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Characterization of the brewery spoilage bacterium Obesumbacterium proteus by automated ribotyping and development of PCR methods for its biotype 1
Author(s) -
Koivula T.T.,
Juvonen R.,
Haikara A.,
Suihko M.L.
Publication year - 2006
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2005.02794.x
Subject(s) - ribotyping , library science , christian ministry , clinical microbiology , microbiology and biotechnology , biology , polymerase chain reaction , political science , law , computer science , gene , genetics
Aims: To study the ability of automated ribotyping to characterize Obesumbacterium proteus and Hafnia alvei , to design primers and to evaluate standard end‐point and real‐time PCR for the detection of O. proteus biotype 1 in beer and in brewers's yeast‐containing samples. Methods and Results: Automated ribotyping was carried out using the standard method with Eco RI and Pvu II. The digestions with both enzymes clearly differentiated O. proteus biotypes 1 and 2 and H. alvei . PCR primers were designed according to the 16S rRNA gene sequence of the O. proteus type strain. Two primer sets (Obs137–Obs558 and Obs137–Obs617) detected O. proteus biotype 1 and H. alvei but not O. proteus biotype 2 or other tested beer spoilage bacteria (40 species) in the end‐point and real‐time PCR, indicating their high specificity. The detection limit for O. proteus was 160–1600 CFU 100 ml −1 beer in the end‐point PCR reactions and ≤160 CFU 100 ml −1 beer in the real‐time PCR reactions. More cells (from 16 to 3200) were needed for detection in the presence of brewer's yeast cells. Conclusions: Automated ribotyping is a useful tool to characterize and identify O. proteus and H. alvei isolates. The designed primers are suitable for the rapid detection of O. proteus biotype 1 and H. alvei in brewery samples by PCR. Significance and Impact of the Study: Automated ribotyping and PCR could improve microbiological quality control in breweries by facilitating the detection, identification and tracing of spoilage bacteria.