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Table 1 Primers and amplification conditions utilized in the current study for the identification and quantification of Legionella spp., Acanthamoeba spp., Naegleria fowleri and Vermamoeba (Hartmannella) vermiformis in pasteurized and unpasteurized tank water samples

From: Molecular detection of Acanthamoeba spp., Naegleria fowleri and Vermamoeba (Hartmannella) vermiformis as vectors for Legionella spp. in untreated and solar pasteurized harvested rainwater

Organism

Primer name

Primer sequence (5'–3')

Gene (size, bp)

Amplification conditions

Reference

Legionella spp. (Identification)

LEG 225

AAGATTAGCCTGCGTCCGAT

16S rRNA (634)

95 °C (1.5 min) followed by 30 cycles of 94 °C (10 s), 64 °C (1 min) and 74 °C (1 min). Final extension: 72 °C (10 min).

Miyamoto et al. [57]

LEG 858

GTCAACTTATCGCGTTTGCT

Legionella spp.

Leg F

CTAATTGGCTGATTGTCTTGAC

23S-5S rRNA (259)

95 °C (1 min) followed by 45 cycles of 95 °C (15 s), 60 °C (15 s) and 72 °C (11 s)

Herpers et al. [58]

Leg R

CAATCGGAGTTCTTCGTG

Acanthamoeba spp.

AcantF900

CCCAGATCGTTTACCGTGAA

18S rDNA (±180)

95 °C (1 min) followed by 45 cycles of 95 °C (15 s), 60 °C (1 min) and 72 °C (40 s)

Qvarnstrom et al. [59]

AcantR1100

TAAATATTAATGCCCCCAACTATCC

Naegleria fowleri

NaeglF192

GTGCTGAAACCTAGCTATTGTAACTCAGT

18S rDNA (153)

95 °C (1 min) followed by 45 cycles of 95 °C (15 s), 64 °C (1 min) and 72 °C (1 min)

Qvarnstrom et al. [59]

NaeglR344

CACTAGAAAAAGCAAACCTGAAAGG

Vermamoeba (Hartmannella) vermiformis

Hv1227F

TTACGAGGTCAGGACACTGT

18S rRNA (502)

95 °C (3 min) followed by 45 cycles of 95 °C (20 s), 58 °C (30 s) and 72 °C (40 s)

Kuiper et al. [60]

Hv1728R

GACCATCCGGAGTTCTCG