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WO1999053322A1 - Essai en phase solide pour detecter l'endotoxine - Google Patents

Essai en phase solide pour detecter l'endotoxine Download PDF

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Publication number
WO1999053322A1
WO1999053322A1 PCT/GB1999/001099 GB9901099W WO9953322A1 WO 1999053322 A1 WO1999053322 A1 WO 1999053322A1 GB 9901099 W GB9901099 W GB 9901099W WO 9953322 A1 WO9953322 A1 WO 9953322A1
Authority
WO
WIPO (PCT)
Prior art keywords
endotoxin
solid phase
phase apparatus
indicator
buffer components
Prior art date
Application number
PCT/GB1999/001099
Other languages
English (en)
Inventor
Arezki Mahiout
Original Assignee
Allied Therapeutics Limited
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 Allied Therapeutics Limited filed Critical Allied Therapeutics Limited
Priority to AU34328/99A priority Critical patent/AU3432899A/en
Priority to CA002327169A priority patent/CA2327169A1/fr
Priority to EP99915905A priority patent/EP1042672A1/fr
Publication of WO1999053322A1 publication Critical patent/WO1999053322A1/fr

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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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56911—Bacteria
    • 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
    • G01N33/579—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving limulus lysate
    • 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
    • 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)

Definitions

  • This invention relates to a solid phase test for endotoxin, in ⁇ particular a test such as in a flat sheet or in capillary form for the detection of endotoxin in aqueous solutions. More particularly the invention relates to a direct solid phase chromogenic assay, which permits a quick biochemical identification of endotoxin and is sensitive to picogram endotoxin amounts.
  • Horseshoe crab (limulus) amoebocyte lysate provokes together with endotoxin from gram-negative bacteria a gelatination induced by a stepwise activation of several coagulation factors contained in the lysate.
  • An assay comprising the extract (lysate) of horseshoe crab blood cell (amoebocyte) to induce this gelatination in a reagent glass as a liquid phase system is well known as a method and it is called the limulus test.
  • the application of the LAL to measure endotoxin involves the gelatination of the sample (gelation method, turbidimetry assay) in the reagent glass or a more sensitive method based on a chromogenic substrate and measurement by optical absorption.
  • gelation method turbidimetry assay
  • -Today different LAL from horseshoe crab 2 are - 2 - commercialized through the world: Limulus Polyphemus, Tchypleus tridendatus, Tachypleus glgas and Carcinoscorpius rotundicauda.
  • All commercialized LAL testing methods based on a chromogenic substrate are liquid-phase assays.
  • the principle is that in a reagent glass containing a lysate and sample, bacterial endotoxin contained in the sample initiates activation of a cascade of serine protease (Factor B, Factor C, Proclotting enzyme, Coagulin) enzymes in LAL that cleave the end product coagulogen into a peptide coagulin which produces clotting.
  • a cascade of serine protease Factor C
  • Proclotting enzyme Proclotting enzyme
  • a synthetic peptide having an amino acid sequence in common with the hydrolysis sites of coagulogen, namely the chromogenic substrate Boc-Leu-Gly-Arg-p-nitroanilide (pNA) or the fluoro-genic substrate Boc-Leu-Gly-Arg-4-methylcoumaryl-7-amide.
  • the colourless substrate is rapidly cleaved to form the chromophore p-nitroaniline or the fluorogenic 4-methylcoumaryl-7-amide.
  • Tanaka disclosed a quantitative kit for endotoxin determination which comprises an insoluble carrier on which is immobilized an endotoxin-sensitive factor derived from a limulus amoebocyte which specifically reacts with endotoxin without reacting with ⁇ -glucan.
  • the test system involves the indication of endotoxin when colouring occurs in the liquid phase.
  • the present invention provides an analytical test strip for the detection of endotoxin in aqueous solutions in which the strip is wetted at the bottom with the sample. After a certain period of time, until the sample flows from the bottom to the top, an indicator is added at the top, to develop a red colour which is indicative for the presence or the absence of endotoxin in the sample.
  • the present invention involves an improvement to a quick test strip for the detection of endotoxin.
  • the test strip provides a dry analytical element for endotoxin which comprises an absorbent carrier divided in four portions impregnated with: 1 ) a reagent system of limulus factors; 2) synthetic chromogenic substrate derivatized with p-nitroaniline; 3) a buffer system containing chloride acid and sodium nitrite; 4) a buffer system containing chloride acid and ammonium.
  • the test also includes a solution containing N1 -naphtylethylenediamine which is added at the top of the test trip to develop a coloration by coupling the derivatized p-NA to N1 - napthylethylenediamine as positive results.
  • the present invention provides apparatus and method as defined in the claims, in particular an analysis method comprising applying a aqueous sample solution to the bottom of the described dry analytical element to react with endotoxin, and determining coloration on the top of the dry element by coupling N1 - napthylethyienediamine.
  • the present invention also provides an endotoxin kit using the above described dry analytical element.
  • a further object of this invention is to provide a method for preparing the above mentioned dry analytical element and/or the portions of the dry analytical element for preparing the above described endotoxin test strip. - 4 -
  • the method involved in the invention is a direct solid phase chromogenic calorimetricai detection.
  • Four supports are arranged successively in the same alignment to achieve a flow communication with each other.
  • the supports are mounted in a closed and pyrogen free casting module.
  • the test strip comprises a solid support having four portions (at least three portions) being in flow communication with each other whereby reaction products can flow from one portion to the next.
  • the solid support is preferably shaped in the form of strip, with the first, second, third and fourth portions being arranged on the strip in the same plane.
  • the sizes of the portion may be the same or different.
  • the portions are manufactured to absorb the different chemicals and dried, and attached on the strip in a way that flow communication occurs from one to the next.
  • test strip would be submerged with the sample at the bottom to wet the first portion and allow liquid to flow from bottom to the top of the test strip.
  • sample solution would start to flow by capillary action from one portion to the next higher portion.
  • reactants occurring in each portion would be transported by capillary flow to the last portion to develop a red coloration as a positive result.
  • the first portion contains a solid support and a limulus " amoebocyte lysate or substances containing endotoxin sensitive factors which react with endotoxin to produce coagulin.
  • the method of the immobilization of the lysate or of the endotoxin sensitive factor to the solid support are: 1 ) absorption; 2) adsorption by ionic forces or/and hydrophobic interaction, 3) covalent binding.
  • the procedures for binding the lysate in the solid support are generally known the art.
  • the lysate may be the commercial available limulus amoebocyte lysate such as Limulus Polyphemus, Tchypleus tridendatus, Tachypleus gigas and Carcinoscorpius rotundicauda.
  • a lysate without Factor G may be used in order to avoid a positive false reaction with (1 -3)- ⁇ -D-Glucan contained in the sample or in the solid support.
  • a purified form of a mixture containing factors C, B and proclotting enzymes, which react with- endotoxin to produce coagulin may also be used.
  • a lysate preparation known in the art without Factor G which does not react with (1 -3)_- ⁇ -D-Glucan may also be used.
  • endotoxin activates the bound lysate in the following reaction steps:
  • the clotting enzyme produced through the cascade reaction by the action of activated Factor C or proclotting enzymes is capable of hydrolysing coagulin or an amide linkage of a synthetic peptide substrate at the specific sites intermediate between Arg and Gly or intermediates between Arg and Thr.
  • the solid support which is employed in this test strip is one which is capable of absorbing endotoxin from the sample.
  • the sample containing endotoxin reacts with reagents in the first portion and the reaction products are transported by flow to the dry region of the first support and thus to the second portion of the test strip.
  • the solid support is one which is capable of absorbing both sample and binder.
  • an hydrophillic polymers solid support examples are chromatographic papers, nitrocellulose, dextran and glass fibres.
  • chromatographic hydrophobic polymers are used though if using chromatography papers, it is important to use reagents which do not react with ⁇ -glucan, as described for example in US patent 5,550,030.
  • the second portion contains a solid support and a synthetic substrate.
  • a typical substrate to be immobilized in the second portion is t-butoxycarbonyl-leucyl-glycyl- - 6 - arginine-paranitroanilide (Boc-Leu-Gly-Arg-pNA) to release paranitroanilide.
  • Usable synthetic peptides with a known detectable group are those described in EP-A- 0000063, EP-A-00181 12, WO79/00602, WO82/02382 and US 4, 188,264.
  • synthetic peptide substrates for activated Factor C as well as synthetic peptide substrates for activated clotting enzymes in which a carboxyl group of the C-terminated arginine " is substituted with the colour-developing residue p- nitroaniline, p-(N,N-diethylamine)aniline, p-(N-ethyl-N- ⁇ -hydroxyethyl) aniline can also be used.
  • Possible methods of immobilization of the chromogenic substrate to the solid support are: 1 ) absorption; 2) adsorption by ionic forces or/and hydrophobic interaction, and 3) covalent binding.
  • the support is absorbed with the substrate.
  • the solid support is typically made with the same material as the first support and it is one which is capable of absorbing the liquid and reactants flowing from the first support, and which when wetted is this way, provides for flow and analyte by capillary attraction from the first portion, and through the second portion into the third portion.
  • solution should react with immobilized chromogenic substrate.
  • suitable supports are chromatographic papers, nitrocellulose, dextran and glass fibres. Preferably, chromatographic papers are used.
  • the solid support absorbed with the chromogenic substrate is one which is capable otabsorbing reactants flowing from the first support.
  • the so-called reactants are capable of reacting with the second portion and the reactants with the new reaction products being transported to the dry regions and thus to the third portion of the test strip.
  • the solid support is so manufactured (with attention to the amount of absorbed synthetic chromogenic substrate, size of the portion, material of the solid support) that activated clotting enzyme and/or factor B flowing from the lower portion reacts with substrate in the second portion and the chromophore p-nitroaniline is cleaved. The cleaved p-nitroaniline is then transported to the next portion.
  • the third portion contains a solid support and a buffer containing 1 N HCI and 0.1 % (w/v) sodium nitrite.
  • a buffer containing 1 N HCI and 0.1 % (w/v) sodium nitrite.
  • the support it is impregnated with the buffer system, dried and is made with the same material as the second support.
  • the solid support When dried, the solid support is also one which is capable of absorbing the liquid and the chromophore p-nitroaniline flowing from the second support, and which when wetted is this way, provides for flow from the second portion, and through the third portion into the fourth portion.
  • the fourth portion contains a solid support and a buffer containing ⁇ N HCI containing 0.5% ammonium.
  • a solid support Preferably, to make the support, it is impregnated with the buffer system, and dried.
  • the solid support When absorbed with the buffer system and dried, the solid support is also one which is capable of absorbing the liquid and the chromophore p-nitroaniline flowing from the third support, and which when wetted is this way, provides for flow from the third portion, and through the dried region.
  • the third and fourth portions are those through which when the flowing free p- nitroaniline released by the action of enzymes is derivatized to its diazanium salt, typically formed in the fourth portion.
  • the fifth portion is a neutral portion in which a drop of 0.05% N 1 - naphtylethylenediamine in 40-50% (v/v) is added to form a highly visible red colour by coupling of the derivatized p-NA to N1 -napthylethylenediamine.
  • test strip described in this invention can suitably be prepared from any matrix material through which an aqueous solution can flow by capillarity.
  • Suitable materials for chromatographic strips are paper, nitrocellulose and hydrophillic polymers. Because of the presence of ⁇ -glucan in cellulosic material, paper is not usually suitable without special treatment as it induces false positive results. In this invention, it is desirable to use hydrophillic polymers which allow vertical capillary flow excluding a bibulous lateral flow such as polyethylene sheet material.
  • Fig. 1 shows a schematic representation of a capillary test strip
  • Fig. 2 shows a schematic representation of a paper test strip
  • Fig. 3 shows mode of operation of a test strip
  • Fig. 4 shows variation of substrate sensitivity and resulting lysate activity
  • Fig. 5 shows a quantitative test strip.
  • Example 1 capillary test strip
  • a glass howl capillary glass with a flat side, an internal dimension of 0.25 cm and an external dimension of 0.28 cm and a length of 4 cm has been used as the casting module.
  • a cap which is used as the sample reservoir, whereas a second cap closes the top of the capillary and permits entry of chemicals for the colour reaction.
  • the module is constructed in a way that it remains pyrogen free.
  • the howl module is successively filled with dried hydrophillic porous spherical material (200-500 ⁇ m).
  • the hydrophillic porous polymer is in separate portions, impregnated as follows:
  • Portion 1 hydrophillic polymeric powder was incubated under pyrogen free conditions in 0.5 lU/ml (1 g/ml) of limulus amoebocyte lysate (Charles River- Sulzfel Germany) until saturated absorption of the liquid was reached. After drying, the module has been filled to reach a height of 1 cm. 0.1 cm has then been filled with neutral polymeric powder.
  • Portion 2 10 ⁇ mol of chromogenic substrate (t-butoxycarbonyl-leucyl-glycyl- arginine-paranitroanilide) for endoxin (Pefachrome LAL Code .NR 1 1 179-01 ISO 9001 -1 N29001 Pentapharm AG BASEL CH) was diluted in 6.6 ml pyrogen free water. 1 g of hydrophillic polymeric material was also incubated with 1 ml until saturated absorption was reached. After drying the module has been filled to reach a height of 0.5 cm. 0.1 cm has then been filled with neutral polymeric powder.
  • chromogenic substrate t-butoxycarbonyl-leucyl-glycyl- arginine-paranitroanilide
  • endoxin Pefachrome LAL Code .NR 1 1 179-01 ISO 9001 -1 N29001 Pentapharm AG BASEL CH
  • Tris-HCI is mixed with polymeric powder to obtain a concentration of 0.025 mmol/l and inserted between portions 2 and 3 with a height of 0.5 cm. 0.1 cm has been filled with neutral polymeric powder.
  • Portion 3 Polymeric powder was submerged in a solution of 1 N HCI containing 0.1 % (w/v) sodium nitrite, and dried. A quantity sufficient for 0.5 cm has been inserted into the module. 0.1 cm has been filled with neutral polymeric powder.
  • Portion 4 1 cm of polymeric powder was submerged with in solution of 1 N HCI containing 0.5 % (W/v) ammonium sulfate, dried and has been filled into the module. 0.1 cm has been filled with neutral polymeric powder.
  • Portion 5 2.5 mg of N 1 -naphthlethylenediamine in mixed with 0.5 cm polymeric material.
  • the strip can prepared from any matrix material through which the test fluid can vertically flow by capillarity; as illustrated in figure 2, a polystyrene dipstick was constructed to be fitted in a sterile bottle.
  • the active surface of the test strip was 0.5 cm x 4.2 cm, separated every 0.5 cm between the first portion and the third portion with 0.5 cm x 0.2cm dry Sephadex.
  • the active surface between the sephadex strip was coated with Scotch adhesive transfer tape 969 (3M, USA, 55144)
  • Portion 1 consisted of 0.5cm x 0.5 cm ultra high molecular weight polyethylene sheet material (Porex Technologies) which was incubated for 10 min under pyrogen free conditions in 0.5 lU/ml (1 g/ml) of limulus amoebocyte lysate (Charles River- Sulzfel Germany) . After drying, it was applied to the bottom of the test strip. A neutral 0.5 cm x 0.1 cm portion was added above the neutral sephadex.
  • Portion 2 consisted of 0.5cm x 0.5 cm ultra high molecular weight polyethylene - 10 - sheet material (Porex Technologies) which was spotted with 1 .5 /mol/ml of chromogenic substrate (t-butoxycarbonyl-leucyl-glycyl-arginine-pa-ranitroanilide) for ⁇ ndoxin (Pefachrome LAL Code NR 1 1 1 79-01 ISO 9001 -1 N29001 Pentapharm AG BASEL CH) . The strip was applied just above the neutral sephadex portion.
  • Portion 3 consisted of a 0.5 cm x 0.5 cm strip of filter paper (Schleicher & Shuel 23 SL) which was submerged with solution of 1 N HCI containing 0.1 % (w/v) sodium nitrite, dried and applied above the neutral sephadex portion.
  • Portion 4 consisted of a 0.5 cmx 0.5 cm strip of filter paper (Schleicher & Shuel 23 SL) which was submerged with in solution of 1 N HCI containing 0.5 % (W/v) ammonium sulfate, dried and applied above the third portion.
  • Portion 5 consisted of a 0.5 cmx 0.5 cm strip of filter paper (Schleicher & Shuel 23 SL) applied above the fourth portion.
  • sample 1 add the sample to the first portion of the test strip or immerse the first portion in the sample (example 2) or fill the reservoir with the sample (example 1 ) .
  • clotting enzyme is produced through the cascade reaction by the action of activated Factor C or proclotting enzymes.
  • portion 3 after the .capillary flow through the chromogenic substrate and in the presence of Tris-HCI, the activated clotting enzyme and/or factor B flowing from the lower portion cleaves p-nitroaniline. The cleaved p-nitroaniline being transported to the third and fourth portion and derivatized to its diazonium salt.
  • the diazonium salt is coupled to N1 -napthylethylenediamine to form a diazoamino of a red colour.
  • the assay can be performed with different sensitivities, according to the grade of contamination to be analyzed.
  • several test strips have been elaborated to read the minimal acceptable concentration of endotoxin in aqueous solution (Table 1 ).
  • Figure 4 illustrates the resulting absorbance due to the - 12 - variation of the LAL which allow 4 classes of test-strips. Applied to the test-strip of example 2, it is possible to prepare a test-strip especially for the detection of endotoxin concentration in dialysate-solution, for which the acceptable limit is prescribed to be below 0.5 EU/ml. Detection spots for this prepared test strip are illustrated in Figure 5.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Urology & Nephrology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Hematology (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
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  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)

Abstract

L'invention concerne un appareil à phase solide permettant de détecter l'endotoxine dans un échantillon liquide, à l'aide de réactifs situés à l'intérieur d'un support solide, cet appareil comprenant un réactif sensible à l'endotoxine ainsi que des composants tampons. Lorsque l'on applique à cet appareil un échantillon liquide contenant ladite endotoxine, le réactif sensible à l'endotoxine réagit avec cette dernière de manière à former au moins un produit, le contact de ce produit et desdits composants tampons permettant par ailleurs de former un indicateur d'endotoxine. L'invention concerne également une méthode et un kit d'essai permettant de détecter l'endotoxine.
PCT/GB1999/001099 1998-04-09 1999-04-09 Essai en phase solide pour detecter l'endotoxine WO1999053322A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU34328/99A AU3432899A (en) 1998-04-09 1999-04-09 Solid phase test for endotoxin
CA002327169A CA2327169A1 (fr) 1998-04-09 1999-04-09 Essai en phase solide pour detecter l'endotoxine
EP99915905A EP1042672A1 (fr) 1998-04-09 1999-04-09 Essai en phase solide pour detecter l'endotoxine

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9807814.0 1998-04-09
GBGB9807814.0A GB9807814D0 (en) 1998-04-09 1998-04-09 Solid phase test for endoxin

Publications (1)

Publication Number Publication Date
WO1999053322A1 true WO1999053322A1 (fr) 1999-10-21

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Application Number Title Priority Date Filing Date
PCT/GB1999/001099 WO1999053322A1 (fr) 1998-04-09 1999-04-09 Essai en phase solide pour detecter l'endotoxine

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EP (1) EP1042672A1 (fr)
AU (1) AU3432899A (fr)
CA (1) CA2327169A1 (fr)
GB (1) GB9807814D0 (fr)
WO (1) WO1999053322A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005010207A2 (fr) 2003-03-17 2005-02-03 Charles River Laboratories, Inc. Procedes et compositions de detection de contaminants microbiens
US7479375B2 (en) 2005-01-13 2009-01-20 Charles River Laboratories, Inc. Method for classifying a microorganism in a biological sample
US7968280B2 (en) 2004-12-02 2011-06-28 Charles River Laboratories, Inc. Methods for the detection and/or quantification of gram positive bacterial contaminants
US8142735B2 (en) * 2001-10-29 2012-03-27 Arkray, Inc. Test apparatus
US20130078665A1 (en) * 2012-07-18 2013-03-28 Deepika Bodapati Test strip, a test kit and a method for detection of endotoxin in food

Citations (7)

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Publication number Priority date Publication date Assignee Title
US4717658A (en) * 1985-12-03 1988-01-05 Miles Inc. Gram negative bacteria screening method with horseshoe crab amebocyte lysate (LAL)
EP0265127A1 (fr) * 1986-10-09 1988-04-27 National Research Development Corporation Essai d'endotoxine
EP0291856A2 (fr) * 1987-05-20 1988-11-23 Leif Baek Méthode pour la détermination de la présence d'endotoxine dans un échantillon
EP0588303A1 (fr) * 1992-09-14 1994-03-23 Seikagaku Kogyo Kabushiki Kaisha (Seikagaku Corporation) Réactif pour un essai spécifique pour l'endotoxin
US5356778A (en) * 1990-07-13 1994-10-18 Board Of Regents, The University Of Texas Method for detection of gram-negative bacterial liposaccharides in biological fluids
US5591645A (en) * 1987-03-27 1997-01-07 Becton, Dickinson & Co. Solid phase chromatographic immunoassay
WO1997044665A1 (fr) * 1996-05-24 1997-11-27 The Trustees Of The University Of Pennsylvania Procedes et trousses d'identification de l'endotoxine

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4717658A (en) * 1985-12-03 1988-01-05 Miles Inc. Gram negative bacteria screening method with horseshoe crab amebocyte lysate (LAL)
EP0265127A1 (fr) * 1986-10-09 1988-04-27 National Research Development Corporation Essai d'endotoxine
US5591645A (en) * 1987-03-27 1997-01-07 Becton, Dickinson & Co. Solid phase chromatographic immunoassay
EP0291856A2 (fr) * 1987-05-20 1988-11-23 Leif Baek Méthode pour la détermination de la présence d'endotoxine dans un échantillon
US5356778A (en) * 1990-07-13 1994-10-18 Board Of Regents, The University Of Texas Method for detection of gram-negative bacterial liposaccharides in biological fluids
EP0588303A1 (fr) * 1992-09-14 1994-03-23 Seikagaku Kogyo Kabushiki Kaisha (Seikagaku Corporation) Réactif pour un essai spécifique pour l'endotoxin
WO1997044665A1 (fr) * 1996-05-24 1997-11-27 The Trustees Of The University Of Pennsylvania Procedes et trousses d'identification de l'endotoxine

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
GUILFOYLE D E ET AL: "Evaluation of a chromogenic procedure for use with the Limulus lysate assay of bacterial endotoxins in drug products.", JOURNAL OF PARENTERAL SCIENCE AND TECHNOLOGY, (1985 NOV-DEC) 39 (6) 233-6, XP002110748 *

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8142735B2 (en) * 2001-10-29 2012-03-27 Arkray, Inc. Test apparatus
WO2005010207A2 (fr) 2003-03-17 2005-02-03 Charles River Laboratories, Inc. Procedes et compositions de detection de contaminants microbiens
WO2005010207A3 (fr) * 2003-03-17 2005-06-09 Charles River Lab Inc Procedes et compositions de detection de contaminants microbiens
JP2007501020A (ja) * 2003-03-17 2007-01-25 チャールズ リバー ラボラトリーズ， インコーポレイテッド 微生物夾雑物の検出のための方法および組成物
US7329538B2 (en) 2003-03-17 2008-02-12 Charles River Laboratories, Inc. Methods and compositions for the detection of microbial contaminants
US7939291B2 (en) 2003-03-17 2011-05-10 Charles River Laboratories, Inc. Methods for the detection of microbial contaminants
US10119969B2 (en) 2003-03-17 2018-11-06 Charles River Laboratories, Inc. Compositions for the detection of microbial contaminants
US7968280B2 (en) 2004-12-02 2011-06-28 Charles River Laboratories, Inc. Methods for the detection and/or quantification of gram positive bacterial contaminants
US8440394B2 (en) 2004-12-02 2013-05-14 Charles River Laboratories, Inc. Methods for the detection and/or quantification of gram positive bacterial contaminants
US7479375B2 (en) 2005-01-13 2009-01-20 Charles River Laboratories, Inc. Method for classifying a microorganism in a biological sample
US7901899B1 (en) 2005-01-13 2011-03-08 Charles River Laboratories, Inc. Method for classifying a microorganism in a biological sample
US20130078665A1 (en) * 2012-07-18 2013-03-28 Deepika Bodapati Test strip, a test kit and a method for detection of endotoxin in food

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CA2327169A1 (fr) 1999-10-21
EP1042672A1 (fr) 2000-10-11
GB9807814D0 (en) 1998-06-10
AU3432899A (en) 1999-11-01

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