Discrimination of Viable and Dead Escherichia coli Cells by Quantitative PCR with Propidium Monoazide
| Project/Area Number |
19560555
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Civil and environmental engineering
|
|---|
| Research Institution | Hachinohe National College of Technology |
|---|
Principal Investigator |
YAGUCHI Jyunichi Hachinohe National College of Technology, 建設環境工学科, 教授 (80342450)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
FUKUSHI Kensuke 東京大学, サスティナビリティ連携研究機構, 准教授 (30282114)
金子 仲一郎 独立行政法人国立高等専門学校機構八戸工業高等専門学校, 建設環境工学科, 助手 (70099761)
|
|---|
| Research Collaborator |
SAWAYA Keisuke 八戸工業高等専門学校, 専攻科・建設環境工学専攻, 学生
YOKOMACHI Keisuke 八戸工業高等専門学校, 専攻科・建設環境工学専攻, 学生
YOSHIDA Yoshitugu 八戸工業高等専門学校, 建設環境工学科, 学生
KUDO Eiichi 八戸工業高等専門学校, 建設環境工学科, 学生
NAKAYAMA Yusuke 八戸工業高等専門学校, 建設環境工学科, 学生
RUIKE Syo 八戸工業高等専門学校, 建設環境工学科, 学生
|
|---|
| Project Period (FY) |
2007 – 2009
|
| Project Status |
Completed (Fiscal Year 2009)
|
| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2009: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2008: ¥390,000 (Direct Cost: ¥300,000、Indirect Cost: ¥90,000)
Fiscal Year 2007: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
|
|---|
| Keywords | 水質 / リアルタイムPCR / DNA / 大腸菌 / 閾値サイクル数 / 熱処理 / PMA / サイクル数 / 生菌 / 塩素消毒 / 不活性化速度定数 / 生理的活性 |
|---|
| Research Abstract |
A real-time PCR method was applied to the detection of fecal indicator, Escherichia coli based on the uid A gene sequence. Rapid and reliable quantitative estimation of E.coli was conducted in concentration ranging from 1×10^1 to 1×10^6 cfu/tube. A combination of PMA (propidium monoazide) and real-time PCR was developed to enumerate viable E.coli in the presence of dead cells. Mixtures of viable cells and heat-treated cells were subjected to real-time PCR with PMA treatment. Viable cell counts were linearly related to real-time PCR threshold cycle values for PMA-treated cells as long as the ratio of dead cells to viable cells was no greater than 1×10^1. The effect of chlorine on the physiological viability of bacteria in the river was alsoinvestigated. Microscopic viable bacteria were more chlorine resistant than culturablebacteria. The inactivation rate coefficients of direct viable bacteria were one-secondto third those of culturable bacteria.
|
Report
(4 results)
Research Products
(11 results)
