Enzyme-linked immunosorbent assay kit suitable for pentachlorophenol residual analysis

An enzyme-linked immunosorbent assay and detection kit technology, applied in analytical materials, biological tests, measuring devices, etc., can solve problems such as environmental pollution, high requirements for instrumentation, slow analysis speed, etc. Less time consuming effect

Inactive Publication Date: 2009-09-09
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Applying these physical and chemical analysis techniques to analyze trace amounts of pentachlorophenol residues in environmental, biological, food and other samples not only requires a high degree of instrumentation, but also requires complicated pretreatment processes such as separation, extraction, purification, and derivation. Slow, high cost, the pre-treatment process requires the use of

Method used

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  • Enzyme-linked immunosorbent assay kit suitable for pentachlorophenol residual analysis
  • Enzyme-linked immunosorbent assay kit suitable for pentachlorophenol residual analysis

Examples

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Embodiment 1

[0017] Example 1 Kit operation and result calculation

[0018] After the samples to be tested were pretreated, they were made to volume with PBST for later use. Unpack the vacuum packaging bag and take out the microtiter plate, and equilibrate at room temperature for 5 minutes for later use. Prepare 0ng·mL -1 , 0.064ng·mL -1 , 0.32ng·mL -1 , 1.6ng·mL -1 , 8ng·mL -1 , 40ng·mL -1 , 200ng·mL -1 , 1000ng·mL -1 Pentachlorophenol standard solution, add 50 μL standard sample or processed sample to each well, standard sample and sample are repeated 4 times, add 50 μL diluted antibody, and incubate at 37°C for 30 minutes; pour out the liquid in the well, and use Wash the diluted PBST 5 times, invert the ELISA plate on absorbent paper and pat dry; add 100 μL of ELISA-labeled goat anti-rabbit secondary antibody diluted at 1:1000, and incubate at 37°C for 30 minutes; pour out the liquid in the well, Wash the plate 5 times with PBST and pat it dry; mix equal volumes of liquid A an...

Embodiment 2

[0020] Example 2 Synthesis of hapten tetrachloro-p-hydroxyphenoxybutyric acid

[0021] Add about 100mL of acetone solvent to a 250mL round-bottomed three-neck flask, under nitrogen protection, add 3g (12.15mmol) tetrachlorohydrodiphenol and 10g (72.5mmol) potassium carbonate, stir to dissolve as much as possible, and dissolve 1.58g (8.1mmol) Ethyl 4-bromobutyrate was dissolved in a small amount of acetone, added dropwise to the reaction solution, and started to reflux for about 12 hours.

[0022] Thin-layer liquid chromatography (TLC) detected that the reaction was complete, and the acetone solvent was removed under reduced pressure. The reactants were dissolved in 50 mL of deionized water, and then 30 mL of ethyl acetate was added, fully shaken, allowed to stand and separated, and the water phase was retained. Add dilute hydrochloric acid solution to the water phase to adjust the pH to 3, extract the water phase with ethyl acetate 50mL×2, combine the organic phase, wash the ...

Embodiment 3

[0024] Example 3 Preparation of polyclonal antibody against pentachlorophenol

[0025] The hapten tetrachloro-p-hydroxyphenoxybutyric acid was coupled with bovine serum albumin by the active ester method, and 2 mg of the conjugate was dissolved in 1 mL of normal saline, mixed with 1 mL of complete Freund's adjuvant, fully emulsified, and then injected into New Zealand Thighs of white-eared rabbits were given a booster immunization once every two weeks, and mixed with incomplete Freund's adjuvant and the immunogen. The immunization site was subcutaneous on the back of the neck. Starting from the third immunization, one week after each immunization, it was injected from the rabbit ear vein Blood was collected to test the serum titer. A total of 5 times of immunization, the whole blood was collected from the rabbit carotid artery one week after the last immunization, and the rabbit antiserum was crudely extracted by 35% saturated ammonium sulfate salting out method, and finally p...

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Abstract

The invention provides an enzyme-linked immunosorbent assay kit suitable for pentachlorophenol residual analysis, which comprises elisa plate coated with pentachlorophenol antigen, concentrated cleaning liquid, pentachlorophenol standard, pentachlorophenol specific antibody, enzyme marker, coloration liquid and reaction stopping solution. The pentachlorophenol with the known concentration is detected and a standard curve is drawn, and then the concentration of the pentachlorophenol to be detected can be deduced. The invention has the advantages that the pentachlorophenol residual in the environment can be accurately and sensitively detected, the pretreatment process of the samples is simple and consumes less time, a great amount of the samples can be detected, and the detection cost of the samples is far lower than that of the detection method of the traditional instrument.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent assay (ELISA) kit, in particular to an enzyme-linked immunosorbent assay kit suitable for the analysis of pentachlorophenol residues. Background technique [0002] Pentachlorophenol has a wide range of uses. As a pesticide, it is mainly its sodium salt. It can be used as a fungicide, preservative, herbicide, and a wood preservative. Some of it is released into nature as a solvent and heat exchange industrial production waste. Environment. Pentachlorophenol has strong biological toxicity because of its biological uncoupling effect. The endocrine disrupting effect of pentachlorophenol is manifested in interfering with the normal thyroid hormone function of the body; interfering with (simulating) estrogen and testosterone, thereby affecting human fertility. my country's urban sewage treatment plant pollutant discharge standard GB 18918-2002 stipulates that the concentration of pentachlorophenol poll...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/53
Inventor 李季许艇井宏宇
Owner CHINA AGRI UNIV
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