Table 1.

Method validation.

Validation parameters	Validation results
Minimum Required Dilution	1:10
Clinical cutoff (NR)	0.34
Relative sensitivity using mouse monoclonal ADK6	1.18 µg/mL
Intra-assay precision (ADK6)
QC 1 (3.5 µg/mL)	QC 1: NR 0.010; CV 32.1%
QC 2 (1.5 µg/mL)	QC 2: NR 0.279; CV 13.4%
QC 3 (0.9 µg/mL)	QC 3: NR 0.451; CV 9.3%
Intra-assay precision (pooled PS)
QC 1 (5-fold pooled PS to NCS)	QC 1: NR 0.026; CV 9.2%
QC 2 (10-fold pooled PS to NCS)	QC 2: NR 0.205; CV 6.6%
QC 3 (14-fold pooled PS to NCS)	QC 3: NR 0.389; CV 5.9%
Inter-assay precision (ADK6)
QC 1 (3.5 µg/mL)	QC 1: NR 0.015; CV 7.4%
QC 2 (1.5 µg/mL)	QC 2: NR 0.327; CV 14.3%
QC 3 (0.9 µg/mL)	QC 3: NR 0.409; CV 5.1%
Inter-assay precision (pooled PS)
QC 1 (5-fold pooled PS to NCS)	QC 1: NR 0.027; CV 17.8%
QC 2 (10-fold pooled PS to NCS)	QC 2: NR 0.215; CV 21.2%
QC 3 (14-fold pooled PS to NCS)	QC 3: NR 0.359; CV 12.3%
Selectivity	ADK6 spike into 10 seronegative donors or unspike
	ADK6 at 3.5 µg/mL: 10/10 tested positive
	ADK6 at 0 µg/mL: 10/10 tested negative
Matrix Interference	No interference with hemolysis up to 400 mg/dL
Specificity	ADK8 spiked seronegative serum (NCS)
	NR with ADK8 at 3.5 µg/mL = 0.970
	NR with ADK8 at 0.9 µg/mL = 1.07
Short-term stability	Room temperature: up to tested 4 h
	2–8 °C: up to tested 24 h
	Freeze/thaw: 6 cycles

CV coefficient of variation, NR normalized response, QC quality control, PS positive serum, NCS negative control serum.