CN111289505A

CN111289505A - Helicobacter pylori detect reagent box

Info

Publication number: CN111289505A
Application number: CN202010184232.2A
Authority: CN
Inventors: 张广明
Original assignee: Nanjing Kangrong Health Technology Co Ltd
Current assignee: Nanjing Kangrong Health Technology Co Ltd
Priority date: 2020-03-16
Filing date: 2020-03-16
Publication date: 2020-06-16

Abstract

The application provides a kit capable of rapidly detecting helicobacter pylori, the kit adopts a rapid culture method to shorten the detection time, and the kit also uses ammonia time-receiving agent and buffer pair to increase the detection accuracy and reduce the misjudgment rate of clinical medical staff. The kit can be used for large-scale clinical screening to prevent helicobacter pylori infection and transmission.

Description

Helicobacter pylori detect reagent box

Technical Field

The application relates to the field of medical equipment, in particular to a kit for detecting helicobacter pylori, and belongs to the field of medicines.

Background

Helicobacter pylori lives in the pylorus and oral cavity of the human stomach, and is one of the most common bacterial pathogens. Most of the world's population is infected with H.pylori, and in some countries almost 90% of people are infected with this bacterium. People are usually infected at early age, reaching 50% under 5 years of age. This bacterial infection first causes chronic gastritis and leads to gastric ulcers and gastric atrophy, and in severe cases gastric cancer.

According to statistics, the incidence rate of atrophic gastritis and gastric cancer of people with early age of the initial helicobacter pylori infection is high, and the helicobacter pylori infection and the death rate of gastric cancer are in parallel. Helicobacter pylori is parasitic in the tissues of the gastric mucosa, and 67% to 80% of gastric ulcers and 95% of duodenal ulcers are caused by helicobacter pylori. The common symptoms of patients with chronic gastritis and peptic ulcer are: after eating, the upper abdomen is full, uncomfortable or painful, often accompanied by other adverse symptoms such as belching, abdominal distension, acid regurgitation, anorexia, etc. Some patients may also have recurrent severe abdominal pain, minor upper gastrointestinal bleeding, etc. Therefore, experts think that people who find helicobacter pylori infection as soon as possible can kill helicobacter pylori with antibiotics timely and effectively, and the method has great significance for preventing and controlling gastric cancer.

Currently, there are many methods for detecting helicobacter pylori infection, such as biopsy, isolated culture of helicobacter pylori, rapid urease test, urea breath test, urinary ammonia excretion test, serology test, polymerase chain reaction, and the like. The method adopted by different hospitals is notSimilarly, most hospital procedures are specific, rapid, and some are non-invasive. The principle of the method is as follows: (1) antibody antigen method; (2) a bacterial culture method; (3) urease method. The antibody anti-method is used because it has a detection defect for a history of secondary infection with H.pylori and a double diagnosis after H.pylori treatment, and is now used only for rough detection of H.pylori. The urease method is characterized in that helicobacter pylori can secrete high-activity urease, and the urease decomposes urea to generate CO₂And NH₃. The urea breath test is carried out by detecting and marking CO₂And fast urease is NH determination₃. Currently on the market by detecting NH₃The accuracy of (1) NH is generally low₃The amount of the catalyst is low; (2) NH (NH)₃Is easy to volatilize. So NH₃The length of the stay time on the reagent box test paper is a key for improving the detection accuracy. CN1266461 discloses a urease kit for endoscope, which is composed of urea and KH₂PO₄Hypochlorite, phenate solution and phenol red. The main action principle is that urease decomposes urea to produce CO₂And NH₃Then, the indigo method is used for detecting trace ammonia to judge the helicobacter pylori infection, and in the system, gel made of agar is used as a carrier of the whole system. Because hypochlorite and phenate have poor stability and the carrier is agar gel, urea is decomposed in advance in the system, so that the substrate urea is insufficient during detection, and accurate detection cannot be performed. And the detection result needs 1 to 3 hours.

Helicobacter pylori produces urease, and the urease needs a certain time to decompose urea, so that the conventional rapid urease method needs more time for detecting each sample, thereby being not beneficial to large-scale screening development. The application adopts a super-rapid culture method to ensure that helicobacter pylori is nourished, so as to rapidly generate urease and accelerate the generation of substances for identification by decomposing a substrate urea. The application also solves a problem which puzzles the accuracy of the method: this application can absorb the ammonia absorbent of ammonia through adding on the kit, fixes ammonia on the kit, can strengthen the color reaction like this, makes the phenomenon more obvious, has more to plant in medical personnel's judgement. The present application also addresses the accuracy problem from another aspect: the indicator used on the kit needs to change color within a certain pH value range, the pH value of saliva or gastric juice is not necessarily just in the color change range, for example, phenol red is used as the indicator, the color change range is 6.8-8.2, the indicator is yellow when the pH value is lower than 6.8, the indicator is changed from yellow to red when the pH value is higher than 6.8, the indicator is red when the pH value is higher than 8.2, and the indicator can be changed into red when the pH value is higher than 7.2. If saliva is used as a sample, the pH of the saliva itself may be greater than 6.8, which may discolor the kit, and the kit may be judged to be positive. The kit prepared by the application uses trace buffer salt, the color change point of the kit can be corrected in time, if helicobacter pylori infection exists, the ammonia generated by urea decomposition by urease can break the balance, and the whole reaction system is changed to the direction larger than 6.8. The amount of the buffer salt is strictly controlled when the kit is prepared, the purpose of not using is achieved when the amount is small, and the formed balance system is not easy to break when the amount is large, so that the color is not changed.

Disclosure of Invention

The application aims to provide a helicobacter pylori detection kit capable of rapidly detecting helicobacter pylori, and the kit can inform medical staff of helicobacter pylori infection of patients in a short time.

The helicobacter pylori detection kit is characterized by comprising urea, an alkaline gas absorbent, a pH indicator, a culture medium and pH value buffer salt.

The helicobacter pylori detection kit is characterized in that the alkaline gas absorbent is one or more of citric acid and monovalent salt, tartaric acid and salt, boric acid, nickel chloride, aluminum sulfate, aluminum trichloride and copper sulfate, wherein the nickel chloride, aluminum sulfate and aluminum trichloride are preferred.

The helicobacter pylori detection kit is characterized in that the culture medium is one or more of 199 culture medium, BME culture medium, 1640 culture medium, RPMI-1640 culture medium, DMEM-F12 culture medium, DMEM culture medium, F10 culture medium, F12 culture medium, Fischer's culture medium, IMDM culture medium and MEM culture medium.

The helicobacter pylori detection kit is characterized in that the pH value buffer salt is one or more of acetic acid/sodium acetate, ammonia/ammonium chloride, phosphoric acid/sodium phosphate or potassium phosphate, disodium hydrogen phosphate/sodium dihydrogen phosphate, phthalic acid buffer salt and citric acid buffer salt.

The helicobacter pylori detection kit is characterized in that the alkaline gas absorbent for capturing alkaline gas and the buffer salt with the adjusted pH value exist on the test paper in the kit in a solid form, and can be quickly dissolved when a small amount of liquid exists.

The helicobacter pylori detection kit is characterized in that the other form of the kit is that an alkaline gas absorbent for capturing alkaline gas, a buffer salt with a pH value adjusted back are prepared into a reagent A, a culture medium and a substrate urea are used as a solid reagent B, and a pH indicator is fixed on a test paper.

The helicobacter pylori detection kit is characterized in that when the helicobacter pylori infection is detected, the kit only needs to stick a cotton swab to a small amount of saliva, then the cotton swab stuck with the saliva is used for wiping the inner side of teeth to sample, the sampled cotton swab is coated on a reaction test paper of the helicobacter pylori detection kit, and the reaction test paper is compared with a colorimetric card within 1-3 minutes, so that whether the helicobacter pylori infection exists is judged.

The helicobacter pylori detection kit as described above is characterized in that the kit is characterized in that the reagent B is added to the reagent A when detecting whether helicobacter pylori is infected, and the reagent B is completely dissolved to obtain a mixed reagent solution. And (3) coating a small amount of saliva or gastric mucosa tissue on a reaction test paper of the helicobacter pylori detection kit, dropwise adding the mixed reagent solution, and comparing the reaction test paper with a colorimetric card for 8 to 10 minutes so as to judge whether the helicobacter pylori infection exists.

Detailed Description

Example 1

Respectively dissolving 3 parts of urea, 1 part of nickel chloride, 0.7 part of 1640 culture medium and 0.5 part of sodium acetate in 100 parts of purified water, dissolving 0.1 part of phenol red in proper ethanol, adding the dissolved phenol red into the purified water, stirring to uniformly mix, adjusting the pH value to 6.5-6.7 by using acetic acid, soaking filter paper in the prepared solution for 12 hours, taking out and airing, cutting the filter paper into strips, and packaging the strips in a kit.

Example 2

2.5 parts of urea, 0.8 part of aluminum sulfate, 0.7 part of DMEM-F12 culture medium and 0.5 part of sodium phosphate are respectively dissolved in 100 parts of purified water, 0.1 part of bromothymol blue is dissolved in proper ethanol, the purified water is added, the mixture is stirred and uniformly mixed, the pH value is adjusted to be 5.7-5.9 by acetic acid, filter paper is soaked in the prepared solution for 12 hours, the filter paper is taken out, aired, cut into strips and packaged in a kit.

Example 3

0.8 part of aluminum trichloride and 0.5 part of citric acid buffer salt in the prescription amount are respectively dissolved in 100 parts of purified water to prepare a reagent A, and the pH value is adjusted to 3.5-3.7 by hydrochloric acid. Urea powder 2.5 parts and DMEM medium powder 0.7 parts were thoroughly mixed to prepare granules as reagent B. Dissolving 0.2 parts of bromocresol green indicator in 50 parts of purified water, soaking filter paper in the prepared solution for 12 hours, taking out, airing, cutting into strips, and packaging in the kit.

Test example 1

89 clinical patients with primary diagnosis of gastritis or chronic gastritis are selected, the kits prepared in the embodiment 1 are respectively used for testing, at least three mucosal tissue samples are respectively taken for culturing in a mode of gastroscope biopsy mucosal tissue culture after testing, and the helicobacter pylori infection condition is detected. The results show that the kit detects 82 positive patients, and the positive rate is 93.26%. The positive rate of 84 patients tested positive by the gastroscope biopsy mucosa culture was 94.38%. The two detection methods are positive, and the coincidence rate of the two detection methods is 94.05 percent, wherein the number of the patients is 79.

The foregoing is a more detailed description of the present application in connection with specific embodiments thereof, and it is not intended that the present application be limited to the specific embodiments thereof. For those skilled in the art to which the present application pertains, several simple deductions or substitutions may be made without departing from the spirit of the present application, which should be considered as belonging to the protection scope of the present application.

Claims

1. A helicobacter pylori detection kit for detecting helicobacter pylori comprises urea, an alkaline gas absorbent, a pH indicator, a culture medium and pH buffer salt.

2. The helicobacter pylori detection kit according to claim 1, wherein the alkaline gas absorbent is one or more of citric acid and monovalent salt, tartaric acid and salt, boric acid, nickel chloride, aluminum sulfate, aluminum trichloride and copper sulfate, and preferably nickel chloride, aluminum sulfate and aluminum trichloride.

3. The helicobacter pylori detection kit according to claim 1, wherein the medium is one or more of 199 medium, BME medium, 1640 medium, RPMI-1640 medium, DMEM-F12 medium, DMEM medium, F10 medium, F12 medium, Fischer's medium, IMDM medium, and MEM medium.

4. The helicobacter pylori detection kit according to claim 1, wherein the pH buffer salt is one or more selected from the group consisting of acetic acid/sodium acetate, ammonia/ammonium chloride, phosphoric acid/sodium phosphate or potassium phosphate, disodium hydrogen phosphate/sodium dihydrogen phosphate, phthalic acid buffer salt, and citric acid buffer salt.

5. A helicobacter pylori detection kit according to claim 1, wherein the kit is in the form of an alkaline gas absorbent for capturing alkaline gas, a pH-adjusted buffer salt, a culture medium, a substrate urea and a pH indicator, which are present in a solid form on a test paper in the kit and are rapidly reconstituted in the presence of a small amount of liquid.

6. The helicobacter pylori detection kit according to claim 1, wherein the kit is further characterized in that an alkaline gas absorbent for capturing alkaline gas, a buffer salt preparation reagent A with a pH adjusted back, a culture medium and a substrate urea are used as a solid reagent B, and a pH indicator is fixed on the test paper.

7. The helicobacter pylori detection kit according to claim 5, wherein the kit is characterized in that only a small amount of saliva is adhered to a cotton swab when detecting whether the helicobacter pylori is infected, then the cotton swab adhered with the saliva is used for wiping the inner side of a tooth to sample, the sampled cotton swab is coated on a reaction test paper of the helicobacter pylori detection kit, and the reaction test paper is compared with a colorimetric card for 1 to 3 minutes so as to judge whether the helicobacter pylori is infected.

8. The helicobacter pylori detection kit according to claim 6, wherein the kit is characterized in that the reagent B is added to the reagent A to dissolve the whole reagent A to obtain a mixed reagent solution when detecting whether helicobacter pylori is infected. And (3) coating a small amount of saliva or gastric mucosa tissue on a reaction test paper of the helicobacter pylori detection kit, dropwise adding the mixed reagent solution, and comparing the reaction test paper with a colorimetric card for 8 to 10 minutes so as to judge whether the helicobacter pylori infection exists.