Timely approaches to identify probiotic species of the genus Lactobacillus

Table 1 Experiment duration and detection level for each method

Method	Duration (h^a)	Time in total (h)		Detection at the level of…			Quantification
		Culture-dependent	Culture-independent^b	Genus	Species	Subspecies
Morphology	*^c	~ 48	^{/ d}	-^e	-	-	-
FTIR	* + 1 h (analyzation)	~ 49	^/	+^f	+	-	-
MALDI-TOF MS	* + 1 h (analyzation)	~ 49	^/	+	+	+	-
API 50 CHL	* + 48 h (incubation)	~ 96	^/	+	+	-	-
BIOLOG	* + 25 h (24 h incubation of AN MicroPlate^TM + 1 h analyzation)	~ 72	^/	+	+	-	-
16S/23S rRNA PCR + sequencing	* + 31 h (4 h DNA isolation + 3 h PCR + 24 h sequencing)	~ 79	~ 31	+	+	-	-
PCR-DGGE	* + 7 h (4 h DNA isolation + 3 h PCR + electrophoresis)	~ 55	~ 7	+	+	-	(+)^g
RAPD	* + 7 h (4 h DNA isolation + 3 h PCR + electrophoresis)	~ 55	~ 7	+	+	+	-
SSCP	* + 7 h (4 h DNA isolation + 3 h PCR + electrophoresis)	~ 55	~ 7	+	+	-	-
MLST	* + 31 h (4 h DNA isolation + 3 h qPCR + 24 h sequencing)	~ 79	~ 31	+	+	+	-
qPCR	* + 7 h (4 h DNA isolation + 3 h qPCR)	~ 55	~ 7	+	+	-	+
SSR	* + 7 h (4 h DNA isolation + 3 h PCR + electrophoresis)	~ 55	~ 7	+	+	+	-
WGS	* + 4 h DNA isolation + 36 h Sequencing, annotation, etc.)	~ 88	~ 40	+	+	+	-

^a h, hour(s).
^b duration exclusive cultivation of the strains (48 h).
^c *, isolation of the strains by plating on different media (MRS broth [Roth, Germany [14]], LBS agar [Becton Dickinson, USA [15]], COL and CHOC plates [Sarstedt, Germany]).
^d /, identification of the strains not possible by working culture-independent.
^e -, detection at the level of… is not possible.
^f +, detection at the level of… is possible.
^g (+), a limited quantification is possible using PCR-DGGE.

ISSN: 1757-4749