KR900016463A - 완전혈액 또는 pbmc 부분으로부터 핵산을 추출하여 증폭시키며 검사하는 방법 - Google Patents

완전혈액 또는 pbmc 부분으로부터 핵산을 추출하여 증폭시키며 검사하는 방법 Download PDF

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KR900016463A
KR900016463A KR1019900005347A KR900005347A KR900016463A KR 900016463 A KR900016463 A KR 900016463A KR 1019900005347 A KR1019900005347 A KR 1019900005347A KR 900005347 A KR900005347 A KR 900005347A KR 900016463 A KR900016463 A KR 900016463A
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pbmc
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아써 버딕 브센트
데이비스 에케즈 토비어스
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제이. 제프리 헐리
이스트만 코닥 컴퍼니
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Abstract

내용 없음.

Description

완전혈액 또는 PBMC 부분으로부터 핵산을 추출하여 증폭시키며 검사하는 방법
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음

Claims (10)

  1. A. 병원체나 사람의 게놈으로 부터의 핵산을 함유하고 있는 PBMC의 수성 샘플을 물의 비등점 또는 비등점 근처에서 2-15분간 가열하여 샘플내 세포를 용해시키고 이 세포로 부터 핵산을 방출시키며, B. 가열된 샘플로 부터 핵산을 회수하는 단계로 구성되는 것으로 특징되는 PBMC의 수성 샘플로 부터 핵산을 빠르게 추출하는 방법.
  2. A. PBMC의 수성 샘플내 병원체나 사람의 게놈으로 부터 핵산을 추출하고, B. 추출된 핵산을 중합요소 연결반응을 이용해 증폭시키는 것으로 구성되고, 추출과정이 1)PBMC의 수성 샘플을 물의 비등점, 또는 비등점 근처에서 2-15분간 가열하여 샘플내 세포를 용해시키고 이 세포로 부터 핵산을 방줄시키며, 2) 가열된 샘플로부터 핵산을 회수하는 단계로 구성되는 것으로 특징되는, 중합효소 연결반응을 이용해 미리 알고있는 핵산을 증폭시키는 방법.
  3. A. 1)병원체 또는 사람의 게놈의 핵산을 함유하고 있는 PBMC의 수성 샘플을 물의 비등점 또는 비등점 근처에서 2-15분간 가열하여 샘플내 세포를 용해시키고 이 세포로 부터 핵산을 방출시키며 핵산을 변성시키고, 2) 변성된 핵산을 가열된 샘플로 부터 회수하는 단계로 구성된 추출과정, B. 회수된 변성 핵산을 핵산의 분리된 사슬에 상보적인 제1및 제2프라이머(primer)와 접촉시켜 프라이머와 상보적인 사슬과의 혼성화 생성물을 형성시키며, C. 상보체와 분리될때 프라이머의 증대 생성물 합성에서 주형으로 작용할 수 있는, 혼성화 생성물내 프라이머의 제1및 제2증대 생성물을 형성시키고, D. 프라이머 증대 생성물을 합성시켰던 주형으로 부터 프라이머 증대 생성물을 분리하며, E. 분리된 증대 생성물과 미리 알고있는 핵산을 부가적인 제1및 제2프라이머와 접촉시키므로써 핵산을 증폭시켜 상보적인 생성물을 형성시키고, F. 프라이머 증대 생성물을 단계 E에서 형성된 상보적인 생성물로 부터 분리시키며, G. 수성 샘플내 핵산의 존재 표시로서, 증폭된 핵산을 검사하는 것으로 구성된, 두개의 상보적인 사슬을 갖는 핵산의 검사방법.
  4. 제1항 내지 제3항의 어느 한 항에 있어서, HIV- I DNA를 추출, 증폭 또는 검사하는 방법.
  5. 제1항 내지 제4항의 어느 한 항에 있어서, 핵산을 회수하는 것이 가열한 추에 샘플을 원심분리시킴에 의해 성취되는 방법.
  6. A. 완전혈액 샘플을 하나 이상의 다당류로 구성된 염용액파 혼합하고, B. 단계 A의 혼합물을 최소한 물의 비등점 또는 비등점 근천의 온도에서 2-15분간 가열하여 혼합물내 세포를 용해시키고 세포로 부터 핵산을 방출시키며, C. 핵산을 가열된 혼합물로 부터 회수하는 단계로 구성되는, 완전혈액 샘플로 부터 핵산을 게 추출하는 방법.
  7. 제6항에 있어서, 염 용액이 비이온성 계면활성재로 더 구성되는 방법.
  8. 제5항 또는 제7항에 있어서, 사람의 β-글로빈 또는 사람의 백혈구 항원 DNA를 추출하는 방법.
  9. 제6항 내지 제8항의 어느 한 항에 있어서, 핵산이 건조된 완전혈액 샘플로 부터 추출되는 방법.
  10. 제1항 내지 제7항의 어느 한 항에 있어서 가열하는 것이 95°-120℃온도에서 4-12분간 행해지는 방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019900005347A 1989-04-17 1990-04-17 완전혈액 또는 pbmc 부분으로부터 핵산을 추출하여 증폭시키며 검사하는 방법 KR900016463A (ko)

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US33943789A 1989-04-17 1989-04-17
US07/406,222 US5334499A (en) 1989-04-17 1989-09-12 Methods of extracting, amplifying and detecting a nucleic acid from whole blood or PBMC fraction
US406.222 1989-09-12
US339,437 1994-11-14

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KR900016463A true KR900016463A (ko) 1990-11-13

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US (1) US5334499A (ko)
EP (2) EP0620282B1 (ko)
JP (1) JPH072120B2 (ko)
KR (1) KR900016463A (ko)
AT (2) ATE117377T1 (ko)
CA (1) CA2013316A1 (ko)
DE (2) DE69016079T2 (ko)
DK (1) DK0620282T3 (ko)
FI (1) FI901917A0 (ko)

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DE69016079T2 (de) 1995-09-14
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