2015 Fiscal Year Final Research Report
Differentiation of dead cells and live cells in food by using the propidium monoazide
| Project/Area Number |
25450303
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Aquatic life science
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| Research Institution | Tokyo University of Marine Science and Technology |
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Principal Investigator |
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Keywords | 食品微生物 / 微生物定量 / 生菌死菌判別 / リアルタイムPCR |
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| Outline of Final Research Achievements |
PCR has been widely used for detection or enumeration of various bacterial cells in various samples, including food samples, due to its rapidity, simplicity and specificity. However, one of the underlying problems with PCR is the lack of differentiation between viable and non-viable cells, which can lead to false-positive results and overestimation. Recently, several researchers have studied the differentiation of DNA molecules derived from non-viable cells by photo-induced inactivation of DNA using propidium monoazide (PMA). However, the protocols for PMA treatment have yet to be established. In this study, we improved the light exposure apparatus using LED to resolve this issue. Various LED elements were arrayed in an apparatus for PMA treatment in order to suppress false-positive PCR results from DNA derived from dead bacterial cells. LEDs emitting 375 nm were found to facilitate PMA-DNA interaction, thus leading to efficient inhibition of PCR of DNA derived from dead cells.
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| Free Research Field |
食品微生物学
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