Peter Trabold (1),
Daniela Bocioaga (1), I. Cristina McGuire (1), Stephanie Morse (1),
Amanda Ruby (1), Gwendolyn Spizz (1); (1) Rheonix Corporation, Grand
Island, NY, U.S.A.
Finishing and Stability
Poster
Despite the inhospitable environment of beer toward the growth of
microorganisms, craft brews remain susceptible to various spoilage
organisms that have developed mechanisms that enable growth in the
presence of hops, alcohol, and acidic conditions. The standard detection
strategy for these spoilage organisms has traditionally involved
culturing, a process that may take up to a week to obtain results. This
wait is problematic for brewers and may result in delayed release or
release of untested and potentially spoiled product to market. In
contrast to culture, molecular detection provides a rapid, sensitive,
and specific method of identifying the presence of beer spoilage
organisms. Available kits on the market identify many species of
spoilers, regardless of whether the specific strain contains genes
associated with spoilage, or focus on a limited number of beer spoilage
genes, risking lack of detection of other relevant spoilers. Most
available tests require pre-enrichment and other lengthy preparative
steps, in addition to sophisticated molecular lab equipment that may not
be present in a craft brewery. This study describes the development of
the Beer SpoilerAlert assay, a robust sample-to-results molecular
detection system for beer spoilage organisms, including various species
of lactic acid bacteria (LAB), and four genes associated with beer
spoilage in LAB. Concurrently, the assay detects the presence of the
wild yeast Brettanmoymces bruxellenis and brewer’s yeast.
Samples taken at any point during the brewing process are loaded into
sample tubes and placed in the sample rack. The sample rack, cartridges,
and reagent kit are placed in the Encompass Optimum workstation and all
processes required for lysing organisms, extracting nucleic acids,
amplifying and detecting target genes, and analyzing results, are
automatically performed without user intervention on the Rheonix CARD
(Chemistry and Reagent Device) cartridges in the workstation. Reagents
are dispensed by an onboard robot and liquid is moved via microfluidic
pumps and channels within the cartridge. Amplification occurs via the
onboard thermocycler and endpoint detection occurs through hybridization
to a low-density capture array. Captured targets are detected and
analyzed by an onboard camera and imaging software, which provides the
user with a report of which genes and/or organisms are present. Four
individual samples are analyzed per cartridge, with six cartridges per
run, resulting in twenty-four independent samples analyzed in 5 hr, with
minimal hands-on time. Due to the sensitivity of the assay, a
pre-enrichment step is not required in most instances. However, if the
user prefers to do so, a pre-enrichment step is compatible with this
assay with no further modifications. The results described demonstrate
validation of the assay using beer samples spiked with known microbes
and actual beer samples suspected or known to contain spoilage
organisms. Results were verified through conventional culturing methods.
Peter Trabold received a B.A. degree in biology and philosophy and
an MBA and a Ph.D. in molecular and cellular biology from the
University at Buffalo. He began working for ZeptoMetrix Corporation in
2005 and established the Bacteriology Department in 2007. As the manager
of this department, he was responsible for the initial development of
four distinct product lines for the research and clinical diagnostics
infectious disease markets. In 2012, Peter was promoted to director of
business development at ZeptoMetrix. Currently, Peter is the director of
business development at Rheonix Corporation. In addition, Peter is
currently president of the Western New York branch of ASM.
