Fluorescence in situ Hybridization method using Peptide Nucleic Acid probes for rapid detection of Lactobacillus and Gardnerella spp.

António Machado; Carina Almeida; Débora Salgueiro; Ana Henriques; Mario Vaneechoutte; Freddy Haesebrouck; Maria João Vieira; Ligia Rodrigues; Nuno Filipe Azevedo; Nuno Cerca

doi:10.1186/1471-2180-13-82

BMC Microbiology

Table 1 Bacterial strains used in PNA-FISH assays and their specificity with Lac663 and Gard162 probes

From: Fluorescence in situ Hybridization method using Peptide Nucleic Acid probes for rapid detection of Lactobacillus and Gardnerella spp.

Bacterial species	Collection strain	Lac663 Probe efficiency	Gard162 Probe efficiency
Lactobacillus acidophilus	ATCC 4356^T	++++	-
L. crispatus	ATCC 33820^T	++++	-
L. gasseri	ATCC 9857^T	++++	-
L. reuteri	NCFB 2656^T	+++	-
L. rhamnosus	ATCC 7469^T	++++	-
L. rhamnosus	CECT 288^T	++++	-
L. johnsonii	ATCC 11506^T	++++	-
L. hilgardii	NCFB 962^T	+++	-
L. delbrueckii subsp. delbrueckii	ATCC 9649^T	+++	-
L. delbrueckii subsp. lactis	ATCC 12315^T	+++	-
L. pentosus	CECT 4023^T	++++	-
L. casei	CECT 5275^T	++++	-
L. coryniformis subsp. torquens	CECT 4129^T	++++	-
L. paracasei	CECT 227^T	++++	-
L. agilis	CCUG 31450^T	++++	-
L. animalis	ATCC 35046^T	+++	-
L. bifermentans	ATCC 35409^T	+++	-
L. brevis	ATCC 14869^T	++++	-
L. buchneri	ATCC 4005^T	+++	-
L. fermentum	ATCC 11739^T	+++	-
L. curvatus subsp. curvatus	ATCC 25601^T	++++	-
L. farciminis	DSM 20182T	++++	-
L. fructivorans	ATCC 8288^T	+++	-
L. gallinarum	CCUG 31412^T	++++	-
L. graminis	DSM 20719^T	++	-
L. hamsteri	ATCC 43851^T	+++	-
L. helveticus	ATCC 15009^T	++++	-
L. intestinalis	ATCC 49335^T	+++	-
L. murinus	ATCC 35020^T	++++	-
L. parabuchneri	ATCC 12936^T	++++	-
L. paracasei subsp. paracasei	CCUG 27320^T	+++	-
L. plantarum	NCIMB 8827^T	+++	-
L. ruminis	ATCC 27781^T	++++	-
L. sakei subsp. carnosus	CCUG 8045^T	++	-
L. salivarius	DEVRIESE 94/438^T	+++	-
L. plantarum	NCCB 46043^T	+++	-
L. lactis 53	-	-	-
Streptococcus. thermophilus A	-	-	-
S. thermophilus B	-	+++	-
Leuconostoc mesenteroides	-	-	-
Bacillus subtilis	DSM 7-10^T	-	-
Enterococcus faecium	CECT 410^T	-	-
E. faecalis	CECT 184^T	-	-
Gardnerella vaginalis 5-1	-	-	++++
G. vaginalis 101	-	-	++++
G. vaginalis AMD	-	-	++++
G. vaginalis	ATCC	-	++++
G. vaginalis	Belgian isolate 1	-	+++
G. vaginalis	Belgian isolate 2	-	++++
G. vaginalis	Belgian isolate 3	-	++++
G. vaginalis	Belgian isolate 4	-	++++
G. vaginalis	Belgian isolate 5	-	++++
G. vaginalis	Belgian isolate 6	-	++++
G. vaginalis	Belgian isolate 7	-	+++
G. vaginalis	Belgian isolate 8	-	+++
G. vaginalis	Belgian isolate 9	-	++++
G. vaginalis	Belgian isolate 10	-	++
G. vaginalis	Belgian isolate 11	-	++++
G. vaginalis	Belgian isolate 12	-	+++
G. vaginalis	Belgian isolate 13	-	+++
G. vaginalis	Belgian isolate 14	-	++
G. vaginalis	Belgian isolate 15	-	+++
G. vaginalis	Belgian isolate 16	-	+++
G. vaginalis	Belgian isolate 17	-	++++
G. vaginalis	Belgian isolate 18	-	++++
Atopobium vaginae	CCUG 38953^T	-	-
A. vaginae	CCUG 42099^T	-	-
A. vaginae	CCUG 44116^T	-	-
A. vaginae	Clinical isolate	-	-
Bacillus cereus	-	-	-
Enterobacter aerogenes	CECT 684^T	-	-
Escherichia coli O157:H7	NCTC 12900^T	-	-
Staphylococcus aureus	CECT 976^T	-	-
S. aureus	CECT 86^T	-	-
Shigella flexneri	ATCC 12022^T	-	-
Listeria monocytogenes	-	-	-
L. monocytogenes	CECT 5873^T	-	-
L. seeligeri	CECT 917^T	-	-
Klebsiella pneumoniae subsp. ozaenae	ATCC 11296^T	-	-
Salmonella Typhi	-	-	-
S. enterica	-	-	-
Escherichia coli	CECT 434^T	-	-
Prevotella bivia	ATCC 29303^T	-	-
Mobiluncus mulieris	ATCC 26-9^T	-	-
Fusobacteria nucleatum	Clinical isolate	-	-

The PNA Probe (Lac663 and Gar162) efficiencies were tested in triplicate experiments for each strain, with the following hybridization PNA FISH qualitative evaluation: (−) Absence of hybridization; (++) Moderate hybridization; (+++) Good hybridization; (++++) Optimal hybridization. The table shows the median value from the three experiments for each strain.

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