Title:
BACTERIAL IDENTIFICATION METHOD USING RNA OF SAMPLE BACTERIA, AND KIT THEREFOR
Document Type and Number:
WIPO Patent Application WO/2019/189266
Kind Code:
A1
Abstract:
The purpose of the present invention is to provide an identification method by which it is possible to identify many bacteria, rather than a limited number of bacterial species, without changing the identification reagent and conditions. Another purpose is to provide is an identification method which makes it possible to perform detection and identification with high sensitivity with only one PCR, and to reduce complicated operations and the burden on the operator. The present invention uses a method having the following steps to identify bacteria: (1) a step for using a first reaction system including a reverse transcription primer for preparing cDNA including a base sequence for identifying bacteria to be identified, RNA extracted from bacteria in a sample, and an enzyme having an RNA-dependent DNA polymerase activity, performing a reverse transcription reaction, and obtaining a reaction mixture including the synthesized cDNA and the reverse transcription primer; (2) a step for using a second reaction system including the reaction mixture, a primer pair for synthesizing double-stranded DNA including the base sequence for identifying the bacteria to be identified, and an enzyme having a DNA-dependent DNA polymerase activity, and performing PCR, with the caveat that in the second reaction system, the concentration of the reverse transcription primer supplied from the reaction mixture is 0.08 nM to 20 nM; and (3) a step for detecting the production of double-stranded DNA including the base sequence for identifying the bacteria to be identified from the second reaction system after PCR.
Inventors:
MORISHIGE TAKASHI (JP)
AMANO KOH (JP)
YANAI HISAAKI (JP)
ENDO AYAKO (JP)
TSUJI KENTARO (JP)
AMANO KOH (JP)
YANAI HISAAKI (JP)
ENDO AYAKO (JP)
TSUJI KENTARO (JP)
Application Number:
PCT/JP2019/013021
Publication Date:
October 03, 2019
Filing Date:
March 26, 2019
Export Citation:
Assignee:
MITSUI CHEMICALS INC (JP)
International Classes:
C12Q1/689; C12N15/11
Domestic Patent References:
| WO2015053293A1 | 2015-04-16 | |||
| WO2010082640A1 | 2010-07-22 | |||
| WO2007097323A1 | 2007-08-30 | |||
| WO2010054819A1 | 2010-05-20 | |||
| WO2015053293A1 | 2015-04-16 |
Foreign References:
| JP2010162042A | 2010-07-29 | |||
| JP2018011590A | 2018-01-25 | |||
| JP2012034705A | 2012-02-23 | |||
| JP2012034705A | 2012-02-23 |
Other References:
DRESES-WERRINGLOER, U. ET AL.: "Presistence of Chlamydia trachomatis is induced by ciprofloxacin and olfloxacin in vitro", ANTIMICROB. AGENTS CHEMOTHER., vol. 44, no. 12, 1 December 2000 (2000-12-01), pages 3288 - 3297, XP055639199
S. CHAKRAVORTY ET AL., JOURNAL OF MICROBIOLOGICAL METHODS, vol. 69, 2007, pages 330 - 339
See also references of EP 3778925A4
S. CHAKRAVORTY ET AL., JOURNAL OF MICROBIOLOGICAL METHODS, vol. 69, 2007, pages 330 - 339
See also references of EP 3778925A4
Attorney, Agent or Firm:
MIYAZAKI Teruo et al. (JP)
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