CN201242547Y - Immune ratio turbidity quantitative determination kit for cow's milk immune globulin - Google Patents

Abstract

The utility model provides an immune turbidimetry quantitative detection kit for bovine milk immunoglobulins IgG, which relates to a detection kit. The utility model solves the problems that the time for detecting the IgG content of the immune globulin in the prior cow milk and the products thereof is long, and the detection accuracy is poor. A fixing plate of the utility model is arranged in a kit body; a first bottle hole, a second bottle hole, a third bottle hole and a fourth bottle hole are arranged on the fixing plate, an immune globulin reagent bottle is arranged in the first bottle hole, an immune globulin antibody reagent bottle is arranged in the second bottle hole, a phosphate buffer solution reagent bottle is arranged in the third bottle hole, and a carbowax reagent bottle is arranged in the fourth bottle hole. The utility model has the advantages that by utilizing the reagent in the immune globulin reagent bottle, the immune globulin antibody reagent bottle, the phosphate buffer solution reagent bottle and the carbowax reagent bottle, the detection kit can detect the IgG content of the immune globulin in the prior cow milk and the products thereof rapidly, the detection time is short, the detection is accurate, and the operation is simple and convenient.

Description

The immunoturbidimetry detection by quantitative kit of milk immune globulin IgG

Technical field

The utility model relates to a kind of detection kit.

Background technology

Immunoglobulin IgG is as a kind of very important immune factor in the colostrum, and its content and other bioactive substance content have certain positive correlation, its content how much be to weigh a good and bad important indicator of colostrum and products thereof.Immunoglobulin IgG also is the major function composition of immune milk and goods thereof, and its content is the most important standard of immune milk and quality of item quality thereof.Whether can also assess normal Ruzhong to the detection of milk immune globulin IgG is mixed with colostrum and judges whether cow's milk is mingled.Thereby the detection of milk immune globulin has very important meaning for the quality control of dairy products.

In order to guarantee the quality of colostrum, immune milk and normal dairy products, we often need be in each link of production quality control be strengthened newborn sample the accurate detection of immunoglobulin level.More above-mentioned different milk immune globulin IgG detection method, can find conventional immunologic detection method such as immune uni-directional diffusion, immune directional diffusion and rocket immunoelectrophoresis, these methods all have advantage easy and simple to handle, that equipment requirements is low, but exist the shortcoming of length detection time, poor accuracy.Though the colloidal gold immunochromatographimethod technology can fast detecting, is confined to qualitative or half-quantitative detection, can't reach the purpose of accurate detection.Though and high performance liquid chromatogram and Capillary Electrophoresis can be fast, accurately detect, to equipment and operating personnel require highly, be not suitable for conventional factory and conventional test chamber to analyze, popularize comparatively difficulty.

The utility model content

The purpose of this utility model is a problem long for the detection time that solves content of immunoglobulin IgG in existing cow's milk and the goods thereof, poor accuracy, and the immunoturbidimetry detection by quantitative kit of a kind of milk immune globulin IgG is provided.

The utility model is made up of immunoglobulin (Ig) reagent bottle, immune globulin antibody reagent bottle, phosphate buffer reagent bottle, polyethylene glycol reagents bottle, box body and fixed head, fixed head is arranged in the box body, fixed head is provided with first bottle of hole, second bottle of hole, the 3rd bottle of hole and the 4th bottle of hole, the immunoglobulin (Ig) reagent bottle is arranged in first bottle of hole, the immune globulin antibody reagent bottle is arranged in second bottle of hole, the phosphate buffer reagent bottle is arranged in the 3rd bottle of hole, and the polyethylene glycol reagents bottle is arranged in the 4th bottle of hole.

The utility model has the advantages that: the utility model utilizes the reagent in immunoglobulin (Ig) reagent bottle, immune globulin antibody reagent bottle, phosphate buffer reagent bottle and the polyethylene glycol reagents bottle, immunoglobulin content in cow's milk and the goods thereof is carried out fast quantification to be detected, detection time is short, detection is accurate, and easy and simple to handle, not high to equipment requirements, be more suitable for requirements of actual production.

Description of drawings

Fig. 1 is an one-piece construction synoptic diagram of the present utility model.

Embodiment

Embodiment one: present embodiment is described in conjunction with Fig. 1, present embodiment is by immunoglobulin (Ig) reagent bottle 1, immune globulin antibody reagent bottle 2, phosphate buffer reagent bottle 3, polyethylene glycol reagents bottle 4, box body 5 and fixed head 6 are formed, fixed head 6 is arranged in the box body 5, fixed head 6 is provided with first bottle of hole 6-1, second bottle of hole 6-2, the 3rd bottle of hole 6-3 and the 4th bottle of hole 6-4, immunoglobulin (Ig) reagent bottle 1 is arranged in first bottle of hole 6-1, immune globulin antibody reagent bottle 2 is arranged in second bottle of hole 6-2, phosphate buffer reagent bottle 3 is arranged in the 3rd bottle of hole 6-3, and polyethylene glycol reagents bottle 4 is arranged in the 4th bottle of hole 6-4.Immunoglobulin (Ig) reagent bottle 1 is brown plastic bottle, and the bottle inner sealing is being packed the Immunoglobulin IgG standard items by fixed weight and accurate weighing, and the quality of Immunoglobulin IgG standard items is 10mg, and this is reagent I, and 4 ℃ of preservations are used for the drafting of typical curve; Immune globulin antibody reagent bottle 2 is the white glass bottle, and the bottle inner sealing is being packed the Immunoglobulin IgG antibody powder of fixed weight, and this is reagent II, 4 ℃ of preservations; Phosphate buffer reagent bottle 3 is the white plastic bottle, the bottle inner sealing is being packed the phosphate buffer that is made into by fixed weight and proportioning, the concentration of phosphate buffer is 0.01mol/L, the pH value of phosphate buffer is 7.4, the phosphate buffer volume is 500mL, this is reagent III, 4 ℃ of preservations; Polyethylene glycol reagents bottle 4 is the white plastic bottle, and the bottle inner sealing is being packed the polyglycol by fixed weight and accurate weighing, and the quality of polyglycol is 15g, and this is reagent IV, 4 ℃ of preservations.Above-mentioned four reagent bottles are put into the first reagent bottle hole 6-1, the second reagent bottle hole 6-2, the 3rd reagent bottle hole 6-3 and the 4th reagent bottle hole 6-4 respectively, build with the lid 5-1 on the box body 5 and seal up.Reagent I, reagent II, reagent IV are solid, and reagent III is a liquid.

Embodiment two: in conjunction with Fig. 1 present embodiment is described, the material of the box body 5 of present embodiment is cardboard or plastic board.This material has in light weight, convenient carrying.

Embodiment three: in conjunction with Fig. 1 present embodiment is described, the material of the fixed head 6 of present embodiment is cardboard or plastic foam plate.This material has in light weight, convenient carrying.

Using method of the present utility model:

One, the preparation of reagent I, II, III, IV:

1, polyglycol is than turbid buffer preparation: take out reagent III from kit, join among the reagent IV, be mixed with 4% solution, be called polyglycol than turbid damping fluid, then 4 ℃ of preservations.

2, Immunoglobulin IgG standard stock solution preparation: take out reagent I from kit, add the reagent III of 10mL in reagent I, carry out mixing, its concentration is 1.0mg/mL, and it is standby to be placed in-20 ℃ of freezings after packing.

3, Immunoglobulin IgG standard serial solution preparation: get the above-mentioned Immunoglobulin IgG standard stock solution for preparing, be diluted to the Immunoglobulin IgG titer that concentration is respectively 0mg/mL, 0.1mg/mL, 0.2mg/mL, 0.3mg/mL, 0.4mg/mL, 0.5mg/mL, 0.6mg/mL, 0.7mg/mL with polyglycol than turbid damping fluid, face the time spent preparation.

4, Immunoglobulin IgG antibody stock solution preparation: reagent II antibody pulvis is dissolved among the reagent III of 2mL, and every pipe is distributed into 100 μ L, and it is standby to put-20 ℃ of freezings.

5, Immunoglobulin IgG antibody diluent: get the Immunoglobulin IgG antibody stock solution of 100 μ L, the polyglycol that adds 2.9mL with 30 times of antibody dilutions, faces the time spent preparation than turbid damping fluid.

Two, detect step:

1, the processing of sample to be checked:

(1) liquid newborn sample: get liquid cow's milk sample and carried out centrifugal 30 minutes, remove upper strata fat, measure 10mL skimmed milk sample then, be diluted to 1000～2000mL with polyglycol than turbid damping fluid, mixed evenly back is standby.

(2) solid-state newborn sample: after various types of solid-state cow's milk samples pulverizing, take by weighing 0.1g (being accurate to 0.001g) sample, adopt polyglycol to be diluted to 50～100mL than turbid damping fluid, mixing, standby (newborn sample to be measured is fat-containing component not); If newborn sample to be measured contains fat constituent, need dissolve earlier, centrifugal, remove fat, then operation is with (1).

2, the drafting of typical curve:

Get 8 reaction tubes, the Immunoglobulin IgG antibody diluent that in each reaction tube, adds 250 μ L, the Immunoglobulin IgG concentration of getting 10 μ L again is 0mg/mL, 0.1mg/mL, 0.2mg/mL, 0.3mg/mL, 0.4mg/mL, 0.5mg/mL, 0.6mg/mL, 0.70mg/mL standard serial solution add in each reaction tube, after mixing evenly, place 37 ℃ water-bath to react 40 minutes, after reaction finishes, with each tube reaction liquid on Biochemical Analyzer, the selection wavelength is 340nm, return to zero than turbid damping fluid with polyglycol, measure the optical density value of various criterion concentration Immunoglobulin IgG, the drawing standard curve.

3, the mensuration of sample to be checked:

In each reaction tube, add the Immunoglobulin IgG antibody diluent of 250 μ L, get 10 μ L testing samples again and add in each reaction tube, mixed evenly after, place 37 ℃ water-bath reaction 40 minutes.After reaction finished, on Biochemical Analyzer, the selection wavelength was 340nm, measures its optical density value with each tube reaction liquid, according to typical curve, calculates the content of the Immunoglobulin IgG in the testing sample.

4, blank assay:

The normal skimmed milk identical with testing sample concentration is blank sample, adopts identical determination step to carry out operation repetitive.

5, the result calculates:

$X=\frac{c\×V\×100}{m\×1000}$

In the formula:

The content of Immunoglobulin IgG in X-testing sample, unit is gram every hectogram (g/100g) or per hundred milliliters (mg/100mL) of milligram;

The quality of m-testing sample, unit is gram (g) or milliliter (mL);

The content of Immunoglobulin IgG in c-test solution, unit are every milliliter (mg/mL) of milligram;

The volume of V-testing sample constant volume, unit are milliliter (mL).

Result of calculation keeps two position effective digitals.

Claims (1)

1, the immunoturbidimetry detection by quantitative kit of a kind of milk immune globulin IgG, it is by immunoglobulin (Ig) reagent bottle (1), immune globulin antibody reagent bottle (2), phosphate buffer reagent bottle (3), polyethylene glycol reagents bottle (4), box body (5) and fixed head (6) are formed, it is characterized in that fixed head (6) is arranged in the box body (5), fixed head (6) is provided with first bottle of hole (6-1), second bottle of hole (6-2), the 3rd bottle of hole (6-3) and the 4th bottle of hole (6-4), immunoglobulin (Ig) reagent bottle (1) is arranged in first bottle of hole (6-1), immune globulin antibody reagent bottle (2) is arranged in second bottle of hole (6-2), phosphate buffer reagent bottle (3) is arranged in the 3rd bottle of hole (6-3), and polyethylene glycol reagents bottle (4) is arranged in the 4th bottle of hole (6-4).

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