Rad (45 markers) T0 serum vs. T0 heparin plasma Significantly higher in serum 7 16 Significantly higher in plasma 10 19 Similar 21 10 Not evaluabled 7 T3 EDTA plasma vs. T3 heparin plasma Significantly higher in EDTA 2 7 Significantly higher in heparin 2 12 Similar 32 26 Not evaluable 9 Millipore (71 markers) T0 serum vs. T0 heparin plasma Significantly higher in serum 4 18 Significantly higher in plasma 5 13 Similar 51 40 Not evaluable 11 T3 EDTA plasma vs. T3 heparin plasma Significantly higher in EDTA 5 3 Significantly higher in heparin 1 30 Similar 49 38 Not evaluable 16 Bio-Rad vs. Millipore (23 markers) T0 Serum Significantly higher in Bio-Rad 2 15 Significantly higher in Millipore 2 7 Similar 10 1 Not Evaluable 9 T0 heparin plasma Significantly higher in Bio-Rad 4 16 Significantly higher in Millipore 4 7 Similar 6 0 Not evaluable 9 T3 EDTA plasma Significantly higher in Bio-Rad 3 15 Significantly higher in Millipore 3 8 Similar 7 0 Not evaluable 10 -
aComparisons were restricted to markers with acceptable performance—45 Bio-Rad markers, 71 Millipore markers, and 23 markers measured using both kits.
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bComparisons of percentage of detectability across specimen types and kits were conducted using the McNemar's test.
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cComparisons of median observed concentrations across specimen types and kits were conducted using the Wilcoxon signed-rank test. Observations below the assay's lower limit were excluded.
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dMcNemar's P value could not be evaluated because one of the groups had 100% detectability.
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Supplementary Data, Chaturvedi, et al
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Volume 20, Issue 9
