Table 2.

Evaluation of the new dual priming oligonucleotides

Sample no.d	Specimen	PCRe	Findings (based on sequence)f	Fg	Rg	Culture
1	Abscess, lung	Neg	Bacterial backgroundb,c	✓	✓	Aspergillus sp.
2	Tissue, prosthetic joint	Neg	Bacterial backgrounda	✓	✓	No growth
3	Abscess, bone	Pos	Kingella kingaea,b,c	✓	✓	Kingella kingae
4	Abscess, brain	Pos	Streptococcus intermediusa,b	✓	✓	Streptococcus intermedius
5	Abscess, kidney	Pos	Escherichia colia,b,c	✓	✓	Escherichia coli
6	Abscess, liver	Pos	Enterococcus faecium, Escherichia coli	✓	✓	Enterococcus faecium, Escherichia coli
7	Abscess, subcutaneous	Pos	Enterococcus faecium, Finegoldia magnaa	✓	✓	Enterococcus faecium, Finegoldia magna
8	Bile	Pos	Streptococcus anginosus	✓	✓	Streptococcus anginosus
9	Pleural fluid	Pos	Streptococcus pyogenesa,b	✓	✓	Streptococcus pyogenes
10	Tissue	Pos	Staphylococcus epidermidis	✓	✓	Staphylococcus epidermidis
	Pacemaker pouch		Propionibacterium acnesa,b	✓	✓
11	Tissue prosthetic joint	Pos	Staphylococcus aureusb	✓	✓	Staphylococcus aureus
12	Tissue prosthetic joint	Pos	Bacteroides fragilisa,b	✓	✓	Bacteroides fragilis
13	Tissue subcutaneous	Pos	Morganella morganii	✓	✓	Morganella morganii
14	Abscess, aorta	Neg	Bacterial background	✓	✓	No growth
15	Abscess, neck	Neg	Bacterial background	✓	✓	No growth
16	Abscess, presacral	Neg	Propionibacterium acnes (high), bacterial background (low)	✓	✓	No growth
17	Abscess, subcutaneous	Neg	Bacterial background	✓	✓	No growth
18	Heart valve	Neg	Staphylococcus aureus (high), bacterial background (low)	✓	✓	Staphylococcus aureus
19	Heart valve	Neg	Bacterial background	✓	✓	No growth
20	Peritoneal fluid	Neg	Escherichia coli	✓	✓	Escherichia coli
21	Pleural fluid	Neg	Streptococcus intermedius (high), bacterial background (low)	✓	✓	No growth
22	Pleural fluid	Neg	Bacterial background	✓	✓	No growth
23	Subdural hematoma	Neg	Bacterial background	✓	✓	No growth
24	Synovial fluid	Neg	Bacterial background	✓	✓	No growth
25	Tissue, prosthetic joint	Neg	Staphylococcus aureus (high), bacterial background (low)	✓	✓	Staphylococcus aureus
26	Tissue, surgical wound	Neg	Staphylococcus aureus (high), bacterial background (low)	✓	✓	Staphylococcus aureus
27	Abscess, abdominal	Pos	Haemophilus parahaemolyticus	✓	✓	No growth
28	Abscess, brain	Pos	Streptococcus intermedius, Fusobacterium nucleatum	✓	✓	Streptococcus intermedius
29	Abscess, brain	Pos	Fusobacterium nucleatum	✓	✓	No growth
30	Abscess, brain	Pos	Aggregatibacter aphrophilus	✓	✓	Aggregatibacter aphrophilus
31	Abscess, hip	Pos	Bacteroides fragilis	✓	✓	Bacteroides fragilis
32	Abscess, kidney	Pos	Citrobacter koseri	✓	✓	Citrobacter koseri
33	Abscess, liver	Pos	Propionibacterium acnes	✓	✓	Propionibacterium acnes
34	Abscess, liver	Pos	Escherichia coli, Clostridium perfringens	✓	✓	Escherichia coli
35	Abscess lung	Pos	Lactobacillus gasseri	✓	✓	Lactobacillus gasseri
36	Abscess pancreas	Pos	Neisseria sp.	✓	✓	No growth
37	Abscess pancreas	Pos	Mycobacterium tuberculosis	✓	✓	Mycobacterium tuberculosis
38	Abscess retroperitoneal	Pos	Enterococcus faecium, Klebsiella pneumoniae	✓	✓	Enterococcus faecium
39	Abscess shoulder	Pos	Finegoldia magna	✓	✓	Finegoldia magna
40	Abscess subcutaneous	Pos	Streptococcus dysgalactiae	✓	✓	Streptococcus dysgalactiae
41	Bile	Pos	Escherichia coli	✓	✓	Escherichia coli
42	Biopsy bone	Pos	S. aureus	✓	✓	Staphylococcus aureus
43	Biopsy bone	Pos	Enterobacter asburiae/hormaecei	✓	✓	Enterobacter sp.
44	Biopsy bone	Pos	Staphylococcus aureus, Staphylococcus epidermidis	✓	✓	Staphylococcus aureus
45	Heart blood	Pos	Clostridium septicum	✓	✓	Clostridium septicum
46	Pleural fluid	Pos	Escherichia coli	✓	✓	Escherichia coli
47	Pleural fluid	Pos	Staphylococcus intermedius	✓	✓	Staphylococcus intermedius
48	Pleural fluid	Pos	Prevotella pleuritidis	✓	✓	Eikenella corrodens, Campylobacter gracilis
49	Tissue, mediastinum	Pos	Enterococcus faecalis	✓	✓	Enterococcus faecalis
50	Tissue, prosthetic joint	Pos	Staphylococcus epidermidis	✓	✓	Staphylococcus epidermidis

^aCross-reactivity with human DNA observed for Bosshard primers.

^bCross-reactivity with human DNA observed for CLSI primers.

^cCross-reactivity with human DNA observed for the Bosshard forward/CLSI reverse primer combination.

^dSamples consist of 15 PCR-negative samples (1 to 2 and 14 to 26) and 35 PCR-positive samples (3 to 13 and 27 to 50). Samples 1 to 13 were used for the initial primer evaluation that also included the conventional primer pairs.

^ePos, positive; Neg, negative.

^fBacterial background indicates for the most part mixed chromatograms dominated by Pseudomonas spp.

^gF, forward sequence; R, reverse sequence. The check mark indicates that no unexpected fragments/cross-reactivity was observed.