Table 3.

Specificity of Published OspA Primers. OspA Specific Primers Available from the Literature (Reference in Column 1) were Aligned Against 43 OspA Sequences of Different Borrelia Species, Using MEGA 5.0 Software (TAMURA et al 2011). Bold Faced Upper-case Letters (A/T/C/G) Indicate that the Particular Base Matches to All 43 Aligned Sequences While Lower Case Standard Letters (a/t/C/g) Indicate that There is a Mismatch with at Least one Sequence of the Alignment; r/R Means Degenerated Base Position C or T, the Elongation Direction is Indicted by ►or ◄. The Primer Sequences were Taken from the References but Adjusted (i.e. Reversed) where Necessary to Fit for the MEGA Alignment. Primer Names as Given in Column 2 are Taken from the Original Publications. In the Last Column, a Comment is Which Provides Ad-ditional Information about the Specificity, i.e. if A primer Pair was Designed Specifically for a Subset of Borrelia Isolates

Authors [Reference]	Primer Names	Primer Names/Sequences	Comment
Guy & Stanek 1991 [7]	primer pair 1 N1: C1: primer pair 2 N2: C2:	GAgcTtAAAGGAACTTCTGATAA► ◄ACAATTACagTacAAcaaTAc AtGGaTCtGGagtaCTtGAA► ◄AgAaGgaactgtTacTttaag	The primer/probe system is adapted to American B. burgdorferi isolates, to which all oligonucleotides show 100% identity.
Demaerschalck et al. [56]	OspA_fw: OspA_rw:	aATAGGTcTAaTAatAGCCTTAATAGC► ◄ tTtTcAAAGAAGATGgcaaAACAc-TAG	The primers are specific for the three ‘classical’ genospecies, when aligned only to them the number of mismatches is 2 for OspA_fw and 4 for OspA_rw.
Gooskens et al. [17]	BORs: BORas:B OR-TQ (probe):	ATATTTATTGGGaATAGGTcTAaTAT► ◄CTTGTAAGrAAAGAAAAagAcAAaG ◄CTTGTAAGrAAAGAAAAagAcAAaG
Nocton et al. [57]	OspA4: OspA2: OspA3(probe):	CtgcagctTGGaattcaggcacTtc► Ggtcagcagttgaaattacaaaac CagTacAAcaaTAcgACtCaaatGgc	The primer/probe system is adapted to American B. burgdorferi isolates, A3 and A4 show 100% homology, while A2 has two mismatches. The shown primer/probe system is Set 1 in the original publication of 1994
Nocton et al. [57]	OspA149: OspA319: OspA6’(probe) OspA6 (2nd probe):	ATGAAAAAATATTTATTGGGaAT► ◄ ACAGTAGACAAgcTTGAgcTtAAAG GCATGtAAgCAAAATGTTAGC ATTGGGaATAGGTcTAaTATtAGCc	The primer/probe system is also adapted to American B. burgdorferi isolates, A319 has one mismatch, A6 two while A149 and A6’ show 100% homology. Primer/probe system OspA149/319/6’ is Set 2 in the original publication of 1994, while OspA149/319/6 is Set 3
Priem et al. (“ospA”-primers) [28]	outer primer 1 outer primer 2: nested primer 1: nested primer 2:	GGGaATAGGTcTAATATTAGCc► ◄ acTtccACttTAACaaTTagTg gCAAAATGTTAGCagcCTtGAt► ◄ GGAACcAgACTTGAATAcaCAg	The primer/probe system is also adapted to American B. burgdorferi isolates, all but nested 2 show 100% homology, nested 2 displays 2 mismatches.
Priem et al. (“ospA/B”-primers) [28]	outer primer 1: outer primer 2: nested primer 1: nested primer 2:	TTGTAAgCAAAGAAAAaaA► ◄ ttaaaaacGCTTTaAAATAA* GAcGgcAAgTACgatCTAgctG► ◄ ttaaAgAaGgaactgtaacT	The primer/probe system is also adapted to American B. burgdorferi isolates, primer 1 fits 100%, nested 1 shows 3 mismatches, nested 2 one mismatch and primer 2 fits 100% is strain Fort Sheridan 36 is omitted. * Outer primer 2 is shown reversed in the original paper.