Diversity analysis of beer spoiling gram-negative isolates using PCR fingerprinting and computer- assisted analysis

摘要

Seventy-two Pectinatus strains, isolated from ten brewery environments throughout the world or sourced from culture collections, were analysed by three types of polymerase chain reaction (PCR) fingerprinting. This analysis was conducted in order to study diversity and relatedness within each genus and to compare sources within each brewery and between different breweries. All strains were subjected to repetitive-sequence-based (rep) PCR profiling using three primer sets: (GTG)_5 BOX and REP. Fingerprint profiles were obtained from field and reference strains of Pectinatus frisingensis, using all three primer sets. Pectinatus cerevisiiphilus isolates yielded (GTG)_5 - and BOX-PCR fingerprint profiles only. Cluster analysis of combined (GTG)_5, BOX and REP fingerprint profiles revealed several groups. A total of eleven genomic fingerprint types were generated within the collection of Pectinatus isolates, which included the reference strains and 61 field isolates. Rep-PCR genomic fingerprinting combines high resolution with high reproducibility and is rapid and inexpensive, compared to amplified fragment length polymorphism (AFLP) analysis and automated Ribotyping. Rep-PCR genomic fingerprinting allows for database mediated typing, identification and strain specific tracking. The results are valuable for tracking and tracing microorganisms in the case of spoilage and it can also be applied to profile other brewery related bacteria and yeast.