TABLE 2.

Optimization of a quantitative PCR assay for Ehrlichia species (E. canis, E. chaffeensis, E. ewingii, E. muris, and Panola Mountain Ehrlichia spp.) partial sodB gene plasmids^a

Plasmid sodB DNA for:	Melting temp (°C)	Standard curve				LOD (5 copies/reaction)b
		Slope	y intercept	Efficiency (%)	R2	% positive	Average Cq	SD for the Cq variance
E. canis	79.5	−3.544	43.74	92	0.99	100	37.17	0.83
E. chaffeensis	79	−3.578	43.45	90	0.99	100	37.03	0.694
E. ewingii	79	−3.585	42.92	90	0.99	100	35.63	0.969
E. muris	79	−3.479	40.44	94	0.99	85	37.58	1.061
PME	79	−3.486	43.47	94	0.99	95	37.79	1.884

^aStandard curves were determined using the logarithm of plasmid copy numbers/reaction after 10-fold dilutions from 10 to 100,000 copies/reaction.

^bThe limit of detection (LOD) was determined to be 5 copies of plasmid/reaction. Percent positive and the SD of the C_q variance were determined based on positive results from 20 interassay technical replicates.