Table 1.
Primers and probes for multiplex real-time and HRM detection/typing of nonpneumophila Legionella spp.
| Primer/probe | Sequence (5′→3′) | Species target | Gene target | GenBank accession no. |
Primer/probe final concentration |
|---|---|---|---|---|---|
| bozemanii-F bozemanii-R bozemanii-P3 |
TCCGCTGCTGAAGTGATTATG CATGCAAACCACCCGATACT Q705-AAATTTACCACCGGCGTGAAGCAC-BHQ3 |
L. bozemanii, L. anisa, L. parisiensis, L. tucsonensis sg 1 and 2 |
gyrB | HQ717438 | 125 nmol/L 125 nmol/L 25 nmol/L |
| dumoffii-F dumoffii-R dumoffii-P |
CAGGAAAGCGCGACATCTAT ATCCAGCTCGTTCGCAATAA HEX-TGGAAACCCTCAATGGTCCGTTCT-BHQ1 |
L. dumoffii | legS2 | EU107519 | 250 nmol/L 250 nmol/L 50 nmol/L |
| feeleii-F feeleii-R feeleii-P2 |
AACCGGTTTATCGGTCTTT ATCAACCAGCTTGTCTCG TEX615-AGCTTGAGAATTTGATGGATTATCACTCGC-BHQ2 |
L. feeleii | figA | AY753535 | 125 nmol/L 125 nmol/L 25 nmol/L |
| LLB-F1 LLB-R1 LLB-P1 |
CTGCAGAAGTTGCTGATTGTG GACGTGGCGAATGACTTATCT Q670-TGTCGCCAAGAAGTTGTATCTCATGCT-BHQ3 |
L. longbeachae, L. sainthelensis sg 1 and 2 |
ligB | AY512558 | 125 nmol/L 125 nmol/L 25 nmol/L |
| micdadei-F micdadei-R micdadei-P |
TGACAAGTGAGAGCAAGAGTT GTATCTATTCCGACAGCGATAGG FAM-ACAGAAGGAGAACCTTCCGGTGTG-BHQ1 |
L. micdadei | migB | AY512559 | 125 nmol/L 125 nmol/L 25 nmol/L |
| LLB-HRM-F LLB-HRM-R |
TGGCTAAGCGTAGTGCTG CAACAGTTACAGTATCTGATTTAC |
L. longbeachae | mip | X83036 | 250 nmol/L 250 nmol/L |
| bozemanii-HRM-F1 bozemanii-HRM-R1 |
CCAAGCGGCTTGCAATATAA CAAATACAGTACCATCAATTAAAGTACC |
L. bozemanii | mip | U91609 | 250 nmol/L 250 nmol/L |
| feeleii-HRM-F1 feeleii-HRM-R |
GACCTAATGGCAAAACGAAATG CGCCTTCTTTGGCCTTATTC |
L. feeleii | mip | U92205 | 250 nmol/L 250 nmol/L |
| tmRNA-HRM-F tmRNA-HRM-R |
GGCGACCTGGCTTC GGTCATCGTTTGCATTTATATTTA |
L. anisa, L. parisiensis, L. tucsonensis sg 1 and 2 |
ssrA | HG525464 | 250 nmol/L 250 nmol/L |