JP4863506B2 - イオン交換反応及び微生物閉じ込め手段を利用して試料から微生物を分離する方法、微生物試料の前処理用容器、及び微生物分離装置 - Google Patents
イオン交換反応及び微生物閉じ込め手段を利用して試料から微生物を分離する方法、微生物試料の前処理用容器、及び微生物分離装置 Download PDFInfo
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- C12N1/02—Separating microorganisms from their culture media
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- B01J39/00—Cation exchange; Use of material as cation exchangers; Treatment of material for improving the cation exchange properties
- B01J39/02—Processes using inorganic exchangers
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J39/00—Cation exchange; Use of material as cation exchangers; Treatment of material for improving the cation exchange properties
- B01J39/08—Use of material as cation exchangers; Treatment of material for improving the cation exchange properties
- B01J39/14—Base exchange silicates, e.g. zeolites
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Description
本実施例では、入口と出口とが備えられており、サイズ10mMx23mmであるシリコンチップ上にピラーアレイが形成された表面を有するチャンバを備える流動装置に、細菌細胞を含む試料を流しながら、細胞を前記固体支持体上に付着させ、流れ出る試料中の細菌細胞の数をコロニー計数を通じて決定し、これから前記固体支持体による細菌細胞閉じ込め効率を計算した。前記ピラーアレイにおいて、ピラー間の距離は12μmであり、ピラーの高さが100μmであり、各ピラーの断面は、一辺の長さが25μmである正方形である。
緩衝液:100mMの酢酸ナトリウム緩衝液(pH4.0)。
本実施例では、入口と出口とが備えられており、サイズ10mMx23mmであるシリコンチップ上にピラーアレイが形成された表面を有するチャンバを有する流動装置に、細菌細胞を含む試料を流しながら、細菌細胞を前記固体支持体上に付着させ、流れ出る試料中の細胞の数をコロニー計数を通じて決定し、これから前記固体支持体による細菌細胞閉じ込め効率を計算した。前記ピラーアレイにおいて、ピラー間の距離は12μmであり、ピラーの高さが100μmであり、各ピラーの断面は、一辺の長さが25μmである正方形である。
まず、ゼオライトY(Aldrich社製)(NH4−Y形態)を550℃で2時間焼成してHY(H型ゼオライト)を製造した。
まず、ナトリウム型のゼオライトであるNa−Yとアンモニウム型のゼオライトNH4−Yとを購入した(Aldrich社製)。
まず、ゼオライトY(Aldrich社製)(NH4−Y形態)を550℃で2時間焼成してHY(H型ゼオライト)を製造した。
Claims (19)
- 微生物を含む試料をイオン交換特性を有する無機物質であるH型ゼオライトと接触させ
て、前記試料と前記H型ゼオライトとを反応させる段階と、
前記反応された試料を固体支持体にpH3.0〜6.0の範囲の液体中で接触させる段階
と、を含み、
前記試料は尿であり、
前記固体支持体の表面をオクタデシルジメチル(3−トリメトキシシリルプロピル)ア
ンモニウム、トリデカフルオロテトラヒドロオクチルトリメトキシシラン、またはポリエ
チレンイミントリメトキシシランでコーティングしたことを特徴とする、試料から微生物
を分離する方法。 - 前記固体支持体の表面をオクタデシルジメチル(3−トリメトキシシリルプロピル)ア
ンモニウムでコーティングし、かつポリエチレンイミントリメトキシシランの化合物でコ
ーティングしたことを特徴とする、請求項1に記載の試料から微生物を分離する方法。 - 前記微生物は、細菌、真菌、およびウイルスからなる群から選択される少なくとも一つ
である請求項1または2に記載の試料から微生物を分離する方法。 - 前記固体支持体は、表面に複数個のピラーが形成されているピラー構造を有する固体支
持体、ビーズ形状を有する固体支持体、及び表面に複数個の孔隙が形成されている篩構造
を有する固体支持体からなる群から選択される少なくとも一つであることを特徴とする請
求項1〜3のいずれか1項に記載の試料から微生物を分離する方法。 - 前記ピラーは、アスペクト比として、ピラーの高さ:ピラーの断面の長さが1:1〜2
0:1である請求項4に記載の試料から微生物を分離する方法。 - 前記ピラーは、ピラーの高さと最隣接するピラー間の距離との比率が1:1〜25:1
である請求項4または5に記載の試料から微生物を分離する方法。 - 前記ピラーは、最隣接するピラー間の距離が5μm〜100μmの範囲を有する請求項
4〜6のいずれか1項に記載の試料から微生物を分離する方法。 - 前記固体支持体は、水接触角が70゜〜95゜である疎水性を有することを特徴とする
請求項1〜7のいずれか1項に記載の試料から微生物を分離する方法。 - 前記反応された試料を固体支持体にpH3.0〜6.0の範囲の液体中で接触させる段
階前に、前記反応された試料をリン酸緩衝液または酢酸緩衝液で希釈する段階をさらに含
む請求項1〜8のいずれか1項に記載の試料から微生物を分離する方法であって、
前記固体支持体の表面をオクタデシルジメチル(3−トリメトキシシリルプロピル)ア
ンモニウム、トリデカフルオロテトラヒドロオクチルトリメトキシシラン、またはポリエ
チレンイミントリメトキシシランでコーティングしていることを特徴とする、請求項1〜
8のいずれか1項に記載の試料から微生物を分離する方法。 - 前記希釈は、99:1〜1:4の割合でなる請求項9に記載の試料から微生物を分離す
る方法。 - H型ゼオライトが含まれている微生物を含む試料前処理用の第1容器と、
前記第1容器と流体移動可能に連結されている微生物閉じ込め手段である固体支持体を備
える第2容器と、を有し、
前記試料は尿であり
前記固体支持体の表面をオクタデシルジメチル(3−トリメトキシシリルプロピル)ア
ンモニウム、トリデカフルオロテトラヒドロオクチルトリメトキシシラン、ポリエチレン
イミントリメトキシシランでコーティングしたことを特徴とする、微生物を含む試料から
微生物を分離する装置。 - 前記第1及び第2容器は、チューブ、マイクロチャンネル、及びマイクロチャンバから
なる群から選択される少なくとも一つである、請求項11に記載の微生物を含む試料から
微生物を分離する装置。 - 前記H型ゼオライトは、前記第1容器に固定化されている、請求項11または請求項12に記載の微生物を含む試料から微生物を分離する装置。
- 前記第2容器は、緩衝液溶液保存部をさらに備える、請求項11または請求項12に記
載の微生物を含む試料から微生物を分離する装置。 - 前記固体支持体は、表面に複数個のピラーが形成されているピラー構造を有する固体支
持体、ビーズ形状を有する固体支持体、及び表面に複数個の孔隙が形成されている篩構造
を有する固体支持体からなる群から選択される少なくとも一つである、請求項11に記載
の微生物を含む試料から微生物を分離する装置。 - 前記ピラーは、アスペクト比として、ピラーの高さ:ピラーの断面の長さが1:1〜2
0:1である、請求項15に記載の微生物を含む試料から微生物を分離する装置。 - 前記ピラーは、ピラーの高さと最隣接ピラー間の距離との比率が1:1〜25:1であ
る、請求項15または16に記載の微生物を含む試料から微生物を分離する装置。 - 前記ピラーは、ピラー間の距離が5μm〜100μmの範囲を有する、請求項15〜17のいずれか1項に記載の微生物を含む試料から微生物を分離する装置。
- 前記固体支持体は、水接触角が70゜〜95゜の疎水性を有することを特徴とする、請
求項15〜18のいずれか1項に記載の試料から微生物を分離する装置。
Applications Claiming Priority (10)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR10-2006-0079054 | 2006-08-21 | ||
KR1020060079055A KR100813265B1 (ko) | 2006-08-21 | 2006-08-21 | 비평면 형상의 고체 지지체를 이용하여 미생물로부터핵산을 증폭하는 방법 |
KR10-2006-0079055 | 2006-08-21 | ||
KR1020060079053A KR100917465B1 (ko) | 2006-08-21 | 2006-08-21 | 비평면 형상의 고체 지지체를 이용하여 미생물을 분리하는방법 및 미생물분리 장치 |
KR10-2006-0079053 | 2006-08-21 | ||
KR1020060079056A KR100837401B1 (ko) | 2006-08-21 | 2006-08-21 | 비평면 형상의 고체 지지체를 이용하여 미생물로부터핵산을 분리하는 방법,상기 분리된 핵산을 주형으로 하여핵산을 증폭하는 방법 및 상기 고체 지지체를 포함하는핵산 분리 및 증폭 장치 |
KR1020060079054A KR100813264B1 (ko) | 2006-08-21 | 2006-08-21 | 비평면 형상의 고체 지지체를 이용하여 미생물로부터핵산을 증폭하는 방법 |
KR10-2006-0079056 | 2006-08-21 | ||
KR10-2006-0092919 | 2006-09-25 | ||
KR1020060092919A KR100813268B1 (ko) | 2006-09-25 | 2006-09-25 | 이온교환반응 및 미생물 포획 수단을 이용하여 시료로부터미생물을 분리하는 방법, 미생물 시료의 전처리용 용기 및미생물분리 장치 |
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EP1892288A3 (en) | 2008-04-02 |
EP1892288B1 (en) | 2010-05-26 |
US8557564B2 (en) | 2013-10-15 |
JP2008048737A (ja) | 2008-03-06 |
DE602007006731D1 (de) | 2010-07-08 |
US20080044885A1 (en) | 2008-02-21 |
US20120088286A1 (en) | 2012-04-12 |
US8158411B2 (en) | 2012-04-17 |
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