Table 7.

Free-living amoebae primers and probes. 23S pan-Legionella is conserved by all species of Legionella, but the probes were uniquely designed to specifically identify each of these species: L. micdadei, L. anisa, L. bozemanii, and L. longbeachae. All primers and probes were designed and validated previously by the authors listed in the references below.

Target Species	Primer/Probe Name	Primer/Probe Sequence	Amplicon Length (bp)	Reference
Acanthamoeba spp.	18S rRNAF 18S rRNAR 18S rRNAP	5′-CGACCAGCGATTAGGAGACG-3′ 5′-CCGACGCCAAGGACGAC-3′ 5′-FAM-TGAATACAAAACACCACCATCGGCGC-BHQ1-3′	63	[65]
N. fowleri	ITSF ITSR ITSP	5′-GTGAAAACCTTTTTTCCATTTACA-3′ 5′-AAATAAAAGATTGACCATTTGAAA-3′ 5′-HEX-GTGGCCCACGACAGCTTT-BHQ1-3′	69	[66,67]
Legionella species	23SF 23SR 23SP	5′-CCCATGAAGCCCGTTGAA-3′ 5′-ACAATCAGCCAATTAGTACGAG TTAGC-3′ 5′-HEX-TCCACACCTCGCCTATCAACGTCGTAGT-BHQ1-3′	92	[68]
L. pneumophila	mipF mipR mipP	5′-AAAGGCATGCAAGACGCTATG-3′ 5′-GAAACTTGTTAAGAACGTCTTTCATTTG-3′ 5′-FAM-TGGCGCTCAATTGGCTTTAACCGA-BHQ1-3′	78	[68]
L. micdadei L. anisa L. bozemanii L. longbeachae	Pan-Legionella F Pan-Legionella R LmicdadeiP LanisaP Lbozemanii LlongbeachaeP	5′-GTACTAATTGGCTGATTGTCTTG-3′ 5′-TTCACTTCTGAGTTCGAGATGG-3′ 5′-FAM-AGCTGATTGGTTAATAGCCCAATCGG-BHQ1-3′ 5′-HEX-CTCAACCTACGCAGAACTACTTGAGG-BHQ1-3′ 5′-FAM-TACGCCCATTCATCATGCAAACCAGnT-BHQ1-3′ 5′-HEX-CTGAGTATCATGCCAATAATGCGCGC-BHQ1-3′	Not available	[69]