Table 3

Primers and amplification conditions used in the study

Forward primer (5'-3')
Reverse primer (5'-3')
Specific for
Annealing temperature/time
MgCl2 concentration (mmol/l)
Size of product (bp)

AGTAGTTTACTACTTTGCCG
ACTGCTGCCTCCCGTAGGAG
Universal (R1 and R2)
58°C/60 secs
1.5
350
CATAACGTCGCAAGACCAAA
GTGCAATATTCCCCACTGCT
E. coli
58°C/45 secs
1.5
187
TTGGGAATAACGGTTGGAAA
TGTCTCAGTCCCAGTGTTGG
Chlamydia spp.
59°C/30 secs
1.5
203
CGCACGGGTGAGTAAGGTA
GCGTCATAGCCTTGGTAAGC
Campylobacter spp.
66°C/1 min
2.5
170
AGTAGTTTACTACTTTGCCG
CCGATGGCGTGAGGCCCTAA
Yersinia spp.
65°C/30 secs
3.5
154
CGCACGGGTGAGTAAGGTA
GCTTAACACAAGTTGACTAG
Campylobacterjejuni
63°C/30 secs
2.5
70
CGCACGGGTGAGTAAGGTA
GTCTTACATAAGTTAGATA
Campylobacter concisus
55°C/30 secs
2.0
70
CGCACGGGTGAGTAAGGTA
ATACCTCATACTCCTATTTAAC
Bacteroides ureolyticus
55°C/30 secs
2.0
70

Specific oligonucelotides were designed as described here. Standard reaction mix was used with each set of oligonucleotides (see Materials and method), although annealing conditions varied, as did MgCl2 concentration. Denaturation conditions were always 94°C for 1 min and extension was performed at 72°C for 1 min (10 min final extension). Typically 35–40 cycles were performed. If no PCR product was detected, then a second round of amplification was performed using 2 μl of PCR product as template.

Cox et al. Arthritis Res Ther 2003 5:R1   doi:10.1186/ar602

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