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Capillary immunity-chromatography rapid detection method for parvalbumin in aquatic products

A technology of immunochromatography and detection method, which is applied in the detection field of parvalbumin, can solve the problems such as poor results, and achieve the effects of improving detection accuracy, facilitating storage, and reducing the effect of cross-binding

Active Publication Date: 2015-06-17
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Parvalbumin is an acidic protein bound to calcium ions. It is relatively stable to heat, protease hydrolysis and chemical denaturation. Therefore, inactivation and removal methods do not work well. Establish a sensitive, reliable and rapid parvalbumin Protein detection methods become the focus of attention

Method used

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  • Capillary immunity-chromatography rapid detection method for parvalbumin in aquatic products
  • Capillary immunity-chromatography rapid detection method for parvalbumin in aquatic products
  • Capillary immunity-chromatography rapid detection method for parvalbumin in aquatic products

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preparation example Construction

[0068] Preparation of anhydrous toluene: Add 15 g of anhydrous sodium sulfate to 300 mL of toluene (analytical pure) and let it stand for 10 hours, remove the anhydrous sodium sulfate by suction filtration, then add 2 g of metallic sodium wire, and add 0.5 g of benzophenone as an indicator , replace the air in the reflux device with nitrogen, and heat to reflux for 2 hours to make the solution turn blue; stop heating, change to a distillation device, and then distill at atmospheric pressure to collect the distillate at 111±1°C to obtain anhydrous toluene.

[0069] Dissolve GPTMS and triethylamine in anhydrous toluene to obtain a modification solution, so that the final concentration of GPTMS is 8 vol%, and the final concentration of triethylamine is 1 vol%, and immerse the capillary obtained in step (1) in the modification solution at room temperature React in a dry environment for 25 hours, drain the modification solution in the capillary, keep it in a dry environment at room ...

Embodiment 1

[0104] 1. Treatment of samples to be tested: Mix fish (turboscus purchased from Jusco Supermarket (Qingdao)) and Tris-HCl (pH 7.5) at a ratio of 1:2 (w / v) to homogenate, and homogenate After the final sample was filtered, it was heated in a water bath at 95°C for 25 minutes, centrifuged at 3800g for 5 minutes, and the supernatant (extracted parvalbumin) was collected as a test sample.

[0105] 2. Sample detection: Mix the colloidal gold-labeled primary antibody prepared in Section 4.1 (gold-labeled rabbit anti-parvalbumin) with the test sample at a volume ratio of 1:1 to obtain a mixture, and inject 5 μL of the mixture into the immunochromatographic capillary After standing still for 4 minutes, use a pipette to push the mixed solution to move down to the quality control area of ​​the immunochromatography capillary, and also stay in this area for 4 minutes, drain the excess mixed solution out of the immunochromatography capillary, and then The immunochromatographic capillary wa...

Embodiment 2

[0107] 1. Treatment of samples to be tested: Mix fish (turboscus purchased from Jusco Supermarket (Qingdao)) and Tris-HCl (pH 7.5) at a ratio of 1:3 (w / v) to homogenate, and homogenate After the final sample was filtered, it was heated in a water bath at 100°C for 5 minutes, centrifuged at 3800g for 5 minutes, and the supernatant (extracted parvalbumin) was collected as a test sample.

[0108] 2. Sample detection: Mix the colloidal gold-labeled primary antibody prepared in Section 4.1 (gold-labeled rabbit anti-parvalbumin) with the test sample at a volume ratio of 1:1.5 to obtain a mixture, and inject 15 μL of the mixture into the immunochromatographic capillary After standing still for 4 minutes, use a pipette to push the mixed solution to move down to the quality control area of ​​the immunochromatography capillary, and also stay in this area for 4 minutes, drain the excess mixed solution out of the immunochromatography capillary, and then The immunochromatographic capillary...

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Abstract

A disclosed capillary immunity-chromatography rapid detection method for parvalbumin in aquatic products comprises the following steps: 1, preparing a to-be detected sample, specifically mixing fish flesh with Tris-HCl, homogenizing, filtering, heating in a water bath, centrifuging, and colleting a supernatant as a to-be detect sample; and 2, detecting the sample, specifically mixing uniformly a first antibody labeled by colloidal gold with the to-be detected sample to obtain a mixed solution, fetching 5-15 mu L of the mixed solution, injecting into the end of a detection zone of an immunity-chromatography capillary, standing for 4 min, pushing the mixed solution to flow downwards to a quality-control zone of the immunity-chromatography capillary by using a pipettor, also staying for 4 min at the zone, discharging excessive solution out of the capillary, then immersing the immunity-chromatography capillary in a PBST buffer for twitch cleaning, and finally obtaining a detection result through naked-eye qualification. The method is good in detection sensitivity, high in repeatability, short in detection time and good instability, and is applicable to instant rapid detection.

Description

technical field [0001] The invention relates to a detection method of parvalbumin, in particular to a capillary immunochromatographic detection method of parvalbumin in aquatic products. Background technique [0002] With the rapid and stable growth of my country's economy, people's quality of life continues to improve, and the requirements for food consumption are also getting higher and higher. However, environmental pollution, residues of pesticides and veterinary drugs, food allergens and other incidents that endanger food safety have been reported continuously and have attracted widespread attention. [0003] In the early 1980s, immunocolloidal gold chromatography technology was widely used in the fields of medicine, environment, food testing, agriculture and animal husbandry due to its high sensitivity, strong specificity, simple operation, and no need for instruments. The core technology of the colloidal gold immunochromatography method is to use the strip-shaped fib...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/12G01N33/68
Inventor 林洪杜淑媛曹立民隋建新王静雪
Owner OCEAN UNIV OF CHINA
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