Bacterial identification method using RNA of sample bacteria, and kit therefor

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 whic...

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Bibliographische Detailangaben
Hauptverfasser: YANAI Hisaaki, AMANO Koh, MORISHIGE Takashi, ENDO Ayako, TSUJI Kentaro
Format: Patent
Sprache:eng
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Beschreibung
Zusammenfassung: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.