Rapid On-chip flow Cytometric Detection of Listeria monocytogenes in Milk
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- Ikeda Masafumi
- Environmental Science and Microbiology, Graduate School of Pharmaceutical Sciences, Osaka University
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- Yamaguchi Nobuyasu
- Environmental Science and Microbiology, Graduate School of Pharmaceutical Sciences, Osaka University
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- Nasu Masao
- Environmental Science and Microbiology, Graduate School of Pharmaceutical Sciences, Osaka University
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Abstract
Listeria monocytogenes (L. monocytogenes) is a Gram-positive, intracellular bacterium that can cause severe infections in humans. Contamination by L. monocytogenes in foods represents a potential public health problem. Here we describe a rapid method for the detection of L. monocytogenes in milk by flow cytometry based on microfluidics (on-chip flow cytometry). This rapid identification of L. monocytogenes is based on fluorescence in situ hybridization (FISH) with a Cy5-labeled rRNA-targeting oligonucleotide probe. FISH experiments were successfully analyzed using a commercially available on-chip flow cytometry. FISH results with bacterial cultures indicated that the L. monocytogenes probe RL-2 was hybridized with 4 major strains of L. monocytogenes (serotype 1/2a, 1/2b, 1/2c and 4b) but not with other Listeria strains or milk spoilage bacteria. Specific FISH detection of L. monocytogenes was accomplished using the probe RL-2 when the milk contained a mixture of other bacterial species. The positive identification of L. monocytogenes in milk was completed within 5 hr (milk clearing: 40 min, hybridization: 3.5 hr, on-chip analysis: 30 min). The method presented in this study allows the specific and rapid identification of L. monocytogenes in milk.
Journal
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- Journal of Health Science
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Journal of Health Science 55 (5), 851-856, 2009
The Pharmaceutical Society of Japan
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Details 詳細情報について
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- CRID
- 1390001204497362944
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- NII Article ID
- 130000127978
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- NII Book ID
- AA11316464
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- ISSN
- 13475207
- 13449702
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- NDL BIB ID
- 10376576
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- CiNii Articles
- KAKEN
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- Abstract License Flag
- Disallowed