Validation of a pan-orthopox real-time PCR assay for the detection and quantification of viral genomes from nonhuman primate blood

Table 5 Summary of findings for Orthopox DNA PCR

Parameter	Conclusion
Precision	Acceptable when GC/5 μL is ≥ LLOQ
Repeatability
Intermediate
Selectivity	Matrix effects of 13–25%
Accuracy	within 50–150% (when ≥ LLOQ)
Standard Curve	LOD: 2.5 GC/5 μL
	LLOQ: 50 GC/5 μL
	ULOQ: 5 × 10⁷ GC/5 μL
	Efficiency: 2.0 ± 0.2
	Stability: ≥ 92 days
Stability
HA standards	stable after 10 freeze thaws
PEC	stable for at least 4 months
Master Mix	stable for at least 8 months
Specificity	orthopox specific; see also [4]
Robustness	no/little effect when changing extraction kit lots, Master Mix lots, or LightCycler instruments

ISSN: 1743-422X