Investigation of Borrelia burgdorferi genotypes in Australia obtained from erythema migrans tissue

Figures & data

Figure 1 Erythema migrans patient A at day 3 (biopsy day).

Figure 2 Erythema migrans patient A at day 6.

Table 1 Multiplex primer set

	Wills (University of Newcastle) primers for 16S rRNA gene	Crowder (et al 2010) Osp C gene
Forward	5′ CCC TCA CTA AAC ATA CCT 3′	TGA CGG TAT TTT TA TTT ATA TCT TGT AAT AAT TGA GG 3′
Reverse	5′ ATC TGT TAC CAG CAT GTA AT 3′	5′ TTT GCT TAT TTC TGT AAG ATT AGG CCC TTT 3′

Figure 3 Cladogram of Borrelia species. Source PATRIC as discussed in text. Postion of BLAST findings annotated for Biopsies A–D.

Note: This cladogram was not produced from biopsy material.

Table 2 DNA sequences

Patient A

ACCGTCACTCACTCAAAAGGTAAAGGCTTATTAATATTTTAGATGAGTATGGGGTTGAACATAAGCATTATATTCCAGCTGGAAAACATC TTTTGGTTAGAGATGGAGATGTTGTAAAAGCAGGAGATATGCTTTGTGATGGTAGAATTAATCCTCATGATGTGCTTGAAATTTTAGGTG GGATTATTTTACAAGAATTTCTGCCC

Patient B

TAGTTAATCATTCACAGGTAAAGGCTTATTAATATTTTAGATGAGTATGGGGTTGAACATAAGCATTATATTCCAGCTGGAAAACATCTTT TGGTTAGAGATGGAGATGTTGTAAAAGCAGGAGATATGCTTTGTGATGGTAGAATTAATCCTCATGATGTGCTTGAAATTTTAGGTGGGA TTATTTTACTAGAATTTCTGGACACTTCTTTTTTGCTGTGTTTGGTACCCCTCTCTTTTTTTTTACCCTATAAAACCTATTACGAATCATAAA ATATTATGTTTCTCACGCCATTATTCCCCCCCCCCTCATCGAAACCCTTTTCCCCCCCCTCC

Patient C

CCCTGACAATCAGTTCGAAAGGTAAAGGCTTATTAATATTTTAGATGAGTATGGGGTTGAACATAAGCATTATATTCCAGCAGGAAAACA TCTTTTGGTTAGAGATGGAGATGTTGTAAAAGCAGGAGATATGCTTTGTGATGGTAGAATTAATCCTCATGATGT

Patient D

ACAGCGTCAGGACCTCGGCTGAGTGATAGGTGTATTTAGATCTTTCCCTGAATTATTACAGATATAGATAAAAATACCGTCAA

Figure 4 Distance tree of results A and B using neighbor joining construction.

Figure 5 Distance tree of results C using neighbor joining construction.

Table 3 Top scoring sequences

Accession	Description	Max score	Total score	Query coverage	E value	Max ident
Patient A
CP002228.1	Borrelia burgdorferi N40, complete genome	339	339	91%	6.00E-90	99%
CP001205.1	Borrelia burgdorferi ZS7, complete genome	339	339	91%	6.00E-90	99%
AE000783.1	Borrelia burgdorferi B31, complete genome	339	339	91%	6.00E-90	99%
CP002312.1	Borrelia burgdorferi JD1, complete genome	333	333	91%	3.00E-88	98%
Patient B
CP002228.1	Borrelia burgdorferi N40, complete genome	327	327	55%	2.00E-86	98%
CP001205.1	Borrelia burgdorferi ZS7, complete genome	327	327	55%	2.00E-86	98%
AE000783.1	Borrelia burgdorferi B31, complete genome	327	327	55%	2.00E-86	98%
CP002312.1	Borrelia burgdorferi JD1, complete genome	322	322	55%	1.00E-84	98%
Patient C
CP002228.1	Borrelia burgdorferi N40, complete genome	267	267	95%	2.00E-68	97%
CP001205.1	Borrelia burgdorferi ZS7, complete genome	267	267	95%	2.00E-68	97%
AE000783.1	Borrelia burgdorferi B31, complete genome	267	267	95%	2.00E-68	97%
CP002312.1	Borrelia burgdorferi JD1, complete genome	261	261	95%	1.00E-66	96%

Table 4 Paired alignment patients A B

Table 5 Paired alignment patients A C

Table 6 Demographic data and clinical details

	Patient
	A	B	C	D
Gender	Male	Female	Female	Male
Overseas travel	Yes	Never	Yes	Never
Geographic	152.8E	152.7E	151.3E	152.8E
coordinates	31.66S	31.73S	33.74S	31.32S
Systemic illness	No	Yes	Very severe flu-like illness developed on 5th day, including fever and meningism with severe headache worse with coughing and shaking of the head, photophobia and retro-orbital pain. She developed no detectable neurological deficit	No
Treatment	Doxycyline	Doxycyline	Amoxycillin plus clavulanate	Doxycyline
Dose	400 mg daily	400 mg daily	875/125 mg bd	400 mg daily
Cautions			Breast feeding
Recovery factors	Efficient progress	Supply interruption at 1 week for 1 week and developed flu-like symptoms. Therapy resumed and progress good	Failed response. Eventually stopped breast feeding at 8 weeks, and doxycycline 400 mg daily added. Within days, changed to clarithromycin because of nausea. Response ensued	Efficient progress
Recovery time	4 weeks	8 weeks	12 weeks	4 weeks
Follow-up testing at 19 weeks Igenex, Palo Alto, USA
Borrelia burgdorferi IgA, G, and M			Negative
B. burgdorferi multiplex PCR			Negative
Babesia microtii and Babesia duncanii			Negative
Bartonella henselae			Negative