Method and box for quick enzyme immunity detecting food-borne parasitic disease

A parasitic disease and detection method technology, applied in the direction of measuring devices, analysis by making materials undergo chemical reactions, instruments, etc., can solve problems such as parasite ovulation, economic loss, irregular ovulation, etc., and achieve a high degree of consistency and comparability, avoiding hazards and pollution, and eliminating potential safety hazards

Inactive Publication Date: 2008-06-25
SHENZHEN COMBINED BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are two types of detection methods for foodborne parasitic diseases: one is the traditional detection method, that is, to find pathogens, because these four kinds of parasites are mostly parasitic in deep tissues, such as brain, eyeball, liver, biliary tract, lung For viscera such as the body, skeletal muscle, and cardiac muscle, it is necessary to collect the worms through surgical means, which not only causes great damage, but also often makes the worms scattered and difficult to detect.
In addition, trichinella spiralis and cysticercosis do not ovulate eggs, and paragonimia seldom ovulates or even does not ovulate; although liver fluke can ovulate with feces, the eggs are small in size and irregular in ovulation, so it is difficult to detect them. The commonly used feces smear method in clinical Sensitivity generally below 50%
Therefore, no matter checking worm eggs or worm bodies, it is not an effective means for diagnosing this group of parasitic diseases; another type is the conventional enzyme immunoassay that has appeared in recent years, but these methods generally have the following shortcomings: 1. The operation time is long: The time is usually more than 2-4 hours, and it is difficult to meet the needs of rapid diagnosis in the field and clinical outpatient clinics; The termination liquid needs to be prepared by the user, which brings a lot of inconvenience to the user in the application; 3. Instability: the provided reagent is not stable, and once prepared, it must be used up within 1 week or even 1 day, resulting in economic losses. It also affects the reliability of detection; 4. Incomplete supporting facilities: so far, there is no complete kit for four food-borne parasites with a unified method in the market, and all kits can only detect one parasitic disease
Different manufacturers, different methods, and different performance not only affect the operability, but also affect the consistency and effectiveness of the results, which may easily cause misdiagnosis or missed diagnosis, and become a major hidden danger in the prevention and control of foodborne parasitic diseases; 5 Safety Hidden dangers: conventional enzyme immunoassays often use teratogenic OPD as a chromogenic agent, and use highly corrosive sulfuric acid as a stop solution, which poses a threat to operators and the environment
In summary, the existing detection technology cannot effectively meet the needs of the diagnosis of foodborne parasitic diseases, which is not conducive to the diagnosis, treatment, prevention and control of the disease.

Method used

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  • Method and box for quick enzyme immunity detecting food-borne parasitic disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Rapid enzyme immunoassay method for liver fluke disease

[0017] The soluble crude antigen of liver fluke was selected, and the purified antigen was purified step by step by the octanoic acid method and DEAE52 ion chromatography column chromatography, and the protein content was determined by the ultraviolet method; the preliminary test was carried out, and the contrast experiment with the other three antigens was determined and selected. Coating concentration of trematode refined antigen; dilute antigen with carbonate buffer (composed of sodium bicarbonate and sodium carbonate, pH9.60, 0.05M), coat 96-well polystyrene microwell plate, 100 microliters / well , overnight at room temperature, shake off the liquid in the well, and block with the same dose of 1% bovine serum albumin-containing phosphate buffer (composed of sodium dihydrogen phosphate and disodium hydrogen phosphate, pH7.0, 0.05M) at room temperature for 2 hours , Freeze and vacuum dry, put into plas...

Embodiment 2

[0022] Example 2 Rapid Enzyme Immunoassay Detection Method for Paragonimiasis

[0023] The soluble crude antigen of lung fluke was selected, and the purified antigen was purified step by step by octanoic acid method and DEAE52 ion chromatography column chromatography. Fluke refined antigen coating concentration; dilute antigen with carbonate buffer (composition as above, PH9.60, 0.05M), coat 96-well polystyrene microplate, 100 microliters / well, overnight at room temperature, shake off The liquid in the well was closed at room temperature for 2 hours with the same dose of phosphate buffer containing 1% bovine serum albumin (composition as above, PH7.0, 0.05M), freeze-dried in vacuum, bagged under vacuum conditions, and placed on the plate bag Label accordingly. The coated board can also be used directly after unpacking.

[0024] Other reagent preparation methods, assembly into boxes and detection schemes are the same as in Example 1.

Embodiment 3

[0025] Embodiment 3 swine cysticercosis rapid enzyme immunoassay method

[0026] The soluble crude antigen of porcine cyst fluid was selected, and the purified antigen was purified step by step by octanoic acid method and DEAE52 ion chromatography column chromatography, and the protein content was determined by ultraviolet method; the preliminary test was carried out, and the comparison experiment with the other three antigens was determined. And select the antigen coating concentration; dilute the antigen with carbonate buffer (composition as above, PH9.60, 0.05M), coat 96-well polystyrene microwell plate, 100 microliters / well, overnight at room temperature, shake off The liquid in the well was closed at room temperature for 2 hours with the same dose of phosphate buffer containing 1% bovine serum albumin (composition as above, PH7.0, 0.05M), freeze-dried in vacuum, bagged under vacuum conditions, and placed on the plate bag Label accordingly. The coated board can also be us...

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Abstract

The invention discloses a food-borne verminosis fast enzyme immunity detection method and a perforated detection kit, wherein, an enzyme immunity detection reagent comprises an antigen coating plate A, enzyme conjugate liquid B, cleaning solution C, a substrate D, a color developing reagent E, sample diluent F, terminator G, a positive reference substance H and a negative reference substance I; the perforated detection kit is used during the detection process which comprises sample dilution, sample adding reaction, enzyme adding reaction and color developing reaction; the antigen coating plate A adopts quick coating technology, and antigens are coated on a general purpose polystyrene mipor plate in advance; two polyethylene glycols with molecular weight Mr of 20000 and 6000 are added into the enzyme conjugate liquid B. The invention solves the problems of the prior detection method like long operating time, trivial operation, instable reagents, incomplete match and so on, and has the advantages of quick speed, convenient operation, high reliability, good detection results and so on. The invention is used for enzyme immunity detection of cysticercosis cutis, paragonimiasis, distomatosis and trichinosis.

Description

technical field [0001] The invention relates to a rapid enzyme immunological detection method and detection box for food-borne parasitic diseases. Background technique [0002] As we all know, food-borne parasitic diseases (FOOD SOURCE PARASITEDISEASES) are a group of common parasitic diseases that can be transmitted through food, mainly including four parasitic diseases of pig cysticercosis, lung fluke, liver fluke and trichinella spiralis. They can infect humans as well as domestic animals, causing serious damage to human health and economic losses in the livestock industry. Among them, Cysticercosis suis can cause epilepsy, paralysis, and blindness; lung fluke can cause lung cysts and lung abscesses; liver fluke can cause biliary tract infection, gallstones, and liver cirrhosis; trichinella can cause severe allergic reactions and heart damage. Foodborne parasites are widespread, and the population of liver fluke endemic areas in Guangdong Province, China alone has reache...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/531G01N21/78
Inventor 杜宜峰
Owner SHENZHEN COMBINED BIOTECH
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