EP0528895A4 - Simplified solid-phase immunobead assay for detection of ciguatoxin and related polyethers - Google Patents
Simplified solid-phase immunobead assay for detection of ciguatoxin and related polyethersInfo
- Publication number
- EP0528895A4 EP0528895A4 EP19910909223 EP91909223A EP0528895A4 EP 0528895 A4 EP0528895 A4 EP 0528895A4 EP 19910909223 EP19910909223 EP 19910909223 EP 91909223 A EP91909223 A EP 91909223A EP 0528895 A4 EP0528895 A4 EP 0528895A4
- Authority
- EP
- European Patent Office
- Prior art keywords
- toxin
- recited
- support
- fish
- kit
- 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.)
- Withdrawn
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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
-
- 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
- G01N33/56961—Plant cells or fungi
Definitions
- the invention relates to the use of antibodies for the detection of ciguatoxin in fish.
- S-EIA simple stick enzyme immuno- assay
- Monoclonal antibodies (MAbs) : The method of Schrier et al. , Hybridoma Techniques. Cold Harbor Laboratory. Cold Spring, NY, 1980, incorporated herein by this reference, based on the original report of Kohler et al., "Continuous Cultures of Fused Cells Secreting Antibody of Predefined Specifi ⁇ city,” Nature. 156, 494-497 (1975), incorporated herein by this reference, was used.
- Hybridoma preparation for Mab-CTX was carried out using puri ⁇ fied CTX according to methods of Hokama et al.
- S-EIA Stick Enzyme Immunoassay
- Solid-phase Immunobead Assay The solid phase consisted of a paddle made of a bamboo stick coated with organic base solvent correction fluid supplied by Pentel of America, Ltd., Torrance CA 905034. Immunobead consisted of blue colored latex, 0.314 ⁇ in diameter, supplied by Seradyn, Inc. , Particle Technology Division Ind. , IN. Optimization of SPI Assay: Various parameters were examined to eliminate non-specific binding of colored latex to a coated bamboo paddle. Various concentrations of MAb-CTX with a constant suspension of colored latex were attempted to give the best specificity and sensitivity. Experiments were per ⁇ formed with colored latex alone in suspension (1% wt/wt in PBS buffer) .
- MAb-CTX concentra ⁇ tions ranging from 0.05, 0.075, 0.10, 0.13, 0.20, 0.45, 0.50, to 1.0 mg/ml were added to the 1% colored latex.
- the liquid-paper-coated sticks were examined as-is or were coated with a 1% wt/wt solution of Human Serum Albumin (HSA) , then examined as non-Me- OH-fixed or MeOH-fixed with the MAb-CTX-colored late.
- HSA Human Serum Albumin
- the immunobead was mixed thor ⁇ oughly before use.
- Solid-phase Immunoassay An inch-deep incision is made into the filet portion of the fish near the head region, as shown below.
- the coated paddle end is inserted into the incision and pushed up and down to touch the fish tissue.
- the paddle is removed, air-dried, and fixed quickly (1-3 seconds) with absolute methanol. After air-drying, the coated methanol-fixed end of the paddle is immersed into 0.5 ml of the immunobead color suspension. After 2, 5, and 10 minutes, the stick is examined and washed in saline. Any fish giving the stick a distinct colora ⁇ tion after 5 minutes is considered positive. If negative, immersion is continued up to 10 minutes.
- Table 1 shows the species, source and the test values given by the S-EIA and SPIA procedures for fish implicated in ciguatera poisoning. All samples were obtained from the Department of Health, State of Hawaii , except for two specimens (Antigua and Cali ⁇ fornia) . The S-EIA values were all in the rejection category and in complete agreement with the SPIA results . The majority of the toxic fishes were from the Big Island of Hawaii and associated with the Caranx sp.
- Caranx sp. 117 110 (Ulua, Eapio) 2. Seriola dumerili 9 9 (Kahala, Jack)
- Bodianus sp. 4 2 (Aawa, Wrasse)
- Table 3 summarizes the comparison of S-EIA and SPIA of the two studies, totalling 436 fishes . There is an overall 80% agreement, with 20% dis ⁇ agreement (12% - -S-EIA/+SPIA, and 8% +S-EIA/-SSPIA) . The 20% samples of fish in categ ⁇ ories -/+ and +/- will be further examined follow ⁇ ing extraction and then chromatographed by silica gel , with final assessment of the fractions by guinea pig and mouse bioassay.
- Table 4 shows the preliminary data obtained by a voluntary sportsfisherman assessing the solid- phase immunoassay presented earlier.
- Mr. Dale Takata of the Honolulu City Water and Waste management is summarized.
- Mr. Takata volun ⁇ teered to use the new SPIA test Of the 55 fishes of various species caught, 37 negative and border ⁇ line fishes were eaten, and 18 were rejected as being positive. Consumption of two fishes (papio and kole) from the positive group caused ciguatera poisoning in two individuals who were warned of the fish's toxicity based on the SPIA test.
- the SPIA method described should be applicable to other antigen-antibody systems, espe ⁇ cially when the antigen is an epitope or hapten of low molecular weight.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Virology (AREA)
- Peptides Or Proteins (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US51131790A | 1990-04-20 | 1990-04-20 | |
US511317 | 1990-04-20 |
Publications (2)
Publication Number | Publication Date |
---|---|
EP0528895A1 EP0528895A1 (en) | 1993-03-03 |
EP0528895A4 true EP0528895A4 (en) | 1993-03-10 |
Family
ID=24034379
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP19910909223 Withdrawn EP0528895A4 (en) | 1990-04-20 | 1991-04-19 | Simplified solid-phase immunobead assay for detection of ciguatoxin and related polyethers |
Country Status (6)
Country | Link |
---|---|
EP (1) | EP0528895A4 (en) |
JP (1) | JPH05507554A (en) |
AU (1) | AU647125B2 (en) |
BR (1) | BR9106366A (en) |
CA (1) | CA2080925A1 (en) |
WO (1) | WO1991016631A1 (en) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5286498A (en) * | 1992-05-01 | 1994-02-15 | Hawaii Chemtect Incorporated | Rapid extraction of ciguatoxin from contaminated tissues |
CN101963614A (en) * | 2010-09-03 | 2011-02-02 | 青岛科技大学 | The capillary electrophoresis electrochemical enzyme-linked immuno assay detects the method for ciguatoxin |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4703017C1 (en) * | 1984-02-14 | 2001-12-04 | Becton Dickinson Co | Solid phase assay with visual readout |
US4816392A (en) * | 1984-10-02 | 1989-03-28 | Research Corporation Of The University Of Hawaii | Rapid stick test for detection of ciguatoxin and other polyether toxins from tissues |
-
1991
- 1991-04-19 WO PCT/US1991/002703 patent/WO1991016631A1/en not_active Application Discontinuation
- 1991-04-19 JP JP91508651A patent/JPH05507554A/en active Pending
- 1991-04-19 BR BR919106366A patent/BR9106366A/en not_active Application Discontinuation
- 1991-04-19 EP EP19910909223 patent/EP0528895A4/en not_active Withdrawn
- 1991-04-19 AU AU77891/91A patent/AU647125B2/en not_active Ceased
- 1991-04-19 CA CA 2080925 patent/CA2080925A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
AU7789191A (en) | 1991-11-11 |
WO1991016631A1 (en) | 1991-10-31 |
AU647125B2 (en) | 1994-03-17 |
EP0528895A1 (en) | 1993-03-03 |
JPH05507554A (en) | 1993-10-28 |
BR9106366A (en) | 1993-04-27 |
CA2080925A1 (en) | 1991-10-21 |
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Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 19921023 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AT BE CH DE DK ES FR GB GR IT LI LU NL SE |
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A4 | Supplementary search report drawn up and despatched |
Effective date: 19930119 |
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AK | Designated contracting states |
Kind code of ref document: A4 Designated state(s): AT BE CH DE DK ES FR GB GR IT LI LU NL SE |
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17Q | First examination report despatched |
Effective date: 19950102 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 19950513 |