Detection and identification of Pectinatus brewery contaminants based on the gene for the major outer membrane protein

Considering the recurrence of Pectinatus in breweries, detection and differentiation of contaminating isolates is important for quality control to mitigate recurring or newly developed spoilage problems. As a potential new PCR target, the gene for a major outer membrane protein (OMP) was sequenced for three species of Pectinatus, including one Pectinatus haikarae, two P. cerevisiiphilus, and three P. frisingensis isolates. Conserved regions in the OMP gene enabled the design of a multiplex PCR that concurrently targets all three species. The multiplex PCR demonstrates specificity for brewery Pectinatus isolates, and does not amplify DNA from various Megasphaera, Selenomonas, or Zymophilus species. Additionally, the OMP gene contains variable regions that allow for differentiation of Pectinatus strains. As such, by sequencing individual PCR amplicons, it is possible to determine if the Pectinatus contaminant is new or recurring. Furthermore, by targeting the OMP gene, strain-specific Pectinatus detection methods can be designed to enable rapid monitoring and quality control of recurring Pectinatus contaminants within a given brewery. © 2014 American Society of Brewing Chemists, Inc