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WO2003102208A3 - Détection rapide de toxines bt-cry - Google Patents

Détection rapide de toxines bt-cry Download PDF

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Publication number
WO2003102208A3
WO2003102208A3 PCT/IN2003/000199 IN0300199W WO03102208A3 WO 2003102208 A3 WO2003102208 A3 WO 2003102208A3 IN 0300199 W IN0300199 W IN 0300199W WO 03102208 A3 WO03102208 A3 WO 03102208A3
Authority
WO
WIPO (PCT)
Prior art keywords
cry
detection
cry2aa
strips
membrane
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/IN2003/000199
Other languages
English (en)
Other versions
WO2003102208A2 (fr
Inventor
Keshav Raj Kranthi
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Indian Council of Agricultural Research
Original Assignee
Indian Council of Agricultural Research
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Indian Council of Agricultural Research filed Critical Indian Council of Agricultural Research
Priority to AU2003249576A priority Critical patent/AU2003249576A1/en
Priority to MXPA04011769A priority patent/MXPA04011769A/es
Publication of WO2003102208A2 publication Critical patent/WO2003102208A2/fr
Publication of WO2003102208A3 publication Critical patent/WO2003102208A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/32—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Bacillus (G)
    • G01N2333/325—Bacillus thuringiensis crystal protein (delta-endotoxin)

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention est basée sur l'utilisation 1) de deux IgGs polyclonaux contre deux toxines Cry, 2) de récepteurs de toxines Cry isolés à partir d'un lépidoptère à l'état larvé et 3) de méthodes manuelles de marquage afin de produire des bandes immuno-chromatographiques utiles pour détecter une variété d'analytes. Les bandes immuno-chromatographiques qui favorisent la détection rapide des analytes sont coûteuses lorsqu'elles sont produites à l'aide d'anticorps monoclonaux et ne sont pas abordables pour des fermiers de l'Inde et des vulgarisateurs agricoles. Les principaux objectifs de la méthode actuelle sont de simplifier le développement de la méthode de détection immuno-chromatographique permettant de détecter des toxines Cry (Bt) à l'aide d'anticorps polyclonaux purifiés par affinité spécifiques de l'analyte, et de fournir une méthode robuste et facile appropriée permettant de préparer des bandes de détection des toxines Cry1Ac/Cry1Ab/Cry2Aa/Cry2Ab à un coût abordable, dans les conditions de l'Inde. Un anticorps anti-Bt (anti-Cry1 Ac ou anti-Cry2Aa/Ab ou les deux) ou des protéines de récepteur de Cry sont immobilisés sur une membrane de nitrate de cellulose par marquage manuel. La membrane est bloquée à l'aide de caséine ou de BSA et de conservateurs. L'anti-Bt (anti-Cry1 Ac ou anti-Cry2Aa/Ab ou les deux) est marqué au moyen d'or et adsorbé sur une membrane en fibre de verre placée au niveau inférieur de la membrane de façon à se chevaucher sur 2mm. La spécificité et la précision du dosage est grandement améliorée du fait de l'utilisation d'un IgG polyclonal purifié à affinité antigène et à affinité protéique A développé dans deux animaux différents (chèvre et lapin). Le schéma de l'ensemble de bandes d'écoulement latéral de détection de toxine Cry est présenté en annexe. Les bandes ainsi produites représentent des dispositifs rapides et sensibles et des méthodes permettant de détecter la présence de Cry1Ac/Cry1Ab/Cry2Aa/Cry2Ab et de toxines cristalline soit à partir de plantes transgéniques soit à partir de produits fermentés Bt. Ces méthodes et ces dispositifs présentent une sensibilité élevée permettant de détecter simultanément une ou deux toxines Cry sur une bande unique. L'utilisation des ces méthodes fournit un système de dosage impliquant un nombre minimum d'étapes, et produit des résultats fiables même lorsque des personnes non entraînées les utilisent.
PCT/IN2003/000199 2002-05-31 2003-05-29 Détection rapide de toxines bt-cry Ceased WO2003102208A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU2003249576A AU2003249576A1 (en) 2002-05-31 2003-05-29 Rapid detection of bt-cry toxins
MXPA04011769A MXPA04011769A (es) 2002-05-31 2003-05-29 Deteccion rapida de toxinas cry de bacillus thuringiensis.

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IN600/DEL/02 2002-05-31
IN600DE2002 2002-05-31

Publications (2)

Publication Number Publication Date
WO2003102208A2 WO2003102208A2 (fr) 2003-12-11
WO2003102208A3 true WO2003102208A3 (fr) 2004-03-25

Family

ID=29596804

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IN2003/000199 Ceased WO2003102208A2 (fr) 2002-05-31 2003-05-29 Détection rapide de toxines bt-cry

Country Status (6)

Country Link
KR (1) KR20050026396A (fr)
CN (1) CN100403028C (fr)
AU (1) AU2003249576A1 (fr)
MX (1) MXPA04011769A (fr)
WO (1) WO2003102208A2 (fr)
ZA (1) ZA200410268B (fr)

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007000048A1 (fr) * 2005-06-28 2007-01-04 Zbx Corporation Matrice de membranes et dispositif analytique
CN100348616C (zh) * 2005-12-12 2007-11-14 中国农业大学 一种Bt Cry1A抗体及其制备方法与专用抗原及应用
AU2009343906B2 (en) * 2009-04-09 2013-07-04 Hitachi Chemical Diagnostics Systems Co., Ltd. Detector and detection method
CN103197075B (zh) * 2012-01-16 2014-10-08 华中农业大学 利用量子点检测转基因水稻中Bt蛋白的方法
DK3680000T3 (da) 2013-03-15 2024-04-02 Glaxosmithkline Ip Dev Ltd Fremgangsmåder til oprensning af antistoffer
CN105693856A (zh) * 2016-04-25 2016-06-22 江苏省农业科学院 一种单克隆抗体及其分泌该单克隆抗体的细胞株、制备方法与应用
CN109689675A (zh) 2016-09-07 2019-04-26 葛兰素史密斯克莱知识产权发展有限公司 纯化抗体的方法
CN106674334B (zh) * 2017-02-08 2020-09-01 金陵科技学院 一种结合Cry2Ad的环七肽及其编码基因和应用
CN108828230B (zh) * 2018-06-21 2021-04-30 北京市农林科学院 核酸层析快速检测转基因产品的方法
CN110346569A (zh) * 2019-06-28 2019-10-18 安徽恩禾生物技术有限公司 一种胸苷激酶化学发光法检测试剂盒及其制备方法
CN114280312B (zh) * 2020-09-27 2023-09-15 河北特温特生物科技发展有限公司 一种用于免疫荧光层析检测的全血分离膜及其制备方法和应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5141850A (en) * 1990-02-07 1992-08-25 Hygeia Sciences, Inc. Porous strip form assay device method
US5712172A (en) * 1995-05-18 1998-01-27 Wyntek Diagnostics, Inc. One step immunochromatographic device and method of use
US6156573A (en) * 1996-11-20 2000-12-05 Monsanto Company Hybrid Bacillus thuringiensis δ-endotoxins with novel broad-spectrum insecticidal activity
WO2001044779A2 (fr) * 1999-12-14 2001-06-21 Strategic Diagnostics, Inc. Procede de traitement et de test d'echantillons pulverises au moyen de bandelettes reactives immunochromatographiques

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5712171A (en) * 1995-01-20 1998-01-27 Arqule, Inc. Method of generating a plurality of chemical compounds in a spatially arranged array
PT942985E (pt) * 1996-11-20 2004-12-31 Monsanto Technology Llc Delta-endotoxinas de largo espectro

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5141850A (en) * 1990-02-07 1992-08-25 Hygeia Sciences, Inc. Porous strip form assay device method
US5712172A (en) * 1995-05-18 1998-01-27 Wyntek Diagnostics, Inc. One step immunochromatographic device and method of use
US6156573A (en) * 1996-11-20 2000-12-05 Monsanto Company Hybrid Bacillus thuringiensis δ-endotoxins with novel broad-spectrum insecticidal activity
WO2001044779A2 (fr) * 1999-12-14 2001-06-21 Strategic Diagnostics, Inc. Procede de traitement et de test d'echantillons pulverises au moyen de bandelettes reactives immunochromatographiques

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
AGDIA MOLECULAR DIAGNOSTICS, Catalogue, ImmunoStrip test system for Bt-CrylAb/lAc proteins; ImmunoStrip test system for Bt-Cry3A protein from, http://www.agdia.com (Feb2001) in conjunction with http://web.archive.org. *
ENVIROLOGIX INC., Lateral Flow QuickStix Strip Kit, from http://envirologix.com *
Jenkins, J.L. and Dean D.H. (2001) BMC Biochemistry, vol. 2:12. "Binding specificity of Bacillus thuringiensis CrylAa for purified native Bombyx mori aminopeptidase N and cadherin-like receptors". *
Saxena, D. et al. (1999) Nature, vol. 402, p.480. "Insecticidal toxin in root exudates from Btcorn". *
Stave, J.W. (May/June 2002) Journal of AOAC International, vol. 85(3), pp. 780-786. "Protein immunoassay methods for detection of biotech crops: Applications, limitations and practical considerations". *

Also Published As

Publication number Publication date
CN100403028C (zh) 2008-07-16
AU2003249576A8 (en) 2003-12-19
WO2003102208A2 (fr) 2003-12-11
ZA200410268B (en) 2006-07-26
AU2003249576A1 (en) 2003-12-19
CN1672049A (zh) 2005-09-21
MXPA04011769A (es) 2005-07-26
KR20050026396A (ko) 2005-03-15

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