Screening of Beer-Spoilage Bacteria Using,,,the LightCycler PCR Workflow System
To gain information about sensitivity and specificity ofthe LightCycler foodproof Beer Screening Kit eight differentbeer-spoilage bacteria species and one mixture oftwo different species of Bitburger reference stock weretested: L. brevis P. frisingensis Megasphaera l
To gain information about sensitivity and specificity of
the LightCycler foodproof Beer Screening Kit, eight different
beer-spoilage bacteria species and one mixture of
two different species of Bitburger reference stock were
tested:
L. brevis
P. frisingensis
Megasphaera
P. cerevisiiphilus
L. lindneri
L. collinoides
Pediococcus
L. parabuchneri
mix of Pediococcus and L. parabuchneri
The samples were tested in four dilution steps: 106 colony
forming units (cfu)/ml, 104 cfu/ml, 102 cfu/ml, and
101 cfu/ml. The samples were prepared as duplicates.
Melting-curve analysis
20 different DNA samples of single and mixed beerspoilage
bacteria were analyzed. Sample preparation was
not necessary in this case. DNA concentrations were different
for each sample, ranging between 5 copies and
3,000 copies. The different beer-spoilage bacteria were
analyzed by melting-curve analysis.
Results and Discussion
Suitability testing of routine samples
Ten of 58 samples showed positive results that were
clearly identified as beer-spoilage bacteria via meltingcurve
analysis. The remaining 48 samples tested negative,
and the internal control (IC) was amplified with a
similar PCR efficiency. These results indicate that the
MagNA Pure LC efficiently removed potential PCR
inhibitors originating either from the beer routine sample
or from the enrichment culture (data not shown). This
clearly demonstrates the suitability and the reliability of
the '"/>
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