EFFECTS OF FILTRATION FOR ENUMERATION OF VIABLE <i>ESCHERICHIA COLI</i> BY QUANTITATIVE PCR AFTER PROPIDIUM MONOAZIDE TREATMENT WITH A LED LAMP
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- RUIKE Wataru
- 東北大学大学院 工学研究科
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- YAMAMOTO Ayumu
- 八戸工業高等専門学校 産業システム工学科
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- YAGUCHI Junichi
- 八戸工業高等専門学校 産業システム工学科
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- KUBOTA Kengo
- 東北大学大学院 工学研究科
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- LI Yu-You
- 東北大学大学院 工学研究科
Bibliographic Information
- Other Title
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- PMA-qPCR法へのLED光源の適用とろ過濃縮の影響
- PMA-qPCRホウ エ ノ LED コウゲン ノ テキヨウ ト ロカ ノウシュク ノ エイキョウ
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Abstract
Quantitative detection of health-related microorganisms is of importance for risk assessment of environmental context, but quantitative real-time PCR (qPCR) alone meets a difficulty to distinguish viable and dead cells. Propidium monoazide (PMA) treatment followed by qPCR has been introduced to overcome this issue. In this study, a LED lamp, instead of a halogen lamp, was used for cross-linking of PMA and DNA, and the effects of filtration for the enumeration of viable Escherichia coli by PMA-qPCR was investigated. PMA-qPCR was applicable to suspended cells after the treatment with 100μM PMA followed by 15 min exposure to a LED lamp. On the other hand, PMA-qPCR was not able to distinguish viable and dead cells after filtration. Cell treatment with LIVE/ DEAD BacLight Bacterial Viability Kits indicated that the most of cell after a filtration procedure were damaged, suggesting the filtration procedure is one crucial point for PMA-qPCR of filtered samples.
Journal
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- Journal of Japan Society of Civil Engineers, Ser. G (Environmental Research)
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Journal of Japan Society of Civil Engineers, Ser. G (Environmental Research) 72 (7), III_315-III_323, 2016
Japan Society of Civil Engineers