Establishment of a digital PCR-based genotyping method for noroviruses in water

2016 Fiscal Year Final Research Report

• Project/Area Number: 26820222
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Civil and environmental engineering
• Research Institution: Kyoto University
• Principal Investigator: Hata Akihiko 京都大学, 工学研究科, 特別研究員(PD) (70726306)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 健康関連微生物 / ノロウイルス / Real-time PCR / デジタルPCR / 水系感染症

Outline of Final Research Achievements

It is known that genetically broad types of noroviruses are exist together in the water environment. Quantification of single genotype of noroviruses seems to be important to understand characteristics of noroviruses, in view of infection risk via water and epidemiology, more deeply. The aim of the study was to establish a method for simultaneous quantification of “whole norovirus” and “a genotype of norovirus” using the novel digital PCR platform.
Water samples in Japan were continuously collected and analyzed to determine epidemic norovirus genotypes. Based on nucleotide sequences of the epidemic types and those on the database, fluorescent probes, which react to each epidemic genotype specifically, were designed. In the conventional real-time PCR platform, the fluorescent probes worked ideally, while they resulted in non-specific signal in the digital PCR platform. The results imply that further improvement is necessary for establishing the assay.

Free Research Field

水環境学