IMPROVED METHOD FOR VIRUS DETECTION

• Main Author: YOSHIKANE TAKASHI
• Format: Patent
• Language: English; Japanese
• Published: 27.09.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

To provide a method and a kit that enable the detection of the presence or absence of virus by directly mixing a one-enzyme-based one-step RT-PCR reaction solution and a sample to perform one-step RT-PCR, without isolating nucleic acids from the sample or preheating the sample.SOLUTION: As a method for testing RNA viruses in a sample, provided is a method for testing the presence or absence of RNA viruses in a sample, characterized by involving the following steps: (1) mixing a sample that has not undergone nucleic acid isolation treatment or heat treatment with a one-enzyme based one-step RT-PCR reaction solution containing a thermostable DNA polymerase having reverse transcription activity; and (2) performing one-step RT-PCR reaction after sealing a reaction container.SELECTED DRAWING: None 【課題】 試料から核酸の単離や試料の事前の加熱処理を行うことなく、一酵素系１ステップＲＴ－ＰＣＲ反応液と試料とを直接混合し、１ステップＲＴ－ＰＣＲを実施することによりウイルスの有無の検出を可能とする方法およびキットを提供する。【解決手段】 試料中のＲＮＡウイルスの検査方法として、以下の工程：（１）核酸の単離処理も加熱処理も行っていない試料と、逆転写活性を有する耐熱性ＤＮＡポリメラーゼを含む一酵素系１ステップＲＴ－ＰＣＲ反応液とを混合する工程、及び、（２）反応容器を密閉後、１ステップＲＴ－ＰＣＲ反応を実施する工程、を包含することを特徴とする試料中のＲＮＡウイルスの存在の有無を検査するための方法を提供する。【選択図】 なし