Detection of beer spoilage bacteria Megasphaera and Pectinatus by polymerase chain reaction and colorimetric microplane hybridization

Reetta Satokari (Corresponding Author), Riikka Juvonen, Kirstie Mallison, Atte von Wright, Auli Haikara

Research output: Contribution to journalArticleScientificpeer-review

42 Citations (Scopus)

Abstract

Anaerobic bacteria of the genera Megasphaera and Pectinatus cause beer spoilage by producing off flavours and turbidity.
Detection of these organisms is complicated by the strict anaerobic conditions and lengthy incubation times required for their cultivation, consequently there is a need for more rapid detection methods.
A polymerase chain reaction (PCR) method and a colorimetric microplate hybridization assay were developed for the rapid and specific detection of Megasphaera cerevisiae and Pectinatus spp. A biotinylated primer pair was designed for the amplification of a 403 base pair (bp) fragment of the M. cerevisiae 16S rRNA gene and a primer pair from literature was used for the amplification of an 816 bp fragment of Pectinatus 16S rRNA gene. Amplified PCR products were analyzed by the colorimetric microplate hybridization method in which a biotinylated PCR product was captured by streptavidin and hybridized with a digoxigenin-labelled oligonucleotide probe.
In the final step an enzyme-linked antibody and a colorimetric reaction were utilized. A simple and rapid sample treatment was set up for the PCR detection of contaminants in beer. Detection of M. cerevisiae (≥5·103 colony forming units [cfu]/100 ml) and Pectinatus frisingensis (≥5·105 cfu/100 ml) in beer was successful, but the sensitivity of the assay still needs to be improved for direct detection of the small amounts of bacteria present in beer.
Original languageEnglish
Pages (from-to)119-127
JournalInternational Journal of Food Microbiology
Volume45
Issue number2
DOIs
Publication statusPublished - 1998
MoE publication typeA1 Journal article-refereed

Fingerprint

Pectinatus
Megasphaera
Megasphaera cerevisiae
hybridization
polymerase chain reaction
beers
Bacteria
Polymerase Chain Reaction
rRNA Genes
Base Pairing
Stem Cells
Digoxigenin
Pectinatus frisingensis
Streptavidin
Anaerobic Bacteria
Oligonucleotide Probes
ribosomal RNA
streptavidin
digoxigenin
oligonucleotide probes

Keywords

  • Megasphaera
  • Pectinatus
  • Beer spoilage
  • PCR
  • Colorimetric microplate hybridization

Cite this

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title = "Detection of beer spoilage bacteria Megasphaera and Pectinatus by polymerase chain reaction and colorimetric microplane hybridization",
abstract = "Anaerobic bacteria of the genera Megasphaera and Pectinatus cause beer spoilage by producing off flavours and turbidity. Detection of these organisms is complicated by the strict anaerobic conditions and lengthy incubation times required for their cultivation, consequently there is a need for more rapid detection methods. A polymerase chain reaction (PCR) method and a colorimetric microplate hybridization assay were developed for the rapid and specific detection of Megasphaera cerevisiae and Pectinatus spp. A biotinylated primer pair was designed for the amplification of a 403 base pair (bp) fragment of the M. cerevisiae 16S rRNA gene and a primer pair from literature was used for the amplification of an 816 bp fragment of Pectinatus 16S rRNA gene. Amplified PCR products were analyzed by the colorimetric microplate hybridization method in which a biotinylated PCR product was captured by streptavidin and hybridized with a digoxigenin-labelled oligonucleotide probe. In the final step an enzyme-linked antibody and a colorimetric reaction were utilized. A simple and rapid sample treatment was set up for the PCR detection of contaminants in beer. Detection of M. cerevisiae (≥5·103 colony forming units [cfu]/100 ml) and Pectinatus frisingensis (≥5·105 cfu/100 ml) in beer was successful, but the sensitivity of the assay still needs to be improved for direct detection of the small amounts of bacteria present in beer.",
keywords = "Megasphaera, Pectinatus, Beer spoilage, PCR, Colorimetric microplate hybridization",
author = "Reetta Satokari and Riikka Juvonen and Kirstie Mallison and Wright, {Atte von} and Auli Haikara",
year = "1998",
doi = "10.1016/S0168-1605(98)00154-8",
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volume = "45",
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Detection of beer spoilage bacteria Megasphaera and Pectinatus by polymerase chain reaction and colorimetric microplane hybridization. / Satokari, Reetta (Corresponding Author); Juvonen, Riikka; Mallison, Kirstie; Wright, Atte von; Haikara, Auli.

In: International Journal of Food Microbiology, Vol. 45, No. 2, 1998, p. 119-127.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Detection of beer spoilage bacteria Megasphaera and Pectinatus by polymerase chain reaction and colorimetric microplane hybridization

AU - Satokari, Reetta

AU - Juvonen, Riikka

AU - Mallison, Kirstie

AU - Wright, Atte von

AU - Haikara, Auli

PY - 1998

Y1 - 1998

N2 - Anaerobic bacteria of the genera Megasphaera and Pectinatus cause beer spoilage by producing off flavours and turbidity. Detection of these organisms is complicated by the strict anaerobic conditions and lengthy incubation times required for their cultivation, consequently there is a need for more rapid detection methods. A polymerase chain reaction (PCR) method and a colorimetric microplate hybridization assay were developed for the rapid and specific detection of Megasphaera cerevisiae and Pectinatus spp. A biotinylated primer pair was designed for the amplification of a 403 base pair (bp) fragment of the M. cerevisiae 16S rRNA gene and a primer pair from literature was used for the amplification of an 816 bp fragment of Pectinatus 16S rRNA gene. Amplified PCR products were analyzed by the colorimetric microplate hybridization method in which a biotinylated PCR product was captured by streptavidin and hybridized with a digoxigenin-labelled oligonucleotide probe. In the final step an enzyme-linked antibody and a colorimetric reaction were utilized. A simple and rapid sample treatment was set up for the PCR detection of contaminants in beer. Detection of M. cerevisiae (≥5·103 colony forming units [cfu]/100 ml) and Pectinatus frisingensis (≥5·105 cfu/100 ml) in beer was successful, but the sensitivity of the assay still needs to be improved for direct detection of the small amounts of bacteria present in beer.

AB - Anaerobic bacteria of the genera Megasphaera and Pectinatus cause beer spoilage by producing off flavours and turbidity. Detection of these organisms is complicated by the strict anaerobic conditions and lengthy incubation times required for their cultivation, consequently there is a need for more rapid detection methods. A polymerase chain reaction (PCR) method and a colorimetric microplate hybridization assay were developed for the rapid and specific detection of Megasphaera cerevisiae and Pectinatus spp. A biotinylated primer pair was designed for the amplification of a 403 base pair (bp) fragment of the M. cerevisiae 16S rRNA gene and a primer pair from literature was used for the amplification of an 816 bp fragment of Pectinatus 16S rRNA gene. Amplified PCR products were analyzed by the colorimetric microplate hybridization method in which a biotinylated PCR product was captured by streptavidin and hybridized with a digoxigenin-labelled oligonucleotide probe. In the final step an enzyme-linked antibody and a colorimetric reaction were utilized. A simple and rapid sample treatment was set up for the PCR detection of contaminants in beer. Detection of M. cerevisiae (≥5·103 colony forming units [cfu]/100 ml) and Pectinatus frisingensis (≥5·105 cfu/100 ml) in beer was successful, but the sensitivity of the assay still needs to be improved for direct detection of the small amounts of bacteria present in beer.

KW - Megasphaera

KW - Pectinatus

KW - Beer spoilage

KW - PCR

KW - Colorimetric microplate hybridization

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DO - 10.1016/S0168-1605(98)00154-8

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EP - 127

JO - International Journal of Food Microbiology

JF - International Journal of Food Microbiology

SN - 0168-1605

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ER -