Development of Method for Evaluating Safety of Water Applying PCR

PCR评价水安全性方法的研制

• 批准号: 05453200
• 负责人: OHGAKI Shinichiro
• 金额: $ 4.54万
• 依托单位: THE UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05453200/
• 关键词: PCR; RNA phage; MPN; Indicator for viruses; Ultraviolet Disinfection; Monitoring

PCR method was developed as a quantitative detection of viruses in water, including the investigation of pre-treatment method. Application of PCR method to the water treated by a treatment process (Ultraviolet Irradiation). RNA phage Q-beta was used for the experiment as a model virus.In order to inprove the quantitatative PCR,optimisation of beffer concentration in both reverse transcription and DNA amplify, and development of nested PCR were done. Result showed the detection limit was 1 PFU.It was suggested that using computer program to calculate MPN lead to the utilization of quantitative PCR date.Effect of ultraviolet disinfection on the PCR detection was also investigated. Value of surviving phage concentration measured by conventional plapue method and that measured by PCR method were compared. Result showed that PCR measurment detected most of inactivated phages. Quantity of detection was assumed to depend on the ratio of the length of target RNA of PCR detection to the whole lengh of phage RNA.These results showed that PCR method was hard to be applied to the monitoring the performance of ultraviolet disinfection. Further research was required to utilize the PCR to measure the safety of water.

建立了一种定量检测水中病毒的PCR方法，包括前处理方法的研究。PCR方法在处理工艺（紫外线照射）处理过的水中的应用。以噬菌体Q-β为模型病毒，优化了反转录和DNA扩增的缓冲液浓度，建立了巢式PCR方法。结果表明，该方法的检出限为1 PFU，建议利用计算机程序计算MPN，以实现定量PCR数据的利用，并探讨了紫外线消毒对PCR检测的影响。比较了常规平板法和PCR法测定噬菌体存活浓度的结果。结果表明，PCR检测可检出绝大多数灭活大肠杆菌。实验结果表明，PCR检测的定量依赖于PCR检测的靶RNA长度与噬菌体RNA全长的比值，PCR方法难以用于紫外线消毒效果的监测。需要进一步研究利用PCR来测量水的安全性。

项目成果

Hiroyuki Katayama, Naoyuki Kamiko , Shinichiro Ohgaki: "Quantitative measurement of RNA coliphage Q beta by using PCR method" Proc. of Annual Conf. Jap. Soc. of Water Environ.29. (1995)

Hiroyuki Katayama、Naoyuki Kamiko、Shinichiro Ohgaki：“使用 PCR 方法定量测量 RNA 大肠杆菌噬菌体 Q beta”Proc。

片山浩之,神子直之,大垣眞一郎: "水中のRNAファージのポリメラーゼチェインリアクション(PCR)法による検出" 土木学会第48回年次学術講演会講演概要集第2部. 1246-1247 (1993)

Hiroyuki Katayama、Naoyuki Miko、Shinichiro Ogaki：“通过聚合酶链反应（PCR）法检测水中的RNA噬菌体”日本土木工程学会第48届年度学术会议论文集，第2部分.1246-1247（1993）

L.Sunun: "Application of Polymerase ChainReaction to Detect RNAColiphage Qβ inEnrironmental Water Samples)" Water Science and Technology. (1995)

L. Sunun：“应用聚合酶链式反应检测环境水样中的 RNA 大肠杆菌噬菌体 Qβ”，水科学与技术 (1995)。

片山浩之,大垣眞一郎,神子直之: "紫外線によって不活化されたファージのPCR法による検出" 第28回日本水環境学会年会講演集. 494-495 (1994)

Hiroyuki Katayama、Shinichiro Ogaki、Naoyuki Miko：“通过PCR法检测紫外线灭活的噬菌体”日本水环境学会第28届年会记录494-495（1994）。

片山浩之.神子直之.大垣眞一郎: "紫外線によって不活化されたファージのPCR法による検出" 日本水環境学会年会講演集. 第28回(発表予定). (1994)

Hiroyuki Katayama，Naoyuki Ogaki：“通过 PCR 方法检测紫外线灭活的噬菌体”，日本水环境学会年会记录（1994 年）。