JP2000221197A - Kit and method for detecting protein - Google Patents

Abstract

PROBLEM TO BE SOLVED: To make operability superior and simple and to make detection sensitivity satisfactory by forming a detection medium out of polymer fiber and taking a reagent in a reagent solution as a protein error indicator. SOLUTION: A detection medium housing part 8 and a reagent solution housing part 9 are separately prepared by a flexible member 6 on another flexible member 5, to be a support to form spaces for housing a detection medium 1 and a reagent solution, and they are pasted onto the support. A heat seal is peeled off from an opening part 7 to take out the detection medium 1, and the surface of a sample is wiped with the detection medium 1. Then the detection medium 1 is housed in the detection medium housing part 8 again, and the weak heat seals of the detection medium housing part 8 and a reagent solution housing part 9 are peeled away to bring the reagent solution into contact with the detection medium 1 and into reaction with protein on the surface of the detection part of the detection medium 1, without a contact between the reagent solution and a hand. Then it is possible to detect the amount of protein. A protein error indicator among reagents to come into reaction with protein and be discolored is used as the reagent in the reagent solution. Such indicators are widely known as a reagent for detecting protein in urine, and an organic solvent is used partially since it is insoluble in water.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a kit for detecting a protein and a method for detecting a protein.

[0002]

BACKGROUND OF THE INVENTION The present invention relates to a dirt inspection (Hygiene M).
monitoring). The main component of soil in hygiene management, particularly food hygiene management is protein, and a kit that can quickly and easily measure protein for soil has been demanded. JP-A-8-21837 and JP-A-9-5330 propose a method for detecting a protein transferred from a sample surface to a detection medium and a protein detection kit, and are commercially available under the trade name "Fukitori Master". However, there are difficulties in reagent preparation, operability, and detection sensitivity at the time of measurement, and there is a problem in using in a hygiene management site. In addition, the reaction requires time and there is also a problem in the speed.

[0003]

SUMMARY OF THE INVENTION An object of the present invention is to solve the above-mentioned problems, and it is easy to use, has no problem, is quick, and has excellent detection sensitivity when used in a sanitary management site. An object of the present invention is to provide a protein detection kit and a protein detection method.

[0004]

The above object of the present invention is achieved by the following constitution.

[0005] 1. In a protein detection kit comprising a detection medium for transferring a protein from the surface of a sample and a reagent solution that reacts with the protein and discolors, the detection medium is a synthetic polymer fiber, and the reagent in the reagent solution is a protein error indicator. A protein detection kit, comprising:

[0006] 2. A protein detection kit, wherein the protein transferred by the detection medium is brought into contact with the reagent solution on the detection medium, and the amount of protein attached to the surface of the sample is detected based on the degree of discoloration on the detection medium.

[0007] 3. 3. The protein detection kit according to 1 or 2, wherein the detection medium is previously impregnated with water or alcoholic water.

[0008] 4. A protein detection kit, wherein the detection medium and the reagent solution are contained in the same packaging container, and the detection medium and the reagent are brought into contact in the packaging container.

[0009] 5. 5. The protein detection kit according to any one of the above items 1 to 4, wherein the reagent solution comprises one type of solution.

[0010] 6. A protein detection method comprising: bringing a protein transferred to a detection medium into contact with a reagent solution on the detection medium; and detecting the amount of protein attached to the surface of the sample based on the degree of discoloration on the detection medium.

7. 7. The method for detecting a protein according to the above item 6, wherein the reagent solution comprises one type of solution.

Hereinafter, the present invention will be described in more detail.

The present invention will be described below with reference to FIGS. 1 to 3 which are schematic diagrams showing an example of the protein detection kit of the present invention.

The protein detection kit shown in FIG. 1 comprises a container 3 containing a detection medium 1 and an eyedropper container 4 containing a reagent solution 2.

The protein detection kit shown in FIG. 2 is prepared by separating a detection medium storage part 8 and a reagent liquid storage part 9 by another flexible member 6 on a flexible member 5 serving as a support. In addition, a space is created and each is housed and attached to a support. For example, the detection medium 1 and the reagent liquid 2 are separated and stored in a heat-sealed detection medium storage part 8 and a reagent liquid storage part 9. The boundary between the detection medium storage section 8 and the reagent liquid storage section 9 is heat-sealed weaker than the outer edge of the support.

The heat seal is peeled off from the opening 7 in FIG. 2A, the detection medium 1 is taken out, the sample surface is wiped with the detection medium, and then stored again in the detection medium storage section. If you take a weak heat seal between the part and the sample storage part,
The reagent liquid comes into contact with the detection medium, reacts with the protein on the detection medium detection surface 11 without touching the reagent liquid with the hand, and the amount of the protein can be detected.

The container in the package form of the present invention is formed from a plastic film of a flexible member. Specific examples include polyethylene terephthalate (PET), polypropylene (PP), vinylidene chloride, cellulose acetate and the like. Surface treatment that facilitates heat sealing can also be performed.

It is preferable that the boundary between the detection medium storage section and the reagent liquid storage section is weakly heat-sealed. This is because a film whose surface is processed with a material having poor adhesion may be used, Can be achieved by welding at a lower temperature than the surrounding area.

The detection medium of the present invention is a medium capable of transferring proteins on the surface of a sample, for example, a swab-like one comprising a detection part and a shaft, a stick-like detection medium made entirely of the same material, A test paper with a pat is attached.

The material is preferably a hydrophilic synthetic polymer,
For example, a filter obtained by compressing a cotton thread such as acetate, acrylic, polyester, polyurethane or the like, a filter obtained by compressing such a nonwoven fabric, or the like is used.

In the case of a stick, the tip of the detection medium can be obliquely cut so that the protein on the surface can be easily transferred. In addition, the periphery can be wound with a film to prevent contamination due to handling.

The detection medium made of the above material retains the transferred protein and absorbs the remaining reagent solution after the reaction with the reagent, so that the discoloration after the reaction is clear. The detection medium may be pre-moistened with water or water alcohol so that the proteins on the sample surface can be efficiently transferred.

As the reagent in the reagent solution of the present invention, a so-called protein error indicator is used among the reagents which react with proteins and change color. Specifically, for example, tetrabromophenol blue (TBPB), tetrabromophenol phthalene,
5 ', 5 "-dinitro-3', 3" -diiodo-3,
4,5,6-tetrabromophenolsulfophthalane (DINTTB), Coomassie brilliant blue, fast green FCF, light green SF and the like. These protein error indicators are known as reagents for detecting protein in urine. As described in US Pat. No. 4,013,416, an organic solvent is partially used as a solvent for the reagent solution because the reagent is insoluble in water. For example, the content of acetone, ethyl alcohol, methyl cellosolve is 10 to 100%
It is preferably used as an aqueous solution.

The concentration of the reagent in the reagent solution is 0.01% to 0.1%.
% Is preferably used. Since the reagents discolor in different pH ranges, the reagent solution is used in a lower pH range than the discoloration range, usually pH 6 or less, preferably pH 3 or less, but in order to maintain a low pH, hydrochloric acid, acetic acid, citric acid, etc. An acidic agent may be included in the solvent.

The method of using the specific example of the protein detection kit shown in FIG. 1 will be described below.

1. One detection medium 1 is pulled out.

2. The surface of the sample (about 5 cm square) is wiped with the surface of the detection medium detecting section of the detection medium 1. At this time, if the sample surface is dry, it is preferable to wet the sample surface or the detection medium detection unit surface 11 with water or alcohol.

3. Detection medium detection unit surface 11 of detection medium 1
One drop of the reagent solution 2 in the eyedropper container 4 is dropped on the top.

4. The density of the detection medium detection unit surface 11 can be measured by a surface densitometer that compares the density of the detection medium detection unit surface 11 with the color sample 12 (shown in FIG. 3).

[0030]

EXAMPLES The present invention will be described below in detail with reference to examples, but the embodiments of the present invention are not limited thereto.

Embodiment 1 1. Preparation of reagent solution Tetrabromophenol blue (manufactured by Wako Pure Chemical Industries) 10mg
Was dissolved in 50 ml of ethanol, and 1 ml of glacial acetic acid was added to prepare a yellow reagent solution. This reagent solution was placed in an eyedropper container.

2. Preparation of Detection Medium A nonwoven fabric made of acetate fiber was compressed at 150 ° C. while heating at 150 ° C. to form a rod having a length of 12 cm and an outer diameter of 8 mm. Further, the outer periphery was wound with a white PET film having a thickness of 20 μm and heated and welded. The tip was cut at an angle of 60 degrees to produce a detection medium having a length of 9 cm.

3. Preparation of protein sample A 1 mg / ml aqueous solution of bovine serum albumin was prepared and 100 μg / 100 c
It was applied and dried at a ratio of m ² to obtain a protein sample.

4. Detection and Measurement Results After 2 drops of distilled water were dropped on the surface of the protein sample prepared above, an area of 2 × 2 cm was wiped off with the sample medium of the present invention. One drop of the above-prepared reagent solution was dropped on the surface of the sample medium from an eyedropper container. The detection medium that did not wipe the surface used as a control was stained pale yellow, but the surface of the detection medium that was wiped turned blue in part. That is,
The presence of the protein was detectable by the discoloration of the surface of the detection medium detecting portion.

Embodiment 2 1. A reagent solution, a detection medium and a protein sample were prepared in the same manner as in Example 1.

2. Preparation and assembly of packaging container A 100 μm thick polypropylene film was used to form a protein detection kit as shown in FIG.
0 μl is dispensed into the reagent solution storage section 9 shown in FIG.
After installing one of the above-mentioned detection media previously absorbed with 10 ml of 10% ethanol, a 100 μm-thick polypropylene film was heated and welded at 250 ° C. to prepare a protein detection kit shown in FIG. 2 which is an example of the present invention. However, the boundary portion between the sample medium storage part and the reagent liquid storage part was weakly welded at 180 ° C.

3. Comparative Control Kit “Fukitori Master” (purchased from Gunze Sangyo Co., Ltd.) manufactured by Konica Corporation was used as a comparative control kit.

4. Detection Measurement Results The detection medium was taken out of the packaging container, and the protein sample was wiped off 2 × 2 cm and 10 × 10 cm areas. Thereafter, the detection medium was returned to the detection medium storage section shown in FIG. 2, the reagent liquid storage section was pressed, the reagent liquid was extruded, and the surface of the detection medium was brought into contact with the reagent liquid. Immediately, part of the surface turned from light yellow to blue.

Further, the surface density (613 nm) of the discolored reagent medium was measured by a reflection densitometer.

On the other hand, in accordance with the instruction manual for "Fukitori Master", the surface of the protein sample prepared above was wiped off using the wiping solution contained in the kit, and the area of the protein sample surface was measured to be 2 × 2 cm and 10 × 10 cm. Was wiped off. After standing at room temperature for 10 minutes according to the instruction manual, the coloring of the solution was judged by a color scale. Further, the absorbance (562 nm) of the solution was measured by a spectrometer. The results are shown in Table 1 below.

[0041]

[Table 1]

Based on the above results, the protein detection kit and the protein detection method of the present invention have a shorter reaction time than the comparative protein detection kit “Fukitori Master”,
And it turned out that it has high detection sensitivity and has the operability which can be used in the hygiene management spot.

[0043]

EFFECTS OF THE INVENTION The protein detection kit and the protein detection method according to the present invention have good operability and no problem when used in a sanitary control field, and have an effect of rapid detection and excellent detection sensitivity.

[Brief description of the drawings]

FIG. 1 is a schematic diagram showing an example of the protein detection kit of the present invention.

FIG. 2 is a schematic diagram showing an example of the protein detection kit of the present invention.

FIG. 3 is a schematic diagram showing an example of the protein detection kit of the present invention.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Detection medium 2 Reagent liquid 3 Container 4 Eyedropper container 5 Flexible member 6 Flexible member 7 Opening 8 Detection medium storage part 9 Reagent liquid storage part 10 Heat seal 11 Detection medium detection part surface 12 Color sample

Claims (7)

[Claims] 1. A protein detection kit comprising a detection medium for transferring a protein from the surface of a sample and a reagent solution that reacts with the protein and changes color, wherein the detection medium is a synthetic polymer fiber, and the reagent in the reagent solution is Is a protein error indicator. 2. The method according to claim 1, wherein the protein transferred by the detection medium is brought into contact with the reagent solution on the detection medium, and the amount of the protein attached to the surface of the sample is detected based on the degree of discoloration on the detection medium. Protein detection kit. 3. The protein detection kit according to claim 1, wherein the detection medium is previously impregnated with water or alcoholic water. 4. A protein detection kit, wherein the detection medium and the reagent solution are contained in the same packaging container, and the detection medium and the reagent are brought into contact in the packaging container. 5. The protein detection kit according to claim 1, wherein the reagent solution comprises one type of solution. 6. A method for producing a protein, comprising: bringing the protein transferred to the detection medium into contact with a reagent solution on the detection medium; and detecting the amount of protein attached to the surface of the sample based on the degree of discoloration on the detection medium. Detection method. 7. The method for detecting a protein according to claim 6, wherein the reagent solution comprises one type of solution.