Characterization of the brewery spoilage bacterium Obesumbacterium proteus by automated ribotyping and development of PCR methods for its biotype 1

Teija Koivula, Riikka Juvonen, Auli Haikara, Maija-Liisa Suihko (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

21 Citations (Scopus)

Abstract

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 EcoRI and PvuII. 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.
Original languageEnglish
Pages (from-to)398-406
Number of pages9
JournalJournal of Applied Microbiology
Volume100
Issue number2
DOIs
Publication statusPublished - 2006
MoE publication typeA1 Journal article-refereed

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Ribotyping
Proteus
Hafnia alvei
Bacteria
Polymerase Chain Reaction
rRNA Genes
Quality Control
Saccharomyces cerevisiae
Limit of Detection
Digestion

Keywords

  • automated ribotyping
  • biotype 1
  • Obesumbacterium proteus
  • real-time PCR

Cite this

@article{ca7a2b8fdb6a44a08a10e8d2c22f5b9e,
title = "Characterization of the brewery spoilage bacterium Obesumbacterium proteus by automated ribotyping and development of PCR methods for its biotype 1",
abstract = "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 EcoRI and PvuII. 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.",
keywords = "automated ribotyping, biotype 1, Obesumbacterium proteus, real-time PCR",
author = "Teija Koivula and Riikka Juvonen and Auli Haikara and Maija-Liisa Suihko",
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doi = "10.1111/j.1365-2672.2005.02794.x",
language = "English",
volume = "100",
pages = "398--406",
journal = "Journal of Applied Microbiology",
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Characterization of the brewery spoilage bacterium Obesumbacterium proteus by automated ribotyping and development of PCR methods for its biotype 1. / Koivula, Teija; Juvonen, Riikka; Haikara, Auli; Suihko, Maija-Liisa (Corresponding Author).

In: Journal of Applied Microbiology, Vol. 100, No. 2, 2006, p. 398-406.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

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AU - Koivula, Teija

AU - Juvonen, Riikka

AU - Haikara, Auli

AU - Suihko, Maija-Liisa

PY - 2006

Y1 - 2006

N2 - 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 EcoRI and PvuII. 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.

AB - 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 EcoRI and PvuII. 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.

KW - automated ribotyping

KW - biotype 1

KW - Obesumbacterium proteus

KW - real-time PCR

U2 - 10.1111/j.1365-2672.2005.02794.x

DO - 10.1111/j.1365-2672.2005.02794.x

M3 - Article

VL - 100

SP - 398

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JO - Journal of Applied Microbiology

JF - Journal of Applied Microbiology

SN - 1364-5072

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