JP4290118B2 - 異形成と新生物病巣または前新生物病巣を区別するための方法 - Google Patents
異形成と新生物病巣または前新生物病巣を区別するための方法 Download PDFInfo
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Description
a)HPV遺伝子産物の検出のための試薬
b)p16INK4a過剰発現の検出のための試薬
c)検出反応を成し遂げるために通常使用される試薬および緩衝液(例えば、緩衝液、検出マーカー、キャリア物質など)
d)ポジティブコントロール反応を成し遂げるためのp16INK4aサンプル
e)ポジティブコントロール反応を成し遂げるためのHPV遺伝子産物
HPV遺伝子産物および/またはp16INK4aの検出のための試薬は、HPV遺伝子産物および/またはp16INK4a分子に結合し得る任意の薬剤を含み得る。このような試薬としては、タンパク質、ポリペプチド、核酸、ペプチド核酸、糖タンパク質、プロテオグリカン、多糖類または脂質が挙げられる。
子宮頚部のスミアを、p16INK4aに特異的な抗体およびHPV E2タンパク質に特異的なポリクローナル抗体を使用して、免疫細胞学的に染色した。
子宮頚部のスミアを、p16INK4aおよびHPV E2およびL2のmRNAレベルについて、インサイチュ染色反応で、半定量的に分析し得る。この染色反応を、以下のように実行する:
再水和のために、スプレー固定したスミアを、振動デバイス上にある新鮮な50%EtOH中でインキュベートする。この固定手順により産生したPEGフィルムを、激しい洗浄により除去する。次に、このスミアを水中ですすぐ。このスミアを、プロテイナーゼK(PBS中に10μg/ml)とともに10分間37℃で培養する。次いで、このスライドを、洗浄緩衝液(PBS/0.1% Tween20)に移し、そして最終的に脂質ペンで囲む。
子宮頚部のスミアのThinPrep(登録商標)薄層を、p16INK4aに特異的な抗体およびHPV E7タンパク質に特異的モノクローナル抗体を使用して、免疫細胞学的に染色する。
従って、このスライドを、水浴中で40分間95〜98℃で温め、20分間室温に冷まし、洗浄緩衝液(Dakocytomation、S3006)に移し、そして最終的に脂質ペンで囲む。
Claims (23)
- 細胞学的試験手順の過程で、生物学的サンプルにおけるp16INK4aを過剰発現する異形成とp16INK4aを過剰発現する新生物病巣または前新生物病巣を区別する方法であって、以下、
a.該生物学的サンプルにおける、p16INK4aの過剰発現細胞の存在または非存在を決定する工程;
b.該生物学的サンプルにおける、少なくとも1つのハイリスクなHPV遺伝子産物を発現する細胞の存在または非存在を決定する工程;
c.p16INK4aを過剰発現する細胞とハイリスクのHPV遺伝子産物を発現する細胞が同時に存在すること、またはp16INK4aを過剰発現する細胞が単独で存在することを評価する工程;
d.ここで、該p16INK4aを過剰発現する細胞とハイリスクのHPV遺伝子産物を発現する細胞の同時の存在が、新生物病巣または前新生物病巣を示し、少なくとも1つのHPV遺伝子産物が、HPV E7遺伝子によってコードされるものである、工程
を包含する、方法。 - 請求項1に記載の方法であって、ここで、前記ハイリスクのHPV遺伝子産物が、初期の新生物/前新生物病巣において優位に発現される、方法。
- 請求項1〜2のいずれか1項に記載の方法であって、ここで、前記HPV遺伝子産物が、ポリペプチドまたはRNA分子である、方法。
- 請求項1〜3のいずれか1項に記載の方法であって、ここで、前記新生物病巣または前新生物病巣が、肛門性器道の病巣である、方法。
- 請求項4に記載の方法であって、ここで、前記肛門性器道の病巣が、子宮頚部の病巣である、方法。
- 請求項1〜5のいずれかに記載の方法であって、ここで、前記生物学的サンプルが、肛門性器由来の細胞を含むサンプルである、方法。
- 請求項6に記載の方法であって、ここで、前記細胞が、子宮頚部に由来する細胞である、方法。
- 請求項7に記載の方法であって、ここで、前記生物学的サンプルが、パパニコラウスミアまたは前記子宮頚部の細胞学的調製物である、方法。
- 請求項1〜8のいずれか1項に記載の方法であって、ここで、前記HPV遺伝子産物およびp16INK4a分子の検出は、検出される分子に特異的な少なくとも1つのプローブを使用して実行される、方法。
- 請求項9に記載の方法であって、ここで、前記プローブが、検出可能に標識される、方法。
- 請求項10に記載の方法であって、ここで、前記標識が、ラジオアイソトープ、生体発光化合物、化学発光化合物、蛍光化合物、金属キレート、または酵素からなる群から選択される、方法。
- 請求項9〜11のいずれか1項に記載の方法であって、ここで、前記プローブが、タンパク質および/または核酸である、方法。
- 請求項12に記載の方法であって、ここで、少なくとも1つのプローブが、ハイリスクのHPVコード遺伝子産物またはp16INK4aに対して励惹される抗体である、方法。
- 免疫細胞化学的な染色手順を含む、請求項13に記載の方法。
- 請求項12に記載の方法であって、ここで、少なくとも1つのプローブが、ハイリスクのHPV遺伝子産物と特異的にハイブリダイズする核酸である、方法。
- インサイチュハイブリダイゼーション反応を含む、請求項15に記載の方法。
- 核酸増幅反応を含む、請求項15に記載の方法。
- 請求項17に記載の方法であって、ここで、前記核酸増幅反応が、PCRまたはLCRである、方法。
- 請求項1〜18のいずれかに記載の方法であって、ここで、核酸プローブおよびポリペプチドプローブを使用する検出反応が、同時に実行される、方法。
- 請求項1〜19のいずれか1項に記載の方法であって、ここで、前記ハイリスクのHPV遺伝子産物が、癌関連HPVサブタイプのHPV16、18、31、33、35、39、45、51、52、56および58の遺伝子産物である、方法。
- 請求項1〜20のいずれかに記載の方法であって、ここで、少なくとも1つの単細胞において、少なくとも1つのハイリスクHPV遺伝子産物の発現と同時であるp16INK4aの過剰発現が測定される、方法。
- 請求項1〜21のいずれか1項に記載の方法を実行するためのキットであって、該キットは、診断キットまたは研究キットであって、以下:
a.生物学的サンプルにおける、p16INK4aの過剰発現の存在または非存在を検出するためのプローブ、
b.生物学的サンプルにおける、1つ以上のHPV遺伝子産物の発現の存在または非存在を検出するための1つ以上のプローブであって、少なくとも1つのHPV遺伝子産物が、HPV E7遺伝子によってコードされるものである、プローブ、
を含む、キット。 - 請求項22に記載のキットであって、さらに、
a.ポジティブコントロール反応を実行するためのp16INK4aサンプル
b.ポジティブコントロール反応を実行するためのHPV遺伝子産物の1つ以上のサンプル
を含む、キット。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP20020007954 EP1369694A1 (en) | 2002-04-09 | 2002-04-09 | Method for discrimination of metaplasias from neoplastic or preneoplastic lesions |
PCT/EP2003/050096 WO2004005927A1 (en) | 2002-04-09 | 2003-04-08 | Method for discrimination of metaplasias from neoplastic or preneoplastic lesions |
Related Child Applications (1)
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JP2008243422A Division JP4809408B2 (ja) | 2002-04-09 | 2008-09-22 | 異形成と新生物病巣または前新生物病巣を区別するための方法 |
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JP2005523039A JP2005523039A (ja) | 2005-08-04 |
JP4290118B2 true JP4290118B2 (ja) | 2009-07-01 |
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JP2004518777A Expired - Lifetime JP4290118B2 (ja) | 2002-04-09 | 2003-04-08 | 異形成と新生物病巣または前新生物病巣を区別するための方法 |
JP2008243422A Expired - Lifetime JP4809408B2 (ja) | 2002-04-09 | 2008-09-22 | 異形成と新生物病巣または前新生物病巣を区別するための方法 |
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JP2008243422A Expired - Lifetime JP4809408B2 (ja) | 2002-04-09 | 2008-09-22 | 異形成と新生物病巣または前新生物病巣を区別するための方法 |
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US (6) | US7422859B2 (ja) |
EP (3) | EP1369694A1 (ja) |
JP (2) | JP4290118B2 (ja) |
CN (1) | CN1643378A (ja) |
AT (2) | ATE382863T1 (ja) |
AU (1) | AU2003274105A1 (ja) |
DE (2) | DE60318415T2 (ja) |
ES (2) | ES2297209T3 (ja) |
MX (1) | MXPA04009584A (ja) |
WO (1) | WO2004005927A1 (ja) |
Families Citing this family (17)
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EP1369694A1 (en) | 2002-04-09 | 2003-12-10 | MTM Laboratories AG | Method for discrimination of metaplasias from neoplastic or preneoplastic lesions |
ES2215957T3 (es) | 2002-08-01 | 2004-10-16 | Mtm Laboratories Ag | Metodo para diagnostico basado en una solucion. |
EP1422526A1 (en) * | 2002-10-28 | 2004-05-26 | MTM Laboratories AG | Method for improved diagnosis of dysplasias |
US7972776B2 (en) * | 2005-11-15 | 2011-07-05 | Oncohealth Corporation | Protein chips for HPV detection |
US7732166B2 (en) | 2005-11-15 | 2010-06-08 | Oncohealth Corporation | Detection method for human pappilomavirus (HPV) and its application in cervical cancer |
US8968995B2 (en) * | 2008-11-12 | 2015-03-03 | Oncohealth Corp. | Detection, screening, and diagnosis of HPV-associated cancers |
US8278056B2 (en) * | 2008-06-13 | 2012-10-02 | Oncohealth Corp. | Detection of early stages and late stages HPV infection |
US7838215B2 (en) * | 2007-09-25 | 2010-11-23 | Canvir, Inc. | Advanced cervical cell screening methods |
EP2262911B1 (en) * | 2008-04-17 | 2016-10-12 | QIAGEN Gaithersburg, Inc. | Compositions, methods, and kits using synthetic probes for determining the presence of a target nucleic acid |
WO2010025928A1 (en) * | 2008-09-04 | 2010-03-11 | Mtm Laboratories Ag | Method for prediction of the progression risk of tumors |
EP2427763A4 (en) | 2009-05-07 | 2013-08-21 | Oncohealth Corp | IDENTIFICATION OF HIGH GRADE OR CIN2 FOR DETECTION, MONITORING AND DIAGNOSIS, AT EARLY STAGES AND ADVANCED STAGES, OF HUMAN PAPILLOMAVIRUS (HPV) AND HPV ASSOCIATED CANCERS |
CN102822672B (zh) | 2010-01-08 | 2015-12-09 | 安科健康公司 | 用于诊断和筛选与hpv有关的癌症的高通量细胞基hpv免疫测定 |
JP2014528066A (ja) * | 2011-09-09 | 2014-10-23 | アムジエン・インコーポレーテツド | 癌治療での、EGFrに結合する薬剤を確立することにおける、ヒトパピローマウイルスステータスの使用 |
CN103076453B (zh) * | 2012-12-24 | 2015-04-15 | 何以丰 | P16免疫细胞化学试剂盒及其制备用多肽序列 |
CA2946534A1 (en) * | 2014-04-28 | 2015-11-05 | Arizona Board Of Regents On Behalf Of Arizona State University | Novel methods, bioassays, and biomarkers for hpv-related conditions |
WO2016207414A1 (en) * | 2015-06-25 | 2016-12-29 | Universitaet Des Saarlandes | Improved diagnostic method for hpv-induced cancer and precancerous lesion screening and diagnosis |
EP3921441A1 (en) * | 2019-02-08 | 2021-12-15 | Charité - Universitätsmedizin Berlin | A method for determining the severity or grade of human papillomavirus (hpv)-induced dysplasia |
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EP1369694A1 (en) | 2002-04-09 | 2003-12-10 | MTM Laboratories AG | Method for discrimination of metaplasias from neoplastic or preneoplastic lesions |
EP1387173A1 (en) | 2002-07-29 | 2004-02-04 | MTM Laboratories AG | Method for improved diagnosis of cervical lesions based on detection of INK4a gene products |
US20050272038A1 (en) | 2002-07-29 | 2005-12-08 | Ruediger Ridder | Compositions and methods for diagnosis and therapy of cancers |
ES2215957T3 (es) | 2002-08-01 | 2004-10-16 | Mtm Laboratories Ag | Metodo para diagnostico basado en una solucion. |
EP1422526A1 (en) | 2002-10-28 | 2004-05-26 | MTM Laboratories AG | Method for improved diagnosis of dysplasias |
US7361460B2 (en) | 2003-04-11 | 2008-04-22 | Digene Corporation | Approach to molecular diagnosis of human papillomavirus-related diseases |
WO2010025928A1 (en) | 2008-09-04 | 2010-03-11 | Mtm Laboratories Ag | Method for prediction of the progression risk of tumors |
-
2002
- 2002-04-09 EP EP20020007954 patent/EP1369694A1/en not_active Withdrawn
-
2003
- 2003-04-08 AU AU2003274105A patent/AU2003274105A1/en not_active Abandoned
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Also Published As
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MXPA04009584A (es) | 2005-01-11 |
US20090104609A1 (en) | 2009-04-23 |
DE60335922D1 (de) | 2011-03-10 |
DE60318415T2 (de) | 2009-01-08 |
JP4809408B2 (ja) | 2011-11-09 |
EP1821102A1 (en) | 2007-08-22 |
US20120052510A1 (en) | 2012-03-01 |
US20050255468A1 (en) | 2005-11-17 |
EP1493032A1 (en) | 2005-01-05 |
AU2003274105A1 (en) | 2004-01-23 |
US10822660B2 (en) | 2020-11-03 |
ATE382863T1 (de) | 2008-01-15 |
US8043819B2 (en) | 2011-10-25 |
US7422859B2 (en) | 2008-09-09 |
CN1643378A (zh) | 2005-07-20 |
US11155878B2 (en) | 2021-10-26 |
EP1493032B1 (en) | 2008-01-02 |
EP1369694A1 (en) | 2003-12-10 |
JP2005523039A (ja) | 2005-08-04 |
US8815520B2 (en) | 2014-08-26 |
ES2297209T3 (es) | 2008-05-01 |
WO2004005927A1 (en) | 2004-01-15 |
JP2009000127A (ja) | 2009-01-08 |
US20190177804A1 (en) | 2019-06-13 |
US10246751B2 (en) | 2019-04-02 |
US20140308660A1 (en) | 2014-10-16 |
US20170268068A1 (en) | 2017-09-21 |
ES2360342T3 (es) | 2011-06-03 |
DE60318415D1 (de) | 2008-02-14 |
EP1821102B1 (en) | 2011-01-26 |
ATE497169T1 (de) | 2011-02-15 |
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