( To gai...)Screening of Beer-Spoilage Bacteria Using,,,the LightCycler PCR Workflow System

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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