Cigua-dart method for detection of ciguatera toxins

• US 6,652,807 B1
• Filed: 07/10/2001
• Issued: 11/25/2003
• Est. Priority Date: 07/13/2000
• Status: Expired due to Fees

• Current Assignee: Oceanit Test Systems, Inc.
• Original Assignee: Oceanit Test Systems, Inc.
• Inventors: Pernambuco-Wise, Paul, Ebesu, Joanne S. M.
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): Counts, Gary W.
• Application Number:
  US09/901,016
• Time in Patent Office: 868 Days
• Field of Search: 422/58, 422/100, 422/101, 422/102, 422/103, 422/104, 435/808, 435/283.1, 435/287.2, 435/288.7, 436/165, 436/166, 436/174, 436/175, 436/177, 436/180, 600/562, 600/564-567
• US Class Current: 422/408
• CPC Class Codes: G01N 1/08 involving an extracting tool, e.g. core bit
• CPC Class Codes: G01N 21/658 enhancement Raman, e.g. surface plasmons
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10T 436/25125 Digestion or removing interfering materials

Abstract

A device for rapidly collecting tissue samples from organisms serves as a reaction chamber. It includes a solvent used to extract the analyte, a filter, and optional bioconcentrators. The filtered suspension is irradiated with a predetermined wavelength and the emitted, scattered, or reflected photons transmitted to a detector that identifies and quantifies the analyte. The bioconcentrators may consist of antibodies and colloidal metal nanoparticles that enhance emission of Raman signal frequencies by analytes bound to the antibodies. Alternatively, the device may contain only the extraction solvent and a filter.

Claims

1. Toxin detection apparatus comprising a housing having a main chamber for receiving a tissue sample, first and second end chambers having areas smaller than an area of the main chamber and communicating with opposite ends of the main chamber, a removable reagent cap for coupling with the first end chamber, a reagent in the reagent cap for expressing on the tissue sample in the chambers, an assay in the second end chamber for assaying substances leached from the tissue sample, and an analyzer for receiving data from the assay and analyzing substances in the tissue sample for detecting toxins.

2. The apparatus of claim 1, wherein the first end chamber comprise an open-ended tissue coring tube for collecting tissue sample.

3. The apparatus of claim 2, wherein the second end chamber comprise a closed-ended assay tube forming the assay for analyzing the tissue sample.

4. The apparatus of claim 3, wherein the reagent cap comprises first and second squeezable reagent caps interchangeably coupled to the coring tube for pushing the tissue sample and the first and second reagents respectively into the main chamber.

5. The apparatus of claim 4, wherein the first reagent in the first reagent cap comprises a solvent and the second reagent in the second reagent cap comprises a suspension comprising silver colloid particles and antibody specific for analytes of interest in the tissue sample.

6. The apparatus of claim 5, further comprising a solution formed by the reagent and the analytes and a filter for filtering the solution into the assay tube for analysis.

7. The apparatus of claim 6, wherein the filter comprises a filtration medium.

8. The apparatus of claim 6, further comprising a protective cap for covering the second end chamber.

10. The apparatus of claim 1, further comprising connectors on the first end chamber and fasteners on the reagent cap for removably connecting the reagent cap to the first end chamber.

11. The apparatus of claim 10, wherein the connectors and fasteners are threads.

12. The apparatus of claim 4, wherein each reagent cap comprises a hollow chamber for containing the reagents, and an open-ended hollow tube connecting the hollow chamber and the main chamber through the coring tube when the reagent cap is coupled to the first end chamber, wherein the first or the second squeezable reagent cap expresses the reagent from the hollow chamber into the main chamber on squeezing.

13. The apparatus of claim 12, further comprising a thin rupturable membrane between the hollow chamber and the hollow tube for rupturing and expressing the reagents.

15. The apparatus of claim 3, further comprising an energy source for supplying optical energy to the assay and wherein the analyzer is a detector for detecting optical energy from the assay.

16. The apparatus of claim 15, wherein the optical energy comprises radiation of varying wavelengths and intensities, and wherein the assay tube reflects, shifts or scatters the optical energy for detecting with the detector.

17. The apparatus of claim 16, wherein the detector comprises a charge-coupled device.

18. The apparatus of claim 16, further comprising a computing device for analyzing data collected by the detector, comparing analyzed data with data of control samples, quantifying and outputting analysis results corresponding to detected toxins.

19. The apparatus of claim 1, wherein the tissue sample is a seafood tissue sample.

20. The apparatus of claim 1, wherein the tissue sample is a fish tissue sample.

9. Toxin detection apparatus comprising a main chamber for receiving a tissue sample, first and second end chambers on opposite ends of the main chamber, a removable reagent cap for coupling with the first end chamber, a reagent in the reagent cap for expressing on the tissue sample in the chambers, an assay in the second end chamber for assaying substances leached from the tissue sample, a detector for receiving data from the assay, analyzing and detecting toxins, and the first end chamber forming an open-ended tissue coring tube, wherein the tissue coring tube is beveled to facilitate sample collection.

14. Toxin detection apparatus comprising a main chamber for receiving a tissue sample, first and second end chambers on opposite ends of the main chamber, a removable reagent cap for coupling with the first end chamber, a reagent in the reagent cap for expressing on the tissue sample in the chambers, an assay in the second end chamber for assaying substances leached from the tissue sample, a detector for receiving data from the assay, analyzing and detecting toxins, and wherein the first end chamber comprises an open-ended tissue coring tube for collecting tissue sample, wherein the tissue coring tube further comprises bevelled tips.