Table 1.

Sonication time trials analyzed for the optimal detection of Pseudoloma neurophilia. Replicate spiked water filters at 58,750 spores/L were sonicated at 1, 2, 5, 8, and 10 min and validated using qPCR, with values reported as Ct values. Extracts from the sonication time trial were used to optimize dPCR parameters, in which replicate values are reported in copy numbers/μL. The Qiagen standard priming profile was employed (2 min at 95 ° C and 40 cycles of 95 °C for 15 s, then 60 °C for 30 s).

	Optimal Sonication Time (qPCR)				dPCR Validation
	Filter 1		Filter 2		Filter 1		Filter 2
Time	Replicate 1	Replicate 2	Replicate 1	Replicate 2	Replicate 1	Replicate 2	Replicate 1	Replicate 2
1 min	31.84	31.46	31.9	32.31	6.4	5	4.4	3.2
2 min	30.54	30.95	30.52	30.84	7.7	9.9	11.7	7.9
5 min	32.29	32.27	33.44	34.4	4.1	2.9	1	1.2
8 min	30.63	30.22	29.57	29.51	10	8.4	32.1	33.7
10 min	31.15	31.6	29.54	29.62	7.7	8.1	20.8	14.5