| 环境水体中肠道病原细菌的定量PCR检测 |
| 摘要点击 2321 全文点击 1778 投稿时间：2007-05-21 修订日期：2007-10-15 |
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| 中文关键词 地表水 通用引物 病原细菌 定量PCR 膜过滤 |
| 英文关键词 surface water universal primer enteric pathogenic bacteria quantitative polymerase chain reaction(QPCR) membrane filtration (MF) |
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| 中文摘要 |
| 采用通用引物，通过实时荧光定量PCR方法(QPCR)，对西安市5处地表水体中肠道病原细菌的细胞密度进行4个月的连续检测，并将QPCR检测结果与滤膜法测得的大肠菌群CFU值进行比较分析.结果显示， QPCR法可信度为94%，最小检测值为每管未稀释DNA提取物含2.7个大肠杆菌细胞.5个水体（N=60）检测结果表明，QPCR检测结果是大肠菌群CFU的2.2～5倍.病原细菌的几何平均值范围，QPCR法在25～67?000 CCE/100 mL之间，MF法大肠菌群为3～45?000 CFU/100 mL之间.2种方法的离散和回归分析表明，QPCR检测结果与大肠菌群CFU显著正相关，秩相关系数为R=0.983. |
| 英文摘要 |
| A rapid quantitative polymerase chain reaction (QPCR) analysis method with universal primers was developed to detect cell densities of the enteric pathogenic bacteria from 5 surface water of Xi'an City for 4 months continuously. And the detection results by QPCR method were compared with counts of coliforms colony-forming units (CFU) determined by membrane filter (MF) analysis. The results showed that QPCR method had an estimated 94% confidence, and detection limit was 2.7 Escherichia coli cells per sample in undiluted DNA extracts. For five surface waters(N=60), the geometric mean of pathogenic bacteria concentration determined by QPCR was 2.2-5 times of corresponding coliform CFU determined by MF analysis. Using QPCR analysis, these geometric means of pathogenic bacteria concentration ranged from 25 CCE/100 mL to 67?000 CCE/100 mL. Using MF culture analysis, coliforms ranged from 3 CFU/100 mL to 45?000 CFU/100 mL. Regression analysis showed that there was a significant positive correlation between pathogenic bacteria determined by QPCR method and coliforms determined by MF method, the correlation coefficient (R) was 0.983. |
