JP2009268404A - 木片からのマツノザイセンチュウのdna抽出方法、マツノザイセンチュウのlampプライマーセット、および木片からのマツノザイセンチュウの検出方法 - Google Patents
木片からのマツノザイセンチュウのdna抽出方法、マツノザイセンチュウのlampプライマーセット、および木片からのマツノザイセンチュウの検出方法 Download PDFInfo
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Abstract
【解決手段】採取されたマツノザイセンチュウが含まれる木片からのマツノザイセンチュウのDNA抽出方法、マツノザイセンチュウのDNAの特定領域にアニーリングするプライマーからなるLAMPプライマーセット、および当該プライマーセットを用いてLAMP法によるDNAの増幅を行い検出する木片からのマツノザイセンチュウの検出方法。
【選択図】なし
Description
真宮靖治・二井一禎・小坂肇・神崎菜摘(2004)材線虫.(線虫学実験法. 日本線虫学会編、日本線虫学会、 茨城). 134-153.
(実施例1)
2×Reaction Mix : 12.5μl
Primer FIP (配列表1) : 1.0μl (40 pmol/μl)
Primer BIP (配列表2) : 1.0μl (40 pmol/μl)
Primer F3 (配列表3) : 1.0μl (5 pmol/μl)
Primer B3 (配列表4) : 1.0μl (5 pmol/μl)
Primer Loop-F (配列表5) : 1.0μl (20 pmol/μl)
Bst DNA Polymerase : 1.0μl
蛍光・目視検出試薬 : 1.0μl
蒸留水 : 3.5μl
計 : 23.0μl
(実施例2および実施例3)
GoTaq green Master Mix :10.0μl
Bx18S1)(配列表6) :2.5μl(2pmol/μl)
Bx28S1)(配列表7) :2.5μl(2pmol/μl)
蒸留水 :3.0μl
計 :18.0μl
1)Aikawa, T., Kikuchi, T. and Kosaka, H. (2003) Demonstration of interbreeding between virulent and avirulent populations of Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae) by PCR-RFLP method. Appl. Entomol. Zool. 38: 565-569.
Claims (6)
- マツノザイセンチュウが含まれる木片をケラチンを分解する酵素を含む溶液中に浸し、前記マツノザイセンチュウのDNAを抽出することを特徴とする木片からのマツノザイセンチュウのDNA抽出方法。
- マツノザイセンチュウのrDNAの特定領域にアニーリング可能な配列表1から4に示されたそれぞれ塩基配列F3、B3、FIP、およびBIPのプライマーからなることを特徴とするLAMPプライマーセット。
- 配列表5に示された塩基配列Loop−Fのプライマーをさらに含むことを特徴とする請求項2に記載のLAMPプライマーセット。
- マツノザイセンチュウの検出方法であって、
前記マツノザイセンチュウのrDNAの特定領域を請求項2または3に記載のLAMPプライマーセットを用いてLAMP法により増幅して検出する段階を備えることを特徴とするマツノザイセンチュウの検出方法。 - マツノザイセンチュウの検出方法であって、
前記マツノザイセンチュウが含まれる木片をケラチンを分解する酵素を含む溶液中に浸し、前記マツノザイセンチュウのDNAを抽出する段階と、
抽出した前記マツノザイセンチュウのDNAを増幅して検出する段階とを備えることを特徴とするマツノザイセンチュウの検出方法。 - 前記マツノザイセンチュウのDNAを増幅して検出する段階は、
前記マツノザイセンチュウのrDNAの特定領域を請求項2または3に記載のLAMPプライマーセットを用いてLAMP法により増幅して検出することを特徴とする請求項5に記載のマツノザイセンチュウの検出方法。
Priority Applications (8)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2008121316A JP5540277B2 (ja) | 2008-05-07 | 2008-05-07 | 木片からのマツノザイセンチュウのdna抽出方法、マツノザイセンチュウのlampプライマーセット、および木片からのマツノザイセンチュウの検出方法 |
KR1020107027367A KR101572143B1 (ko) | 2008-05-07 | 2008-10-23 | 나무 조각으로부터의 소나무 재선충의 dna 추출 방법, 소나무 재선충의 lamp 프라이머 세트 및 나무 조각으로부터의 소나무 재선충 검출 방법 |
US12/989,870 US8318435B2 (en) | 2008-05-07 | 2008-10-23 | DNA extraction method for bursaphelenchus xylophilus from wood chips, lamp primer set for bursaphelenchus xylophilus, and detection method for bursaphelenchus xylophilus from wood chips |
PT2008069185A PT2009136456W (pt) | 2008-05-07 | 2008-10-23 | Método de extracção de adn para bursaphelenchus xylophilus a partir de aparas de madeira, conjunto de iniciadores lamp para bursaphelenchus xylophilus e método de detecção para bursaphelenchus xylophilus a partir de aparas de madeira |
PCT/JP2008/069185 WO2009136456A1 (ja) | 2008-05-07 | 2008-10-23 | 木片からのマツノザイセンチュウのdna抽出方法、マツノザイセンチュウのlampプライマーセット、および木片からのマツノザイセンチュウの検出方法 |
CA2722148A CA2722148C (en) | 2008-05-07 | 2008-10-23 | Dna extraction method for bursaphelenchus xylophilus from wood chips, lamp primer set for bursaphelenchus xylophilus, and detection method for bursaphelenchus xylophilus from woodchips |
CN2008801290536A CN102016030B (zh) | 2008-05-07 | 2008-10-23 | 从木片中提取松材线虫的dna的方法、松材线虫的lamp引物组和从木片中检出松材线虫的方法 |
TW097143368A TWI425093B (zh) | 2008-05-07 | 2008-11-10 | Method for Extracting Bursaphelenchus xylophilus from Wood Tablets, LAMP Primer of Bursaphelenchus xylophilus and Method for Detecting Bursaphelenchus xylophilus from Wood |
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JP2008121316A JP5540277B2 (ja) | 2008-05-07 | 2008-05-07 | 木片からのマツノザイセンチュウのdna抽出方法、マツノザイセンチュウのlampプライマーセット、および木片からのマツノザイセンチュウの検出方法 |
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US (1) | US8318435B2 (ja) |
JP (1) | JP5540277B2 (ja) |
KR (1) | KR101572143B1 (ja) |
CN (1) | CN102016030B (ja) |
CA (1) | CA2722148C (ja) |
PT (1) | PT2009136456W (ja) |
TW (1) | TWI425093B (ja) |
WO (1) | WO2009136456A1 (ja) |
Cited By (2)
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CN112322751A (zh) * | 2020-11-19 | 2021-02-05 | 南京耐德高科技有限公司 | 松材线虫核酸快速检测荧光比色二合一试剂盒及检测方法 |
KR102310223B1 (ko) * | 2021-07-21 | 2021-10-08 | 주식회사 에이아이더뉴트리진 | 랩온페이퍼칩을 포함하는 구조물에 적용하기 위한 세포용해용 조성물 |
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CN103555840B (zh) * | 2013-11-04 | 2015-09-09 | 中国林业科学研究院森林生态环境与保护研究所 | 检测松材线虫的方法、检测引物及其lamp检测试剂盒 |
CN103911433B (zh) * | 2014-03-06 | 2015-08-19 | 广东省农业科学院动物卫生研究所 | 嗜水气单胞菌的lamp检测引物组及试剂盒 |
CN103924001B (zh) * | 2014-04-30 | 2016-04-13 | 南京农业大学 | 一种用于山茶根结线虫lamp 快速检测的引物组合物及其应用 |
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KR102148153B1 (ko) | 2019-04-25 | 2020-08-26 | 대한민국 | 소나무재선충 감염목 조기 진단 유전자 및 이를 이용한 진단 키트, 진단방법 |
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
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CN112322751A (zh) * | 2020-11-19 | 2021-02-05 | 南京耐德高科技有限公司 | 松材线虫核酸快速检测荧光比色二合一试剂盒及检测方法 |
KR102310223B1 (ko) * | 2021-07-21 | 2021-10-08 | 주식회사 에이아이더뉴트리진 | 랩온페이퍼칩을 포함하는 구조물에 적용하기 위한 세포용해용 조성물 |
WO2023003137A1 (ko) * | 2021-07-21 | 2023-01-26 | 주식회사 에이아이더뉴트리진 | 랩온페이퍼칩을 포함하는 구조물에 적용하기 위한 세포용해용 조성물 |
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CA2722148A1 (en) | 2009-11-12 |
CA2722148C (en) | 2016-02-16 |
TW200946681A (en) | 2009-11-16 |
WO2009136456A1 (ja) | 2009-11-12 |
CN102016030A (zh) | 2011-04-13 |
US20110159503A1 (en) | 2011-06-30 |
CN102016030B (zh) | 2012-08-22 |
TWI425093B (zh) | 2014-02-01 |
KR101572143B1 (ko) | 2015-11-26 |
KR20110003397A (ko) | 2011-01-11 |
PT2009136456W (pt) | 2011-09-07 |
US8318435B2 (en) | 2012-11-27 |
JP5540277B2 (ja) | 2014-07-02 |
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