Quantitative PCR and Digital PCR for Detection of Ascaris lumbricoides Eggs in Reclaimed Water

Table 1

Detection of A. lumbricoides by qPCR from reclaimed water seeded with different amounts of A. lumbriocoides eggs.

Protocol  1 (for 500 mL)			Protocol  2 (for 10 L)
Number of eggs	Mean; copies μL^−1	Result	Number of eggs	Mean; ITS-1 copies μL^−1	Positive extractions	Result
1		Presence	10	—	—/6	Absence
1		Presence	10		2/6	Presence
1		Presence	10	—	—/6	Absence
1		Presence	10		2/6	Presence
1		Presence	10		1/6	Presence
Average ± SD
5		Presence	20		4/6	Presence
5		Presence	20		2/6	Presence
5		Presence	20		2/6	Presence
5		Presence	20		1/6	Presence
5		Presence	20		3/6	Presence
Average ± SD
10		Presence	50		6/6	Presence
10		Presence	50		2/6	Presence
10		Presence	50		1/6	Presence
10		Presence	50		3/6	Presence
10		Presence	50		5/6	Presence
Average ± SD

The averages are for three aliquots in 500 mL and for three aliquots of each of six different extractions per sample in 10 L. Positive extractions: number of positive extractions in each batch. Only from the “positive” aliquots were included in the calculation. Presence: Ct < 37. Absence: Ct > 37. Protocol  2, below 10-egg amplification was not observed.