TABLE 3

Primers and probes used in reverse line blots

Target species for primer or probeaTarget geneNucleotide sequence (5′→3′)bReferencec
Primers
    Borrelia burgdorferi sensu lato5S-23S IGSACC ATA GAC TCT TAT TAC TTT GAC CA24
Biotin-GAG AGT AGG TTA TTG CCA GGG24
TCA GGG TAC TTA GAT GGT TCA CTT25
Biotin-GAG TTC GCG GGA GAG TAG GTT ATT25
    Rickettsia spp.16S rRNAGAA CGC TAT CGG TAT GCT TAA CAC A27, modified from reference 26
Biotin-CAT CAC TCA CTC GGT ATT GCT GGA27, modified from reference 26
23S-5S IGSGAT AGG TCR GRT GTG GAA GCA C28
Biotin-TCG GGA YGG GAT CGT GTG TTT C28
    Anaplasma/Ehrlichia spp.16S rRNAGGA ATT CAG AGT TGG ATC MTG GYT CAG29
Biotin-CGG GAT CCC GAG TTT GCC GGG ACT TYT TCT84
    Babesia/Theileria spp.18S rRNAGAC ACA GGG AGG TAG TGA CAA G85
Biotin-CTA AGA ATT TCA CCT CTG ACA GT85
    Coxiella burnetiihtpABTAT GTA TCC ACC GTA GCC AGT C78
Biotin-CCC AAC AAC ACC TCC TTA TTC78
Probes, membrane 1
    B. burgdorferi sensu lato5S-23S IGSCTT TGA CCA TAT TTT TAT CTT CCA24
    B. burgdorferi sensu lato 2C TTC CAT CTC TAY TTT GCC AATThis paper
    B. burgdorferi sensu strictoAAC ACC AAT ATT TAA AAA ACA TAA24
    B. afzeliiAAC ATT TAA AAA ATA AAT TCA AGG24
    B. garinii/B. bavariensisAAC ATR AAC ATC TAA AAA CAT AAA24, modified
    B. spielmaniiGTC AAT ATC TAT TTT CTT TTT TAT GThis paper
    B. valaisiana 2CAT GTC AAT ATC TAT TTT ATT TTT TAC ATT AThis paper
    B. valaisiana (VS116)CAT TAA AAA AAT ATA AAA AAT AAA TTT AAG G24
    B. valaisiana (VSNE)TAT ATC TTT TGT TCA ATC CAT GT86
    B. lusitaniaeTTT TTA AAT CAA ACA TTC AAA AAA ATThis paper
    B. lusitaniae (LusiNE)TCA AGA TTT GAA GTA TAA AAT AAA A86
    B. lusitaniae (LusiNE1)CAT TCA AAA AAA TAA ACA TTT AAA AAC AT31
    B. lusitaniae (LusiNE2)AAA TCA AAC ATT CAA AAA AAT AAA C31
    B. bissetti/B. carolinensisCAC TAA CAT TTA AAA AAT ATA AAA TAA AATThis paper
    Rickettsia spp.23S-5S IGSTAG CTC GAT TGR TTT ACT TTG28
    SFG RickettsiaeACT CAC AAR GTT ATC AGG T28
    Typhus group rickettsiaeGTT ATT CTA TCG TTT TAT GTY ACG28
    R. belliiGTG TTT ATT CTA TAA TAT GTC AG28
    R. conoriiGTT ATA TAC TGT AGC CCT G28
    R. aeschlimanniiATA TTA TAC TGT ATG TAG CCC C28
    R. rickettsii-R. sibiricaGTT ATA CTG TAG TCC TGC AA28
    R. slovacaGTA GCC CCT GCC ACG ATA28
    R. helveticaCAT GGC TTG ATC CAC GGT A28
    R. raoultiiTCA ACT AAT AAA TTT GCT GAG TA81
    R. monacensisCAA TGT CAT ACC GTG GTC AAGM. Wijnveld, unpublished
    C. burnetiihtpABGCA AGA ATA CGG ACT CAC GA77
Probes, membrane 2
    Ehrlichia/Anaplasma spp.16S rRNAGGG GGA AAG ATT TAT CGC TA84
    A. phagocytophilum 1TTG CTA TAA AGA ATA ATT AGT GG29
    A. phagocytophilum 3TTG CTA TGA AGA ATA ATT AGT GG29
    A. phagocytophilum 5TTG CTA TAA AGA ATA GTT AGT GGProtocol book UCTD
    A. phagocytophilum 7TTG CTA TAG AGA ATA GTT AGT GGProtocol book UCTD
    “Ca. Neoehrlichia mikurensis”GCT GTA GTT TAC TAT GGG TA29
    Theileria/Babesia catch-all18S rRNATAA TGG TTA ATA GGA RCR GTT G87
    Babesia catch-all 1ATT AGA GTG TTT CAA GCA GAC87
    Babesia catch-all 2ACT AGA GTG TTT CAA ACA GGC87
    B. divergensACT RAT GTC GAG ATT GCA C76
    Theileria (Babesia) microtiGRC TTG GCA TCW TCT GGA87
    B. canis canisTGC GTT GAC GGT TTG AC88
    B. canis canis 2TGG TTG GTT ATT TCG TTT TCG87
    Babesia venatorumCGA TTT CGC TTT TGG GAT T27
    Theileria catch-allATT AGA GTG CTC AAA GCA GGC87
    Rickettsia spp.16S rRNATTT AGA AAT AAA AGC TAA TAC CG26
    R. conoriiCTT GCT CCA GTT AGT TAG T26
    R. helveticaGCT AAT ACC ATA TAT TCT CTA TG26
    R. massiliaeTGG GGC TTG CTC TAA TTA GT89
    R. raoultiiCTA ATA CCG CAT ATT CTC TAC G27
  • a All probes were labeled with a C6 amino linker for covalent binding to the membranes.

  • b Bold letter indicates the difference from the original probe: nucleotide ambiguity code uses R to represent A or G to cover more species.

  • c Protocol book UCTD, oligonucleotide sequences obtained from the Utrecht Center for Ticks and Tick-borne Diseases Summer School Protocol Book (2012), no original reference available.