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Beta-glucuronidase detection reagent, reaction pad, preparation method of reaction pad and kit

A technology of β-glucuronidase and detection reagent, which is applied in its preparation method and kit, β-glucuronidase detection reagent, and reaction pad field, can solve the problems such as the lack of accuracy of β-glucuronidase, and achieve High sensitivity, accurate detection, simple and convenient use conditions

Inactive Publication Date: 2017-05-10
GUANGZHOU HONGQI OPTICAL INSTR TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection reagents currently used to detect β-glucuronidase are not accurate enough

Method used

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  • Beta-glucuronidase detection reagent, reaction pad, preparation method of reaction pad and kit
  • Beta-glucuronidase detection reagent, reaction pad, preparation method of reaction pad and kit
  • Beta-glucuronidase detection reagent, reaction pad, preparation method of reaction pad and kit

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Experimental program
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preparation example Construction

[0051] The preparation method of this reaction pad comprises the following steps:

[0052] 1) Material preparation: prepare polyvinylpyrrolidone-K90, 5-bromo-4-chloro-3-indole-β-D-glucuronide and neonatal bovine serum according to the above parts by weight;

[0053] 2) Prepare phosphate buffered saline (PBS): prepare phosphate buffered saline with a pH of 6.8 to 7.5 for use;

[0054] The phosphate buffer is NaH 2 PO 4and Na 2 HPO 4 aqueous solution;

[0055] 3) Preparation of soaking solution A: dissolving polyvinylpyrrolidone and 5-bromo-4-chloro-3-indole-β-D-glucuronide sodium in phosphate buffer solution to obtain soaking solution A;

[0056] 4) Prepare soaking solution B: fully dissolve newborn bovine serum in phosphate buffer solution to obtain soaking solution B;

[0057] 5) Soaking solution A treatment: put the filter paper in the soaking solution A and fully soak it for 10-20 seconds, then take it out, and then dry it at a temperature of 20-70°C for 30-40 minutes...

Embodiment 1～3

[0062] Embodiments 1 to 3 disclose β-glucuronidase detection reagents, respectively prepare three groups of raw material components according to Table 1, and prepare different sodium phosphate buffer solutions with different concentrations and pHs according to Table 2; Group A is used in Embodiment 1, B Group C is used in Example 2, and Group C is used in Example 3.

[0063] Dissolve polyvinylpyrrolidone, 5-bromo-4-chloro-3-indole-β-D-glucuronide sodium, and neonatal bovine serum in sequence in 100 mL of sodium phosphate buffer to obtain β-glucuronidase assay reagent.

[0064] Table 1 The composition and content of the detection reagent

[0065]

[0066]

[0067] The preparation parameter of table 2 sodium phosphate buffer solution

[0068]

[0069] Detect the detection reagents obtained in Examples 1 to 3: prepare respectively β-glucuronidase standard substance solutions with different enzyme activity concentrations, β-glucuronidase standard substance solutions use...

Embodiment 4～6

[0077] Embodiment 4~6 discloses a kind of reaction pad and preparation method thereof:

[0078] 1) Prepare materials according to Table 1, prepare sodium phosphate buffer solution according to Table 2; Group A is used in Example 4, Group B is used in Example 5, and Group C is used in Example 6;

[0079] 2) Preparation of soaking solution A: dissolving polyvinylpyrrolidone and sodium 5-bromo-4-chloro-3-indole-β-D-glucuronide in 100 mL of sodium phosphate buffer to obtain soaking solution A;

[0080] 3) Preparation of soaking solution B: Fully dissolve newborn bovine serum in 100 mL of sodium phosphate buffer to obtain soaking solution B;

[0081] 4) Soaking solution A treatment: put the filter paper in the soaking solution A and fully soak it for 10-20 seconds, then take it out, and then dry it at a temperature of 40°C for 30-40 minutes to obtain the soaking solution A filter paper;

[0082] 5) Soaking solution B treatment: soak the filter paper of soaking solution A in soakin...

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Abstract

The invention discloses a beta-glucuronidase detection reagent, a reaction pad, a preparation method of the reaction pad and a kit. The beta-glucuronidase detection reagent is prepared from polyvinylpyrrolidone, 5-bromo-4-chloro-3-indole-beta-D-glucuronide and new-born calf serum. The preparation method of the reaction pad comprises steps as follows: 1), polyvinylpyrrolidone and 5-bromo-4-chloro-3-indole-beta-D-glucuronide are dissolved in a phosphate buffer solution, and a soak solution A is obtained; 2), the new-born calf serum is dissolved in the phosphate buffer solution, and a soak solution B is obtained; 3), a carrier is put in the soak solution A and is sufficiently soaked and dried, and a soak solution A carrier is obtained; 4), the soak solution A carrier is put in the soak solution B and is sufficiently soaked and dried, and the reaction pad is obtained. According to the detection reagent, the stability of activity of beta-glucuronidase is specifically enhanced through addition of the new-born calf serum, so that the detection is more accurate.

Description

technical field [0001] The invention relates to the field of aerobic vaginitis detection reagents, in particular to a β-glucuronidase detection reagent, a reaction pad, a preparation method and a kit. Background technique [0002] Vaginitis is an inflammation of the vaginal mucosa and submucosal connective tissue, and is a common disease in gynecological clinics. For normal healthy women, due to anatomical and biochemical characteristics, the vagina has a natural defense function against the invasion of pathogens. When the natural defense function of the vagina is destroyed, pathogens are easy to invade and cause vaginal inflammation. There are three types of common vaginitis: bacterial vaginosis (BV / AV), fungal vaginitis (VVC) and trichomonas vaginitis (TV). Among them, female bacterial vaginal infection can be divided into two types, one is caused by anaerobic bacteria and facultative anaerobic bacteria, clinically called bacterial vaginosis (BV), and the other is caused ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573
CPCG01N33/573
Inventor 眭红燕朱华琳范静彦李必松
Owner GUANGZHOU HONGQI OPTICAL INSTR TECH