WO2014067032A1

WO2014067032A1 - Method and kit for detecting haemoglobin in rectal exudate

Info

Publication number: WO2014067032A1
Application number: PCT/CN2012/001559
Authority: WO
Inventors: 温鑫; 崔子伦; 杜晓丽
Original assignee: 青岛贝尔奥生物科技有限公司
Priority date: 2012-10-30
Filing date: 2012-11-19
Publication date: 2014-05-08
Also published as: CN102914538A; CN102914538B

Abstract

Provided is a method for detecting haemoglobin in rectal exudate, comprising: wiping at the site of recta under slight pressure to obtain exudate; treating the exudate with a trimethylol aminomethane-hydrochloric acid buffer of PH 7.4; reacting with a chromogenic reagent and performing color comparison with a colorimetric plate; and recording detection results. Also provided is a kit for detecting haemoglobin in rectal exudate.

Description

Method and test box for detecting rectal exudate hemoglobin

[0001] The present invention relates to a detection method, and in particular to a method for detecting hemolysin in rectal exudate, and the present invention also relates to a special detection kit for detecting hemolysin in rectal exudate.

Background technique

[0002] Heme in rectal exudate is often caused by heme release from tissue cell oxygen receptors and FixL transmembrane proteins, hemoglobin precipitation from tumor cells or microcirculation exudation, and its hemoglobin content is extremely small, which is invisible to the naked eye. In view of the fact, it is generally necessary to use advanced instruments and analytical chemistry methods for testing. The cost is high, the cycle is long, and the operation is complicated. Therefore, a simple and low-cost detection method is invented to quickly determine whether hemoglobin is contained in the rectal exudate. A problem that needs to be solved.

Summary of the invention

[0003] In order to solve the problems of high cost, long cycle, and complicated operation in the method for detecting rectal exudate hemoglobin in the prior art, the present invention provides a method for detecting rectal exudate hemoglobin, which has high sensitivity and low cost. The hemoglobin in the rectal exudate can be quickly detected; and the present invention also provides a special detection kit for detecting hemoglobin in the rectal exudate.

[0004] The technical solution of the present invention is as follows:

The method for detecting hemoglobin in rectal exudate provided by the invention comprises the following detection steps:

1) Bring the latex gloves, put the finger cot on the first knuckle of the index finger, and apply a little pressure on the rectum to obtain the exudate; in which, the exudate volume is 0.5~lml;

2) Put the finger sleeve with liquid exudation into the sample cup. The sample cup contains trishydroxymethylaminocarbamidine-hydrochloric acid buffer. The amount of buffer is 3ml, and the mixture is mixed to obtain the sample solution. Preferably, the trishydroxyl The pH of methylaminoformamide-hydrochloric acid buffer was 7.4;

3) Add 1 ml of sample solution, 0.5 ml of reagent A, and 0.5 ml of reagent B in the test cell; mix and observe the color change within 2 minutes, and record the test results with the color plate; the preparation method of the reagent A is as follows: 50 mg of 3,3',5,5'-tetramethylbenzidine was dissolved in 100 ml of a pH 4.4 citrate buffer; the reagent B was a 3% hydrogen peroxide solution.

[0005] The invention also provides a detection box for detecting rectal exudate hemoglobin, comprising a box body and a sliding cover slidingly connected with the box body, wherein the sliding cover is provided with two strip-shaped grooves protruding outward The strip groove is provided with a drop tube, and the strip groove is sealed from the bottom. The inside of the box body is hollow, and one side is provided with a color swatch plate, and a test cup and a sample cup placing area for placing the sample cup are further provided at the bottom of the box body, and the test is performed. A finger sleeve is placed at the edge of the pool, and the test pool extends inward from the bottom of the box body, and the middle portion is recessed downward into a bowl-like structure.

[0006] Further, the sample cup placement area is disposed between the color palette and the test cell. [0007] Further, the color palette is parallel to the bottom of the box body and located at the upper middle portion of the box body.

[00081 Further, the dropper is provided with a scale.

[0009] Further, the sample cup is provided with a sealing cover, and the cup of the sample cup has a cylindrical shape.

[0010] Further, the material of the finger sleeve is an absorbent nonwoven fabric.

[0011] The detection method provided by the invention is simple and practical, low in cost, and can quickly screen samples containing no hemoglobin in the rectal exudate, samples containing heme and samples containing hemoglobin, for further examination. Preparation; The minimum hemoglobin concentration detectable by the invention is up to 0.5 ng/L, and the sensitivity is high; and the invention also provides a detection box, which can be used exclusively for holding the utensils and reagents used in the detection process, the detection The box has a simple structure, and the entire detection step can be completed with only one detection box, which saves space and reagents, and also reduces the labor burden on the operator.

DRAWINGS

1 is a schematic view showing the external structure of the detection box;

Figure 2 is a schematic view of the internal structure of the casing;

Figure 3 is a schematic view of the structure of the finger sleeve;

Figure 4 is a schematic view of the sample cup structure;

Among them: 1. Slide cover; 2. Strip groove; 3. Sample cup placement area; 4. Test cell; 5. Color palette; 6. Finger sleeve; 7. Sample cup; 8. Box body.

detailed description

[0013] The present invention will be further described below in conjunction with the embodiments.

[0014] Embodiment 1

A method for detecting rectal exudate hemoglobin is as follows: Slide the slide cover 1 of the test box, take out the finger sleeve 6 from the box body 8, put on the latex glove, and then wear the finger sleeve 6 at the first joint of the index finger. Upper, in the rectum, a little pressure to wipe the exudate, wherein the amount of exudate is 0.5-1 ml; take the sample cup 7 of the sample cup placement area 3, and put in the trimethylolamine-hydrochloric acid with a pH of 7.4. 3 ml of buffer, put the finger-strap 6 with liquid exudation into the sample cup 7, and mix; take two dropper from the two strips 2, the two drops are pre-loaded with reagent A and reagent B, in the test cell 4, sequentially add 1 ml of sample liquid, 0.5 ml of reagent A, 0.5 ml of reagent B, mix, observe the color change within 2 minutes, and record the detection result with the color plate 5; preparation of the reagent A The method is as follows: 50 mg of 3,3',5,5'-tetramethylbenzidine is dissolved in 100 ml of a pH 4.4 citrate buffer; the reagent B is a 3% hydrogen peroxide solution.

[0015] The test result is determined as follows:

If it does not develop color, it is negative, indicating that there is no hemoglobin in the exudate; if it shows light green blue to light blue, it may contain hemoglobin, which needs to be tested to determine; if it shows blue to dark blue, it indicates that it is in the exudate. Contains heme. [0016] To determine the sensitivity of the detection method provided by the present invention, a sample of different colors is further verified by a mass spectrometer as follows:

Three samples were detected as non-color-developing, showing light green blue to light blue, blue to dark blue, and the heme was detected by mass spectrometer. The test results were as follows: None of the colores were detected. And hemoglobin; light green to light blue: 0.5 ng / L, 1.7 ng / L, 4.3 ng / L; blue to dark blue is 20 ng / L, 2.5 μ g / L, 52 ug / L.

[0017] It can be seen from the above data that the detection method provided by the invention can make the exudate of the hemoglobin content less than 1 ng/L develop color, and the exudate of the hemoglobin content of 20 ng/L or more can be determined to be contained in the exudate. Heme, so the detection method provided by the invention has high sensitivity and can be used for qualitative detection of hemoglobin in rectal exudate.

[0018] Example 2

The following describes the special detection box of the present invention in conjunction with FIG. 1 to FIG.

[0019] The detection box for detecting rectal exudate hemoglobin provided by the invention comprises a box body 8 and a sliding cover 1 slidably connected with the box body 8, and the sliding cover 1 is slidable along the upper surface of the box body 8, the sliding cover 1 is provided with two strip-shaped grooves 2 which are outwardly convex. The strip-shaped groove 2 is provided with a dropper, the strip-shaped groove 2 is sealed from the bottom, the inside of the box 8 is hollow, and one side of the box body 8 is provided with colorimetric The bottom of the box body 8 is further provided with a test cell 4 and a sample cup placement area 3 on which the sample cup 7 is placed. A test sleeve 4 is placed on the side of the test cell 4, and the test cell 4 extends inward from the bottom of the case body 8, The middle portion is recessed downward into a bowl-like structure, and the sample cup placement area 3 is disposed between the color palette 5 and the test cell 4, and the color palette 5 is parallel to the bottom of the case 8 and located at the upper middle portion of the case 8. The dropper tube is provided with a scale, the sample cup 7 is provided with a sealing cover, the cup of the sample cup 7 is cylindrical, and the material of the finger sleeve 6 can be an absorbent non-woven fabric.

Claims

Claim

A method for detecting rectal exudate hemoglobin, comprising the following detecting steps:

1) Put on the latex gloves, put the finger cot on the first knuckle of the index finger, and apply a little pressure on the exudate to obtain the exudate;

2) Put the finger sleeve with liquid exudation into the sample cup. The sample cup contains 0.05mol/L trishydroxymethylaminocarbamidine-hydrochloric acid buffer solution, the amount of buffer is 3ml, and it is mixed to obtain the sample liquid;

3) Add 1 ml of sample solution, 0.5 ml of reagent A, and 0.5 ml of reagent B in the test cell; mix and observe the color change within 2 minutes, and record the test results with the color plate; the preparation method of the reagent A is as follows: 50 mg of 3,3',5,5'-tetramethylbenzidine was dissolved in a pH 4.4 citrate buffer of 100 mr; the reagent B was a 3% hydrogen peroxide solution.

2. Method according to claim 1, characterized in that the amount of exudate is 0.5-1 ml.

The method according to claim 1 or 2, wherein the pH of the tris(hydroxymethyl)carbamidine-hydrochloric acid buffer is 7.4.

4. A detection box for detecting rectal exudate hemoglobin, comprising a box body and a sliding cover slidingly connected with the box body, wherein the sliding cover is provided with two strip-shaped grooves protruding outward, and the strip-shaped groove is filled therein The dropper, the strip groove is sealed from the bottom, the inside of the box body is hollow, and one side is provided with a color swatch plate, and a test cup and a sample cup placing area for placing the sample cup are further arranged at the bottom of the box body, and a finger sleeve is placed on the test pool side. The test cell extends inward from the bottom of the box body, and the middle portion is recessed downward into a bowl-like structure.

The test cartridge according to claim 4, wherein the sample cup placement area is disposed between the color palette and the test cell.

The test cartridge according to claim 4, wherein the colorimetric plate is parallel to the bottom of the case and is located at a middle upper portion of the case.

7. The test cartridge according to claim 4, wherein the dropper is provided with a scale.

The test cartridge according to claim 4, wherein the sample cup is provided with a sealing cover, and the cup of the sample cup has a cylindrical shape.

The test cartridge according to claim 4, wherein the finger cuff is made of an absorbent nonwoven fabric.