導電性平膜と遺伝子工学的手法を用いたレジオネラ菌検出とその検出感度に関する検討

概要

論文の詳細を見る
Identification of Legionella is requiring more than a week. A method for rapid detection is therefore required to restrict the increasing range of Legionnaires' disease and to control Legionella. While the nucleic acid amplification test (NAT) has been applied to bacterial screening, it has not yet been applied to the detection of the few Legionella commonly found in environmental waters like hot springs given the relative limits of the sample volumes that can be processed using NAT. We therefore designed a genetic detection system using a metal-coated flat sheet (MCFS) membrane for Legionella that operates as follows : The MCFS-membrane is coated with silver on a flat filtration membrane. Legionella become concentrated on the MCFS-membrane when water from a hot spring is aspirated through the membrane. The genes of Legionella that become concentrated in the membrane are then isolated by applying an electric field. Legionella can then identify by the polymerase chain reaction (PCR) using specific primers. Here we report the first attempt to detect Legionella from various hot springs and the study on detection ability. The 10-μm-pore size MCFS-membrane captured approximately 30% of Legionella spp. in the hot spring. Genes were isolated from most Legionella in the MCFS-membrane when an electric field of 20V/cm was applied to the membrane before PCR amplification. In addition to detecting Legionella, this technique also facilitated bacterial concentration and genetic isolation. Amplification of DNA was not inhibited by hot spring components or proteins and lipids derived from humans as these were removed by the MCFS-membrane. The time required for bacterial identification using this method was shortened to four hours. And this time, the capture rate was 30%. However, it rises until about 100% when the circulation system is used. Thus, this method was suggested that amelioration of the detection ability is easy.
北海道工業大学の論文
2007-03-26

導電性平膜と遺伝子工学的手法を用いたレジオネラ菌検出とその検出感度に関する検討

スポンサーリンク

概要

著者

関連論文

スポンサーリンク