Pretreatment method and nucleic-acid-extracting kit usable for the same
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/68
C12Q-001/6806
C12N-015/09
C12N-015/10
출원번호
15505329
(2015-09-02)
등록번호
10316350
(2019-06-11)
우선권정보
JP-2014-179111 (2014-09-03)
국제출원번호
PCT/JP2015/074927
(2015-09-02)
국제공개번호
WO2016/035812
(2016-03-10)
발명자
/ 주소
Yamakawa, Kazutomi
Nagano, Takashi
출원인 / 주소
MIZUHO MEDY CO., LTD.
대리인 / 주소
Wenderoth, Lind & Ponack, L.L.P.
인용정보
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초록▼
The present invention provides a method capable of realizing pretreatment process of genetic screening according to a POCT mode. The method includes: making a sample, extraction liquid for extracting nucleic-acid contained in the sample, silica particles, and a filtering material contact with each o
The present invention provides a method capable of realizing pretreatment process of genetic screening according to a POCT mode. The method includes: making a sample, extraction liquid for extracting nucleic-acid contained in the sample, silica particles, and a filtering material contact with each other; making the filtering material carry composite material of the nucleic-acid and the silica particles thereon; and then delivering the filtering material to a nucleic-acid-amplifying process by means of reaction solution for amplifying nucleic-acid, wherein particle diameters of the silica particles and concentration of the silica particles in the reaction solution for amplifying nucleic-acid are set up within a predetermined range.
대표청구항▼
1. A pretreatment method of extracting nucleic-acid by means of a solid-phase extraction method prior to a nucleic-acid-amplifying process, the pretreatment method comprising: making a sample, extraction liquid for extracting nucleic-acid contained in the sample, silica particles, and a filtering ma
1. A pretreatment method of extracting nucleic-acid by means of a solid-phase extraction method prior to a nucleic-acid-amplifying process, the pretreatment method comprising: making a sample, extraction liquid for extracting nucleic-acid contained in the sample, silica particles, and a filtering material contact with each other;making the filtering material carry composite material of the nucleic-acid and the silica particles thereon; andthen delivering the filtering material to a nucleic-acid-amplifying process, the nucleic-acid-amplifying process using a reaction solution for amplifying nucleic-acid,wherein particle diameters of the silica particles and concentration of the silica particles in the reaction solution for amplifying nucleic-acid are set up within a predetermined range;wherein the predetermined range allows, in advance of the nucleic-acid-amplifying process, omission of both a drying process and an elution process;wherein the predetermined range is defined as follows:5% through 38% of the nucleic-acid is recovered from the sample when based on 100% of nucleic-acid contained in the sample;the concentration of the silica particles is 0.0625 μg/μl through 4 μg/μl;an average particle diameter of the silica particles is 0.01 μm through 100 μm; anda surface area calculated based on the average particle diameter is 1×104 μm2 through 1×108 μm2; andthe pretreatment method further comprises:a first extraction process of adding the sample to the extraction liquid to extract the nucleic-acid contained in the sample;a first absorption process of: making the silica particles contact with the extracted nucleic-acid to obtain the composite material of the nucleic-acid and the silica particles; and making the composite material contact with the filtering material;a first washing process of: washing the composite material and the filtering material with purified water; and delivering the washed composite material and the washed filtering material under a wet condition to the nucleic-acid-amplifying process; andwherein, prior to the delivering the filtering material to the nucleic-acid-amplifying process, the composite material is collected with the filtering material in a manner such that the composite material is on the filtering material. 2. The pretreatment method as defined in claim 1, wherein, prior to the delivering the filtering material to the nucleic-acid-amplifying process, the composite material is separated from the filtering material, and then the separated composite material is delivered to the nucleic-acid-amplifying process. 3. The pretreatment method as defined in claim 1, wherein the first absorption process and the first washing process are performed at the same time. 4. The pretreatment method as defined in claim 1, wherein the first extraction process and the first absorption process are performed at the same time. 5. The pretreatment method as defined in claim 1, wherein the first extraction process, the first absorption process, and the first washing process are performed at the same time. 6. A pretreatment method of extracting nucleic-acid by means of a solid-phase extraction method prior to a nucleic-acid-amplifying process, the pretreatment method comprising: making a sample, extraction liquid for extracting nucleic-acid contained in the sample, silica particles, and a filtering material contact with each other;making the filtering material carry composite material of the nucleic-acid and the silica particles thereon; andthen delivering the filtering material to a nucleic-acid-amplifying process, the nucleic-acid-amplifying process using a reaction solution for amplifying nucleic-acid,wherein particle diameters of the silica particles and concentration of the silica particles in the reaction solution for amplifying nucleic-acid are set up within a predetermined range;wherein the predetermined range allows, in advance of the nucleic-acid-amplifying process, omission of both a drying process and an elution process;wherein the predetermined range is defined as follows:5% through 38% of the nucleic-acid is recovered from the sample when based on 100% of nucleic-acid contained in the sample:the concentration of the silica particles is 0.0625 μg/μl through 1 μg/μl;an average particle diameter of the silica particles is 0.01 μm through 10 μm; anda surface area calculated based on the average particle diameter is 1×105 μm2 through 5×107 μm2; andthe pretreatment method further comprises:a first extraction process of adding the sample to the extraction liquid to extract the nucleic-acid contained in the sample;a first absorption process of: making the silica particles contact with the extracted nucleic-acid to obtain the composite material of the nucleic-acid and the silica particles; and making the composite material contact with the filtering material;a first washing process of: washing the composite material and the filtering material with purified water; and delivering the washed composite material and the washed filtering material under a wet condition to the nucleic-acid-amplifying process; andwherein, prior to the delivering the filtering material to the nucleic-acid-amplifying process, the composite material is collected with the filtering material in a manner such that the composite material is on the filtering material. 7. The pretreatment method as defined in claim 6, wherein, prior to the delivering the filtering material to the nucleic-acid-amplifying process, the composite material is separated from the filtering material, and then the separated composite material is delivered to the nucleic-acid-amplifying process. 8. The pretreatment method as defined in claim 6, wherein the first absorption process and the first washing process are performed at the same time. 9. The pretreatment method as defined in claim 6, wherein the first extraction process and the first absorption process are performed at the same time. 10. The pretreatment method as defined in claim 6, wherein the first extraction process, the first absorption process, and the first washing process are performed at the same time.
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