Table 1 Primer sequences and qPCR conditions for varATS assay

Oligonucleotide sequences

 Primer-fw (5′–3′)

CCCATACACAACCAATTGGA

 Primer-rev (5′–3′)

TTCGCACATATCTCTATGTCTATCT

 Probe (5′–3′)

6-FAM-TTTTCCATAAATGGT-NFQ-MGB

qPCR reaction conditions (final concentration in qPCR mix)

 Total volume (μL)

20

 DNA volume (μL)

5

 TaqMan^® Gene Expression Mastermix

1×

 Primer (each)

800 nM

 Probe

400 nM

qPCR cycling conditions

 Pre-incubation

2 min–50 °C

 Initial denaturation

10 min–95 °C

 Denaturation

15 s–95 °C

 Annealing and elongation

1 min–55 °C

 Number of cycles

45

Standard material for quantification

gDNA of parasite dilution row

Platform

Real-Time PCR System (Applied Biosystems)