Effective Concentration and Detection of Cryptosporidium, Giardia, and the Microsporidia from Environmental Matrices

Table 1

Comparison of Method 1623 with CFD-PCR results of spiking trials using 100 C. parvum oocysts and 100 G. intestinalis cysts and about 1000 spores of E. bieneusi and E. intestinalis, respectively.

	Batch number	Method 1623; Sample data				C. parvum + G. intestinalis			CTD-qPCR; Sample data				C. parvum + G. intestinalis			E. intestinalis			E. bieneusi
		Replicate number	Volume (L)	Turbidity (NTUs)	Number of filters used	Number of (oo)cysts spiked^a	Number of C.  parvum detected^b	Number of G. intestinalis detected^b	Replicate number	Volume (L)	Turbidity (NTUs)	Number of filters used	Number of (oo)cysts spiked^a	Number of C. parvum detected^c	Number of G. intestinalis detected^c	Number of E.  intestinalis spiked^d	Number of E. intestinalis detected^c	Percent recovery^e	Number of E. bieneusi spiked^d	Number of E. bieneusi detected^c	Percent recovery^e
Trial 1	1	1	26.5	1.1	1	100	25	25	1	20	1.1	1	100	31	14		674	64		523	42
		2	22.5	1.3	1	100	10	6	2	20	1.1	1	100	17	35		583	56		788	64
		3	24.8	1.1	1	100	16	21	3	20	1.1	1	100	63	21		616	59		532	43
	2	1	21.0	7.1	1	100	4	10	1	20	7.1	1	100	17	8		512	49		505	41
		2	20.8	7.1	1	100	2	30	2	20	7.1	1	100	63	5		338	32		343	28
		3	20.3	7.1	1	100	0	22	3	20	7.1	1	100	12	18		311	30		689	56
	3	1	30.0	54.0	2	100	17	18	1	20	54.0	2	100	49	59		441	42		423	34
		2	23.0	54.0	2	100	3	1	2	20	54.0	2	100	37	15		534	51		510	41
		3	22.8	54.0	1	100	5	3	3	20	54.0	2	100	9	0		606	58		413	34
		MB	20.7	54.0	1	0	0	2	MB	20	54.0	2	0	0	0		0	0		0	0
Trial 2	1	1	21.8	0.2	1	100	25	27	1	22	0.2	1	100	34	28		788	70		576	62
		2	20.3	0.2	1	100	40	20	2	22	0.2	1	100	35	26		702	62		412	44
		3	22.2	0.2	1	100	28	14	3	22	0.2	1	100	34	0		744	66		124	13
	2	1	23.0	0.3	1	100	73	57	1	22	0.3	1	100	28	22		712	63		431	46
		2	21.3	0.3	1	100	44	58	2	22	0.3	1	100	5	7		211	19		346	37
		3	20.3	0.3	1	100	45	37	3	22	0.3	1	100	16	24		245	22		378	40
	3	1	21.0	3.8	1	100	49	52	1	22	3.8	1	100	6	11		ND	NA		ND	NA
		2	23.0	3.8	1	100	48	46	2	22	3.8	1	100	27	17		399	35		145	16
		3	21.8	3.8	1	100	50	41	3	22	3.8	1	100	16	23		204	18		185	20

Number of C. parvum and G. intestinalis (oo)cysts spiked using AccuSpike-IR suspensions.
^bNumber of (oo)cysts detected in entire sample.
^cEstimated number of pathogens in sample (number of pathogens calculated × total volume of gDNA)/(volume of gDNA analyzed).
^dEstimated number of spores spiked based on hemocytometer counts.
^eEstimated percent recovery (number of spiked/number of detected × 100).
ND = not done; NA = not applicable; MB = method blank.