Title:
微生物検出法及び微生物検出キット
Document Type and Number:
Japanese Patent JP4378537
Kind Code:
B2
Abstract:
A live cell of microorganism in a test sample is detected by the following steps of: a) adding a cross-linker capable of cross-linking a DNA by irradiation with light having a wavelength of 350 nm to 700 nm to the test sample; b) irradiating the test sample to which the cross-linker is added with light having a wavelength of 350 nm to 700 nm; c) removing the cross-linker contained in the test sample irradiated with light; d) adding a medium to the test sample from which the cross-linker is removed and incubating the test sample; e) adding again the cross-linker capable of cross-linking a DNA by irradiation with light having a wavelength of 350 nm to 700 nm to the incubated test sample; f) irradiating the test sample to which the cross-linker is added with light having a wavelength of 350 nm to 700 nm; g) extracting a DNA from the test sample and amplifying a target region of the extracted DNA by a nucleic acid amplification method; and h) analyzing the amplified product.
Inventors:
Shinichi Yoshida
Takashi Soejima
Takashi Soejima
Application Number:
JP2008551589A
Publication Date:
December 09, 2009
Filing Date:
August 01, 2008
Export Citation:
Assignee:
Kyushu University
Morinaga Milk Industry Co., Ltd.
Morinaga Milk Industry Co., Ltd.
International Classes:
C12Q1/68; C12N15/09
Other References:
Rudi K. et al.,Use of ethidium monoazide and PCR in combination for quantification of viable and dead cells in comp,Applied and Environmental Microbiology,2005年,Vol.71,p.1018-1024
Rudi K. et al.,Development and application of new nucleic acid-based technologies for microbial community analyses,International Journal of Food Microbiology,2002年,Vol.78,p.171-180
Soejima T. et al.,Photoactivated ethidium monoazide directly cleaves bacterial DNA and is applied to PCR for discrimin,Microbiology and Immunology,2007年 8月,Vol.51,p.763-775
Nogva H. et al.,Ethidium monoazide for DNA-based differentiation of viable and dead bacteria by 5'-nuclease PCR.,BioTechniques,2003年,Vol.34,p.804-813
Rudi K. et al.,Detection of viable and dead Listeria monocytogenes on gouda-like cheeses by real-time PCR.,Letters in Applied Microbiology,2005年,Vol.40,p.301-306
Rudi K. et al.,Development and application of new nucleic acid-based technologies for microbial community analyses,International Journal of Food Microbiology,2002年,Vol.78,p.171-180
Soejima T. et al.,Photoactivated ethidium monoazide directly cleaves bacterial DNA and is applied to PCR for discrimin,Microbiology and Immunology,2007年 8月,Vol.51,p.763-775
Nogva H. et al.,Ethidium monoazide for DNA-based differentiation of viable and dead bacteria by 5'-nuclease PCR.,BioTechniques,2003年,Vol.34,p.804-813
Rudi K. et al.,Detection of viable and dead Listeria monocytogenes on gouda-like cheeses by real-time PCR.,Letters in Applied Microbiology,2005年,Vol.40,p.301-306
Attorney, Agent or Firm:
Yoshiyuki Kawaguchi
Hidemi Matsukura
Tsutomu Toyama
Shinichi Sanuki
Takeshi Niwa
Tomoko Tsujida
Hidemi Matsukura
Tsutomu Toyama
Shinichi Sanuki
Takeshi Niwa
Tomoko Tsujida