METHOD OF DETECTING TARGET NUCLEIC ACID AND NUCLEIC ACID-PROBE-CARRIER COMPLEX
To provide a technique for separating and detecting a target nucleic acid, which is advantageous for detecting pathogens of infectious diseases, in addition to reliably capturing the target nucleic acid and efficiently removing contaminants.SOLUTION: A method of detecting a target nucleic acid compr...
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Zusammenfassung: | To provide a technique for separating and detecting a target nucleic acid, which is advantageous for detecting pathogens of infectious diseases, in addition to reliably capturing the target nucleic acid and efficiently removing contaminants.SOLUTION: A method of detecting a target nucleic acid comprises: (a) a process of capturing a target nucleic acid by forming a complex in the test sample, which may contain the target nucleic acid, the complex comprising a solid-phase carrier, a probe comprising a capture sequence specifically hybridizing to the target nucleic acid immobilized on the solid-phase carrier and a photoreactive group capable of forming a covalent bond with a base in the target nucleic acid upon light irradiation, and the target nucleic acid bound to the probe by hybridization of the probe to the capture sequence and covalent binding based on the photoreactive group; (b) a separation process of separating the complex from the test sample using the covalent bond and the solid-phase carrier; and (c) a nucleic acid amplification process of carrying out a nucleic acid amplification reaction using the target nucleic acid on the solid-phase carrier as a template.SELECTED DRAWING: Figure 1
【課題】標的核酸の確実な捕捉と夾雑物の効率的な除去とともに、感染症病原体の検出に有利な、標的核酸の分離・検出する技術を提供する。【解決手段】 標的核酸の検出方法は、(a)固相キャリアと、固相キャリアに固定化された、標的核酸に特異的にハイブリダイズする捕捉用配列と光照射により前記標的核酸中の塩基との間で共有結合を形成可能な光反応性基とを備えるプローブと、前記プローブの前記捕捉用配列に対するハイブリダイゼーションと前記光反応性基に基づく共有結合とにより前記プローブに結合した前記標的核酸と、を備える、複合体を、前記標的核酸を含有する可能性のある被験試料中において形成して、前記標的核酸を捕捉する工程、(b)前記被験試料から、前記共有結合と前記固相キャリアとを利用して、前記複合体を分離する分離工程、及び(c)前記固相キャリア上の前記標的核酸を鋳型とする核酸増幅反応を実施する核酸増幅工程、を備える。【選択図】図1 |
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