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Quick detection kit for okadaic acid toxins in shellfish

A technology of okadaic acid and a kit, which is applied in the field of rapid detection kits for okadaic acid toxoids, can solve the problems of difficult on-site, rapid detection, cumbersome detection methods, expensive instruments, etc., and achieve simplified sample pretreatment and detection steps, avoiding ethical issues, and easy operation

Inactive Publication Date: 2014-10-08
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem solved by the present invention is to provide a rapid detection kit for okadaic acid toxoid in shellfish to overcome the defects of existing detection methods such as cumbersome detection methods, time-consuming detection, expensive instruments, and difficulty in realizing on-site and rapid detection.

Method used

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  • Quick detection kit for okadaic acid toxins in shellfish
  • Quick detection kit for okadaic acid toxins in shellfish
  • Quick detection kit for okadaic acid toxins in shellfish

Examples

Experimental program
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Embodiment 1

[0028] A rapid detection kit for okadaic acid toxoid in shellfish, specifically comprising:

[0029] (1) 96-well microwell plate (Corning, USA); it can be disassembled into 12 microwell strips, which can test multiple samples at one time, and can also be disassembled for separate testing. The remaining microwell strips can be stored in a ziplock bag, Use it next time.

[0030] (2) Protein phosphatase 2A freeze-dried powder (Zeus Biotechnology Company, Spain), 2.5U / cartridge, 5 bottles in total; each enzyme freeze-dried powder was prepared with 2mL protein phosphatase 2A dilution solution before use, and prepared at room temperature ( Slightly oscillate (rotating speed <60rpm) on an enzyme label shaker (22±2°C) for 60±5min to fully hydrolyze it, and use it immediately after preparation.

[0031] (3) Concentration gradient series standard solution of okadaic acid; a total of 6 bottles of standard solution series with gradient concentration of okadaic acid, the concentrations ar...

Embodiment 2

[0037] The method for the total toxic equivalent of applying the test kit of the present invention to measure OA toxoid may further comprise the steps:

[0038] (1) Preparation of sample solution: Weigh 2.00 g of homogenized shellfish sample, add 10 mL of methanol, extract homogeneously at 10,000 r / min for 2 min, centrifuge at 4,000 r / min for 10 min, then pipette 1 mL of methanol extract into a 5 mL centrifuge tube, first Add 125μL 2.5mol / L NaOH, vortex for 10s, heat in a water bath at 76±2℃ for 40min, then add 125μL 2.5mol / L HCl to neutralize. Finally, blow the solution nearly dry with nitrogen, add buffer solution (pH 8.4) to make the volume to 20mL, and wait for the test.

[0039] (2) Preparation of protein phosphatase 2A (PP2A enzyme) solution: select the amount of enzyme according to the number of samples. Add 2 mL of enzyme dilution solution to each enzyme lyophilized powder, and shake slightly (rotating speed <60 rpm) on an enzyme label shaker at room temperature (22±2...

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Abstract

The invention discloses a quick detection kit for okadaic acid toxins in shellfish, belonging to the technical field of food detection. The quick detection kit comprises (1) protein phosphatase 2A freeze-dried powder, (2) an okadaic acid concentration gradient series standard solution, (3) a buffer solution, (4) a protein phosphatase 2A dilute solution, (5) a substrate developing solution, and (6) a high-concentration marking solution. According to the technical scheme disclosed by the invention, the quick detection kit inherits the advantage that a mouse bioassay can establish a dosage-effect relation, so that the relative toxicity of the toxins can be directly reflected; compared with the mouse bioassay, the quick detection kit has the advantage that a large batch of samples (84 samples can be detected within 4 hours) can be detected within short time. The detection limit of the method disclosed by the invention is 50 microgram OA eq. / kg shellfish tissue; compared with the detection limit, 200 microgram OA eq. / kg shellfish tissue, of the conventional mouse bioassay, the detection limit of the method disclosed by the invention is greatly reduced; meanwhile, the ethic problem of an animal experiment is solved. The kit disclosed by the invention is low in cost and easy to operate.

Description

technical field [0001] The invention belongs to the technical field of food detection, and in particular relates to a rapid detection kit for okadaic acid toxoid in shellfish. Background technique [0002] Shellfish is loved by consumers for its delicious taste and rich nutrition, and it has become the leading species in my country's fishery industry. However, the accumulation of toxins in shellfish has plagued the healthy development of my country's shellfish industry for many years. In recent years, surveys of shellfish production areas in some sea areas in my country have shown that shellfish toxins have been found in Liaodong Bay, Laizhou Bay, Qinhuangdao, Fujian and other sea areas, and okadaic acid (OA) toxoids are more common. OA toxoid is a polycyclic polyether fat-soluble shellfish toxin, which is thermally stable and can remain in shellfish tissue for a long time; and conventional processing methods such as heating and microwaves reduce the water content in aquatic...

Claims

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Application Information

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IPC IPC(8): G01N21/78
Inventor 郭萌萌谭志军李兆新王智吴海燕赵春霞翟毓秀
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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