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

Table 3 Results of the Lac663 and Gard162 probes specificity test in artificial mixed samples

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

Species in the artificial mixed samples	Bacteria strain collection codes	Multiplex PNA-FISH assay
Species in the artificial mixed samples	Bacteria strain collection codes	Lac663 Probe efficiency	Gard162 Probe efficiency
L. pentosus;	CECT 4023^T; -	++++	++++
G. vaginalis 51	CECT 4023^T; -	++++	++++
L. casei;	CECT 5275^T; -	++++	++++
G. vaginalis 101	CECT 5275^T; -	++++	++++
L. rhamnosus;	CECT 288^T; -	++++	++++
G. vaginalis AMD	CECT 288^T; -	++++	++++
L. crispatus;	ATCC 33820^T; -	++++	++++
G. vaginalis ATCC	ATCC 33820^T; -	++++	++++
L. delbrueckii sub. delbrueckii; Atopobium vaginae	ATCC 9649^T; CCUG 38953^T	+++	-
L. acidophilus;	ATCC 4356^T; CCUG 42099^T	++++	-
A. vaginae	ATCC 4356^T; CCUG 42099^T	++++	-
L. gasseri;	ATCC 9857^T; CCUG 44116^T	++++	-
A. vaginae	ATCC 9857^T; CCUG 44116^T	++++	-
L. paracasei sub. paracasei;	CCUG 27320^T; -	+++	−/+
L. lactis 53	CCUG 27320^T; -	+++	−/+
L. rhamnosus;	ATCC 7469^T; CECT 410^T	++++	-
E. faecium	ATCC 7469^T; CECT 410^T	++++	-
L. reuteri;	NCFB 2656^T;	+++	-
E. coli O157:H7	NCTC 12900^T	+++	-
S. aureus;	CECT 976^T; -	-	++++
G. vaginalis 51	CECT 976^T; -	-	++++
Shigella;	ATCC 12022^T; -	-	++++
G. vaginalis 101	ATCC 12022^T; -	-	++++
L. seeligeri;	CECT 917^T; -	-	++++
G. vaginalis AMD	CECT 917^T; -	-	++++
E. aerogenes;	CECT 684^T; -	-	++++
G. vaginalis ATCC	CECT 684^T; -	-	++++
L. pentosus;	CECT 4023^T;	++++	++++
G. vaginalis ATCC;	-;
E. faecalis	CECT 184^T
L. casei;	CECT 5275^T;	++++	++++
G. vaginalis AMD;	-;
A. vaginae	CCUG 38953^T
L. rhamnosus;	CECT 288^T;	++++	++++
G. vaginalis 101;	-;
A. vaginae	CCUG 42099^T
L. crispatus;	ATCC 33820^T;	++++	++++
G. vaginalis 51;	-;
A. vaginae	CCUG 44116^T
L. casei;	CECT 5275^T;	++++	-
L. mesenteroides;	-;
A. vaginae	CCUG 38953^T

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; (+) Poor hybridization; (+++) Good hybridization; (++++) Optimal hybridization. Median values from the three experiments for each strain are shown in the table.

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