CN114807298A - Helicobacter pylori detect reagent box - Google Patents
Helicobacter pylori detect reagent box Download PDFInfo
- Publication number
- CN114807298A CN114807298A CN202210603207.2A CN202210603207A CN114807298A CN 114807298 A CN114807298 A CN 114807298A CN 202210603207 A CN202210603207 A CN 202210603207A CN 114807298 A CN114807298 A CN 114807298A
- Authority
- CN
- China
- Prior art keywords
- helicobacter pylori
- detection kit
- sensitizer
- organic acid
- pylori detection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/58—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving urea or urease
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2304/00—Chemical means of detecting microorganisms
- C12Q2304/40—Detection of gases
- C12Q2304/46—Carbon dioxide
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Toxicology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a helicobacter pylori detection kit; belongs to the field of biotechnology; the detection kit comprises: comprises 13 A container of a C-labelled urea reagent; a container containing a sensitizer; an expiration test bottle; a thin tube for feeding the test bottle; the sensitizer comprises novel organic acid prepared from 6-hydroxy benzopyran-3-formaldehyde and acetic anhydride. The helicobacter pylori detection kit prepared by the invention has higher detection accuracy and precision, and can reduce the content of the helicobacter pylori in the detection kit 13 Amount of C-labeled urea reagent used and 13 abundance of C.
Description
Technical Field
The invention belongs to the technical field of medical instruments, and particularly relates to a helicobacter pylori detection kit.
Background
Helicobacter pylori (Hp) is an important pathogenic bacterium colonized on the surface of gastric mucosa, and is closely related to diseases such as chronic gastritis, peptic ulcer, gastric malignant tumor and the like. Helicobacter pylori mainly spreads through mouth-to-mouth and feces-to-mouth, and is often infected in a family group; thus, early confirmation of H.pylori infection is beneficial for early diagnosis, treatment and prevention of diseases. As helicobacter pylori is fixedly planted on the surface of the epithelial cells of the gastric mucosa, the epithelial cells of the normal gastric mucosa are renewed for 1-3 days, and the helicobacter pylori is discharged along with the fallen gastric mucosa through excrement. The method has the advantages of simple operation, easy acquisition of sample sources and good development prospect in detection of helicobacter pylori. At the same time, use 13 C breath test to diagnose H.pylori infection is also a relatively common diagnostic method. Urea [ 2 ] 13 C]The cost of the reagent is high 13 C one of the main reasons for the high cost of the breath test; urea [ alpha ], [ alpha ] and [ alpha ], [ alpha ] is, and [ alpha ], [ alpha ] is clinically 13 C]In the reagent, the reaction solution is added with a solvent, 13 c abundance is high (generally 99%), and the dosage is large (generally 75 mg), which results in higher cost of the medicament, thereby affecting 13 The extensive use of the C breath test. In addition, the misjudgment rate of patients with upper gastrointestinal hemorrhage and patients who use antacids and anti-Hp drugs recently is higher. How to improve the accuracy rate of the part of patients is the key to further improve the overall accuracy.
Prior art for example publication No. CN101231284A discloses a urea kit for diagnosing helicobacter pylori infection; it specifically relates to a Urea kit for diagnosing helicobacter pylori (Hp) infection by using 13C-UBT (Urea Breath Test), which comprises certain dosage forms of Urea 13 C]Reagents, catheters for insufflation, CO 2 The collection bag also comprises a sensitizer. The kit comprises a sensitizer, reduces the misjudgment rate and ensures that the small dose of urea is used 13 C]The reagent is used or lower 13 Urea of C abundance 13 C]The reagent can also ensure the detection result.
Disclosure of Invention
Objects of the inventionAims to provide a helicobacter pylori detection kit with higher detection accuracy and accuracy, and can reduce the content in the detection kit 13 Amount of C-labeled urea reagent used and 13 abundance of C.
The technical scheme adopted by the invention for realizing the purpose is as follows:
a helicobacter pylori detection kit comprising: comprises 13 A container of a C-labelled urea reagent; a container containing a sensitizer; an expiration test bottle; a thin tube for exhaling into the test bottle;
the sensitizer comprises a novel organic acid;
the novel organic acid has the structural formula
。
The invention adopts novel organic acid as a component of a sensitizer and the sensitizer 13 Co-use of C-labelled urea reagents with reduced content 13 Amount of C-labeled urea reagent used and 13 c abundance, and meanwhile, the detection accuracy and accuracy of the helicobacter pylori detection kit are improved, and the requirements of clinical tests can be met; in addition, the helicobacter pylori detection kit obtained by the specific proportion of the components in the sensitizer has more excellent detection accuracy and accuracy, and has better detection accuracy and accuracy when used for detecting patients with anti-interference drugs such as antacids, anti-Hp drugs and the like.
Further, according to an embodiment of the present invention, in the detection kit 13 The amount of the C-labeled urea reagent is 10-35 mg.
Further, according to an embodiment of the present invention, 13 in C-labelled Urea reagent 13 The abundance ratio of C is 30-60%.
Further, according to an embodiment of the present invention, the amount of the sensitizer is 1.5 to 4.5 g.
Further, according to an embodiment of the present invention, the sensitizer further includes at least one of citric acid, tartaric acid, and ascorbic acid.
Still further, according to an embodiment of the present invention, the sensitizer includes a novel organic acid and citric acid.
Furthermore, according to an embodiment of the present invention, the weight ratio of the novel organic acid to the citric acid in the sensitizer is 0.25-0.55: 1.
further, according to an embodiment of the present invention, the container containing the sensitizer further includes at least one of a sweetener, a preservative, a colorant, and a perfume.
Further, according to an embodiment of the present invention, the novel organic acid is prepared using 6-hydroxybenzopyran-3-carbaldehyde and acetic anhydride.
Further, according to an embodiment of the present invention, a method for preparing a novel organic acid includes: placing anhydrous sodium acetate, acetic anhydride and 6-hydroxy benzopyran-3-formaldehyde into a container, heating and reacting by using an oil pot under the action of stirring, performing rotary evaporation, adding deionized water, filtering, washing, recrystallizing, adding a sodium hydroxide solution for hydrolysis, adjusting the pH of a system to 5-6 by using an acid, performing suction filtration, washing, and recrystallizing to obtain the novel organic acid.
The invention adopts novel organic acid as the component of the sensitizer and the composition containing the sensitizer 13 Co-use of C-labelled urea reagents with reduced content 13 Amount of C-labeled urea reagent used and 13 c abundance, and meanwhile, the detection accuracy and accuracy of the helicobacter pylori detection kit are improved, and the requirements of clinical tests can be met; in addition, the helicobacter pylori detection kit obtained by the specific proportion of the components in the sensitizer has better detection accuracy and accuracy, and the detection accuracy and accuracy of patients using anti-interference medicines such as antacids and anti-Hp medicines are still better. Therefore, the invention is the helicobacter pylori detection kit with higher detection accuracy and accuracy, and simultaneously can reduce the content in the detection kit 13 Amount of C-labeled urea reagent used and 13 abundance of C.
Drawings
FIG. 1 is an infrared spectrum of the novel organic acid of example 1.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, and the examples are given only for illustrating the present invention and not for limiting the scope of the present invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
According to a preferred embodiment of the present invention, a helicobacter pylori detection kit comprises:
1) a is composed of 13 A container of a C-labelled urea reagent; wherein 13 The amount of the C-labeled urea reagent is 10-35 mg, and 13 the abundance of C is 30-60%;
2) a container containing a sensitizer, wherein the sensitizer is prepared into a sensitizer aqueous solution;
3) designating a test bottle containing the patient's breath;
4) a thin tube for exhaling into the test bottle.
Another object of the present invention is to provide a method for using a helicobacter pylori detection kit, which comprises the steps of:
(a) preparing sensitizer into sensitizer aqueous solution, and taking one dose 13 C-labelled Urea reagent, Observation of pathology, urease and 13 c, the result of the breath test;
(b) a thin tube for exhaling into the test bottle is adopted, and the test bottle is used for collecting the exhaled gas of the patient; firstly, long expiration is carried out, the gas exhaled by the front part is not collected, and only the gas exhaled by the rear part is collected.
Further, according to an embodiment of the present invention, the method for preparing the sensitizer aqueous solution comprises:
and (3) completely dissolving the sensitizer, sodium carbonate, spice and sweetener in deionized water, and uniformly stirring and mixing to obtain a sensitizer aqueous solution with the pH value of 2.2-2.8.
Furthermore, according to an embodiment of the present invention, the sensitizer is 1.5-4.5 g, the sodium carbonate is 0.075-0.15 g, the perfume is 0.025-0.045 g, the sweetener is 0.035-0.065 g, and the deionized water is 100-200 g.
Further, according to an embodiment of the present invention, in the method for preparing the novel organic acid, 2.0 to 3.5 parts by weight of anhydrous sodium acetate, 8 to 15 parts by weight of acetic anhydride, 2.5 to 4.5 parts by weight of 6-hydroxybenzopyran-3-carbaldehyde, 50 to 100 parts by weight of deionized water, and 20 to 40 parts by weight of a sodium hydroxide solution are added.
Further, according to an embodiment of the present invention, in the method for preparing a novel organic acid, the concentration of the sodium hydroxide solution is 5 to 10 wt%.
Further, according to an embodiment of the present invention, in the method for preparing the novel organic acid, the heating reaction temperature is 125 to 150 ℃, and the reaction time is 4 to 8 hours.
The technical solution of the present invention is further described in detail below with reference to the following detailed description and the accompanying drawings:
example 1:
the preparation method of the novel organic acid comprises the following steps:
placing 2.5 parts by weight of anhydrous sodium acetate, 10 parts by weight of acetic anhydride and 3 parts by weight of 6-hydroxychroman-3-formaldehyde into a three-neck flask, heating to 135 ℃ by using an oil pot under the stirring action, reacting for 6 hours, removing a solvent by rotary evaporation, adding 60 parts by weight of deionized water to dissolve the sodium acetate, filtering, washing with deionized water, recrystallizing by using chloroform as a solvent, adding 25 parts by weight of 7.5wt% sodium hydroxide solution for hydrolysis, adjusting the pH of a system to 5.5 by using 5wt% dilute sulfuric acid, performing suction filtration, washing with deionized water, and recrystallizing by using hot water to obtain a novel organic acid, wherein the structural formula of the novel organic acid is shown in the specification
。
Performing nuclear magnetic characterization on the strain by using a nuclear magnetic resonance spectrometer (VNMRS type 600), 1 H NMR（DMSO，600MHz）：11.2（s，1H，COOH）、7.49（s，1H，CH）、6.83（s，1H，CH）、6.78（d，1H，CH）、6.72（d，1H，CH）、6.37（s，1H，CH）、5.38（s，1H，OH）、5.31（d，1H，CH）、3.25（s，2H，CH 2 ）。
example 2:
a helicobacter pylori detection kit comprising:
1) a is composed of 13 A blister container of C-labelled urea reagent; wherein 13 The amount of the C-labeled urea reagent was 12mg, and 13 the abundance of C is 40%;
2) a sachet containing 2.5g of sensitiser wherein the weight ratio of the novel organic acid of example 1 to citric acid in the sensitiser is 0.25: 1, preparing a sensitizer into a sensitizer aqueous solution;
3) designating a test bottle containing the patient's breath;
4) a thin tube for exhaling into the test bottle.
In this embodiment, the preparation method of the sensitizer aqueous solution comprises:
2.5g of sensitizer, 0.1g of sodium carbonate, 0.03g of lemon flavor (purchased from Jiangsu Caesami Biotech Co., Ltd.), and 0.045g of aspartame were completely dissolved in 150g of deionized water, and were stirred and mixed uniformly to obtain a sensitizer aqueous solution having a pH of 2.5.
Example 3:
a helicobacter pylori detection kit, which is different from that of example 2, and which comprises: 1) one comprises 13 A blister container of C-labelled urea reagent; wherein 13 The amount of the C-labeled urea reagent was 20mg, and 13 the abundance of C was 30%.
Example 4:
a helicobacter pylori detection kit, which is different from that of example 2, and which comprises: 2) a sachet containing 4.5g of sensitiser wherein the weight ratio of the novel organic acid of example 1 to citric acid in the sensitiser is 0.25: 1, preparing the sensitizer into a sensitizer aqueous solution.
In this embodiment, the preparation method of the sensitizer aqueous solution is as follows:
4.5g of sensitizer, 0.1g of sodium carbonate, 0.03g of lemon flavor (purchased from Jiangsu Caesami Biotech Co., Ltd.), and 0.045g of aspartame were completely dissolved in 150g of deionized water, and were stirred and mixed uniformly to obtain a sensitizer aqueous solution having a pH of 2.5.
Example 5:
a helicobacter pylori detection kit, which is different from that of example 2, and which comprises: 2) a sachet containing 2.5g of sensitiser wherein the weight ratio of the novel organic acid of example 1 to citric acid in the sensitiser is 0.55: 1, preparing the sensitizer into a sensitizer aqueous solution.
In this example, the method of preparing the sensitizer aqueous solution was the same as in example 2.
Example 6:
a helicobacter pylori detection kit, which is different from that of example 2, and which comprises: 2) a sachet containing 2.5g of sensitiser comprising only the novel organic acid of example 1, the sensitiser being formulated as an aqueous sensitiser solution.
In this example, the method of preparing the sensitizer aqueous solution was the same as in example 2.
Example 7:
a helicobacter pylori detection kit, which is different from that of example 2, and which comprises: 2) a sachet containing 2.5g of sensitiser, wherein the sensitiser comprises only citric acid, the sensitiser being formulated as an aqueous sensitiser solution.
In this example, the method of preparing the sensitizer aqueous solution was the same as in example 2.
In order to further improve the detection accuracy and sensitivity of the detection kit, 0.015-0.035 g of 11-keto-beta-boswellic acid is added in the preparation process of the sensitizer aqueous solution; which interact with other components of the aqueous sensitizer solution and contain them in small doses 13 C-labelled urea reagent or 13 The urea reagent with low C abundance can also obtain an accurate detection result.
Example 8:
a helicobacter pylori detection kit, which is different from that of example 2, and which comprises: 2) a sachet containing 2.5g of sensitiser wherein the weight ratio of the novel organic acid of example 1 to citric acid in the sensitiser is 0.25: 1, preparing a sensitizer into a sensitizer aqueous solution;
in this embodiment, the preparation method of the sensitizer aqueous solution is as follows:
2.5g of sensitizer, 0.1g of sodium carbonate, 0.03g of lemon spice (purchased from Jiangsu Caesami Biotechnology Co., Ltd.), 0.02g of 11-keto-beta-boswellic acid and 0.045g of aspartame are placed in 150g of deionized water to be completely dissolved, and are stirred and mixed uniformly to obtain a sensitizer aqueous solution with the pH value of 2.5.
Example 9:
a helicobacter pylori detection kit, which is different from the embodiment 6, in the embodiment, the preparation method of the sensitizer aqueous solution comprises the following steps:
2.5g of sensitizer, 0.1g of sodium carbonate, 0.03g of lemon spice (purchased from Jiangsu Caesami Biotechnology Co., Ltd.), 0.02g of 11-keto-beta-boswellic acid and 0.045g of aspartame are placed in 150g of deionized water to be completely dissolved, and are stirred and mixed uniformly to obtain a sensitizer aqueous solution with the pH value of 2.5.
Example 10:
a helicobacter pylori detection kit, which is different from the embodiment 7, in the embodiment, the preparation method of the sensitizer aqueous solution comprises the following steps:
2.5g of sensitizer, 0.1g of sodium carbonate, 0.03g of lemon spice (purchased from Jiangsu Caesami Biotechnology Co., Ltd.), 0.02g of 11-keto-beta-boswellic acid and 0.045g of aspartame are placed in 150g of deionized water to be completely dissolved, and are stirred and mixed uniformly to obtain a sensitizer aqueous solution with the pH value of 2.5.
Example 11:
a method for using a helicobacter pylori detection kit comprises the following steps:
(a) the helicobacter pylori detection kit in example 2 was used; preparing the sensitizer into sensitizer aqueous solution, and taking a dose 13 C-labelled Urea reagent, Observation of pathology, urease and 13 c, the result of the breath test;
(b) a thin tube for exhaling into the test bottle is adopted, and the test bottle is used for collecting the exhaled gas of the patient; firstly, long expiration is carried out, the gas exhaled by the front part is not collected, and only the gas exhaled by the rear part is collected.
Example 12:
a method of using a helicobacter pylori detection kit, which is different from that of example 11: in the step (a), the helicobacter pylori detection kit of example 2 was replaced with the helicobacter pylori detection kit of example 3.
Example 13:
a method of using a helicobacter pylori detection kit, which is different from that of example 11: in the step (a), the helicobacter pylori detection kit of example 2 was replaced with the helicobacter pylori detection kit of example 4.
Example 14:
a method of using a helicobacter pylori detection kit, which is different from that of example 11: in the step (a), the helicobacter pylori detection kit of example 2 was replaced with the helicobacter pylori detection kit of example 5.
Example 15:
a method of using a helicobacter pylori detection kit, which is different from that of example 11: in the step (a), the helicobacter pylori detection kit of example 2 was replaced with the helicobacter pylori detection kit of example 6.
Example 16:
a method of using a helicobacter pylori detection kit, which is different from that in example 11: in the step (a), the helicobacter pylori detection kit of example 2 was replaced with the helicobacter pylori detection kit of example 7.
Example 17:
a method of using a helicobacter pylori detection kit, which is different from that of example 11: in the step (a), the helicobacter pylori detection kit of example 2 was replaced with the helicobacter pylori detection kit of example 8.
Example 18:
a method of using a helicobacter pylori detection kit, which is different from that of example 11: in the step (a), the helicobacter pylori detection kit of example 2 was replaced with the helicobacter pylori detection kit of example 9.
Example 19:
a method of using a helicobacter pylori detection kit, which is different from that of example 11: in the step (a), the helicobacter pylori detection kit of example 2 was replaced with the helicobacter pylori detection kit of example 10.
Comparative example 1:
a helicobacter pylori detection kit comprising:
1) a is composed of 13 A blister container of C-labelled urea reagent; wherein 13 The amount of C-labeled urea reagent is 50mg, and 13 the abundance of C is 99%;
2) designating a test bottle containing the patient's breath;
3) a thin tube for exhaling into the test bottle.
Comparative example 2:
a method of using a helicobacter pylori detection kit, which is different from that of example 11: in step (a), the helicobacter pylori detection kit of example 2 was replaced with the helicobacter pylori detection kit of comparative example 1.
Test example:
1. infrared structural characterization
The novel organic acid prepared in example 1 was characterized by infrared spectroscopy using a Fourier infrared spectrometer (Nicolet 67 model) with a scanning range of 500-4000cm -1 。
FIG. 1 is a graph of the infrared spectrum of the novel organic acid of example 1; as can be seen from FIG. 1, at 3400cm -1 The characteristic absorption peak appearing nearby is the stretching vibration of the phenolic hydroxyl; at 1700cm -1 The characteristic absorption peak appearing nearby is the stretching vibration of C = O in carboxyl; at 1625cm -1 、1515cm -1 Stretching vibration with a characteristic absorption peak of C = C occurring in the vicinity; at 1050cm -1 The characteristic absorption peak appearing nearby is the stretching vibration of C-O-C; thus, the novel organic acid is prepared by adopting 6-hydroxy benzopyran-3-formaldehyde and acetic anhydride.
2. Test result of helicobacter pylori detection kit
(1) Subject: 50 helicobacter pylori infected patients (no antacid or anti-Hp medicine is used recently) are positive in DOB more than or equal to 2.4 per mill and negative in DOB less than 2.4 per mill; firstly adopting the commercial standard 13 Helicobacter pylori detection kit for C breath test (i.e., C breath test) 13 Amount of C-labeled Urea reagentIs 75mg, and 13 abundance of C99%), followed by detection according to the detection methods in examples 11-19, statistical analysis of the data using SPSS 21.0 software; the diagnosis accuracy is evaluated by using a Kappa index, and the consistency is good when the consistency is more than 0.75; 0.75-0.04 is better in consistency; less than 0.4 is poor in consistency; and drawing an ROC curve to analyze the diagnostic value of the helicobacter pylori detection kit; lower accuracy at AUC of 0.5-0.7; 0.7-0.9 has certain accuracy; higher accuracy is achieved when the accuracy is higher than 0.9.
TABLE 1 13 Test results of C breath test
As can be seen from Table 1, in examples 11 to 14, the Kappa index is not less than 0.8, and the AUC is higher than 0.9, which indicates that the detection result of the helicobacter pylori detection kit used has good consistency with the commercially available standard helicobacter pylori detection kit, and the detection accuracy is high; comparing example 11 with comparative example 2, the Kappa index and AUC in example 11 are higher than those in comparative example 2, and those in the H.pylori detection kit used in example 11 13 The amount of C-labeled urea reagent was lower than that of comparative example 2, and 13 the abundance of C is also much lower than in comparative example 2, indicating that the sensitizers and compositions used in the present invention 13 Co-use of C-labelled urea reagents with reduced content 13 Amount of C-labeled urea reagent used and 13 the abundance of C improves the detection accuracy and accuracy of the helicobacter pylori detection kit.
In addition, comparing example 11 with examples 15 to 16, the Kappa index and AUC in example 11 are higher than those in examples 15 to 16, which shows that the novel organic acid prepared by using 6-hydroxybenzopyran-3-carbaldehyde and acetic anhydride according to the present invention as a sensitizer component, and the sensitizer component is mixed with a mixture containing the same 13 The urea reagent marked by C is used together, so that the detection accuracy of the helicobacter pylori detection kit can be improved, the diagnosis accuracy is higher, and the requirements of clinical tests can be met.
As can be seen from Table 1, the Kappa index in example 17 is higher than 0.93, AUC is higher than 0.95, the Kappa index and AUC in comparative example 11 and example 17, example 15 and example 18, example 16 and example 19, both Kappa index and AUC in example 17 are higher than those in example 11, both Kappa index and AUC in example 18 are higher than those in example 15, and both Kappa index and AUC in example 19 are higher than those in example 16, which indicates that the addition of 11-keto-beta-boswellic acid to the aqueous sensitizer solution further improves the detection accuracy and precision of the helicobacter pylori detection kit.
(2) Influence of sensitizer component on helicobacter pylori detection kit
For the helicobacter pylori detection kit in example 2, the weight ratio of the novel organic acid to citric acid in the sensitizer component was changed, that is, the weight ratio of the novel organic acid to citric acid in example 1 in 2.5g of the sensitizer was 0.15: 1. 0.25: 1. 0.35: 1. 0.45: 1. 0.55: 1. 0.65: 1 and divided into test group a1, test group a2, test group A3, test group a4, test group a5, and test group a 6; wherein the usage method of the helicobacter pylori detection kit is the same as that of the embodiment 11, and the detection method is the same as that of the commercially available standard 13 Helicobacter pylori detection kit for C breath test (i.e., C breath test) 13 The amount of the C-labeled urea reagent was 75mg, and 13 the abundance of C was 99%) and the analysis was performed in the same manner as in (1).
TABLE 2 13 Test results of C breath test
As can be seen from Table 2, the Kappa index in the test group A2-test group A5 is higher than 0.82, the AUC is higher than 0.91, and is higher than the test group A1 and the test group A6, which indicates that the weight ratio of the novel organic acid to the citric acid in the sensitizer added in the helicobacter pylori detection kit is 0.25-0.55: 1, the obtained helicobacter pylori detection kit has more excellent detection accuracy and accuracy.
(3) Subject: 50 H.pylori-infected patients recently administered antacids and anti-Hp drugs; other tests and analysis methods were the same as in (1).
TABLE 3 13 Test results of C breath test
As can be seen from Table 3, when patients infected with helicobacter pylori using an antacid and an anti-Hp drug are detected, the Kappa index is higher than 0.7 and the AUC is higher than 0.83 in examples 11 to 14, which indicates that the detection result of the helicobacter pylori detection kit prepared by the invention has better consistency with the commercially available standard helicobacter pylori detection kit, higher detection accuracy and certain detection accuracy, i.e., the detection accuracy and accuracy of the helicobacter pylori detection kit are not greatly influenced by the use of the drug; comparing example 11 with comparative example 2, the Kappa index and AUC in example 11 are higher than those in comparative example 2, and those in the H.pylori detection kit used in example 11 13 The amount of C-labelled urea reagent is lower than in comparative example 2, and 13 the abundance of C is also much lower than in comparative example 2, indicating that the sensitizers and compositions used in the present invention 13 Co-use of C-labelled urea reagents with reduced content 13 Amount of C-labeled urea reagent used and 13 abundance of C, i.e. using small doses containing 13 The C-labeled urea reagent has better detection accuracy and accuracy for patients who use the medicine. Comparing example 11 with examples 15-16, wherein the Kappa index and AUC in example 11 are higher than those in examples 15-16, it is demonstrated that the novel organic acid prepared by using 6-hydroxybenzopyran-3-carbaldehyde and acetic anhydride as the sensitizer component of the present invention, and the sensitizer component is mixed with the organic acid containing 13 The urea reagent marked by C is used together, so that the detection accuracy of the helicobacter pylori detection kit can be improved, the diagnosis accuracy is higher, the misjudgment rate is reduced, and the requirement of clinical tests can be better met.
As can be seen from Table 3, the Kappa index in example 17 is higher than 0.82, the AUC is higher than 0.93, the Kappa index and AUC in comparative example 11 and example 17, example 15 and example 18, example 16 and example 19, both the Kappa index and AUC in example 17 are higher than those in example 11, both the Kappa index and AUC in example 18 are higher than those in example 15, and both the Kappa index and AUC in example 19 are higher than those in example 16, which indicates that the addition of 11-keto-beta-boswellic acid to the aqueous sensitizer solution further improves the detection accuracy and precision of the helicobacter pylori detection kit.
Conventional operations in the operation steps of the present invention are well known to those skilled in the art and will not be described herein.
The embodiments described above are intended to illustrate the technical solutions of the present invention in detail, and it should be understood that the above-mentioned embodiments are only specific embodiments of the present invention, and are not intended to limit the present invention, and any modification, supplement or similar substitution made within the scope of the principles of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A helicobacter pylori detection kit, comprising: comprises 13 A container of a C-labelled urea reagent; a container containing a sensitizer; an expiration test bottle; a thin tube for feeding the test bottle;
the sensitizer comprises a novel organic acid;
the novel organic acid has the structure of
。
2. The helicobacter pylori detection kit according to claim 1, wherein: in the detection kit 13 The amount of the C-labeled urea reagent is 10-35 mg.
3. The helicobacter pylori detection kit according to claim 1, wherein: the above-mentioned 13 In C-labelled Urea reagent 13 The abundance of C is 30-60%.
4. The helicobacter pylori detection kit according to claim 1, wherein: the amount of the sensitizer is 1.5-4.5 g.
5. The helicobacter pylori detection kit according to claim 1, wherein: the sensitizer also comprises at least one of citric acid, tartaric acid and ascorbic acid.
6. The helicobacter pylori detection kit according to claim 5, wherein: the sensitizer comprises novel organic acid and citric acid.
7. The helicobacter pylori detection kit according to claim 6, wherein: the weight ratio of the novel organic acid to the citric acid in the sensitizer is 0.25-0.55: 1.
8. the helicobacter pylori detection kit according to claim 1, wherein: the container containing the sensitizer also comprises at least one of a sweetener, a preservative, a coloring agent and a spice.
9. The helicobacter pylori detection kit according to claim 1, wherein: the novel organic acid is prepared from 6-hydroxy benzopyran-3-formaldehyde and acetic anhydride.
10. The helicobacter pylori detection kit according to claim 9, wherein: the preparation method of the novel organic acid comprises the following steps: placing anhydrous sodium acetate, acetic anhydride and 6-hydroxy benzopyran-3-formaldehyde into a container, heating and reacting by using an oil pot under the action of stirring, performing rotary evaporation, adding deionized water, filtering, washing, recrystallizing, adding sodium hydroxide for hydrolysis, adjusting the pH of a system to 5-6 by using acid, performing suction filtration, washing, and recrystallizing to obtain the novel organic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210603207.2A CN114807298B (en) | 2022-05-31 | 2022-05-31 | Helicobacter pylori detect reagent box |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210603207.2A CN114807298B (en) | 2022-05-31 | 2022-05-31 | Helicobacter pylori detect reagent box |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114807298A true CN114807298A (en) | 2022-07-29 |
| CN114807298B CN114807298B (en) | 2022-10-25 |
Family
ID=82519518
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210603207.2A Active CN114807298B (en) | 2022-05-31 | 2022-05-31 | Helicobacter pylori detect reagent box |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114807298B (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20010064675A (en) * | 2000-05-31 | 2001-07-11 | 강성훈 | Low-enriched 13C-Labelled Urea Composition for Urea Breath Test |
| US20030032081A1 (en) * | 2001-08-09 | 2003-02-13 | Sitke Aygen | Method for the diagnosis of Helicobacter pylori infection, and a diagnostic kit for performing the method |
| CN101231284A (en) * | 2007-01-23 | 2008-07-30 | 张厚德 | Urea kit for diagnosing pylorus helicobacterium infestation |
| CN101887062A (en) * | 2009-05-15 | 2010-11-17 | 艾克拜尔·热合曼 | Helicobacter pylori detection kit containing 13C-urea solution |
| CN103757088A (en) * | 2014-01-20 | 2014-04-30 | 四川万可泰生物技术有限责任公司 | Quantitative determination and drug allergy determination kits for helicobacter pylori viable bacteria and determination method |
| CN111088312A (en) * | 2019-12-26 | 2020-05-01 | 欧迪玛(湖北)生物科技有限公司 | Helicobacter pylori detection kit and preparation method thereof |
| CN112695066A (en) * | 2021-01-29 | 2021-04-23 | 郑州安图生物工程股份有限公司 | Drug sensitivity kit for culturing and identifying helicobacter pylori and detection method |
| CN113621681A (en) * | 2020-05-08 | 2021-11-09 | 修正生物医药（杭州）研究院有限公司 | Composition and kit for detecting helicobacter pylori in oral cavity |
-
2022
- 2022-05-31 CN CN202210603207.2A patent/CN114807298B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20010064675A (en) * | 2000-05-31 | 2001-07-11 | 강성훈 | Low-enriched 13C-Labelled Urea Composition for Urea Breath Test |
| WO2001092889A1 (en) * | 2000-05-31 | 2001-12-06 | Medichems Co., Inc. | Low-enriched 13c-labelled urea composition for urea breath test |
| US20030032081A1 (en) * | 2001-08-09 | 2003-02-13 | Sitke Aygen | Method for the diagnosis of Helicobacter pylori infection, and a diagnostic kit for performing the method |
| CN101231284A (en) * | 2007-01-23 | 2008-07-30 | 张厚德 | Urea kit for diagnosing pylorus helicobacterium infestation |
| CN101887062A (en) * | 2009-05-15 | 2010-11-17 | 艾克拜尔·热合曼 | Helicobacter pylori detection kit containing 13C-urea solution |
| CN103757088A (en) * | 2014-01-20 | 2014-04-30 | 四川万可泰生物技术有限责任公司 | Quantitative determination and drug allergy determination kits for helicobacter pylori viable bacteria and determination method |
| CN111088312A (en) * | 2019-12-26 | 2020-05-01 | 欧迪玛(湖北)生物科技有限公司 | Helicobacter pylori detection kit and preparation method thereof |
| CN113621681A (en) * | 2020-05-08 | 2021-11-09 | 修正生物医药（杭州）研究院有限公司 | Composition and kit for detecting helicobacter pylori in oral cavity |
| CN112695066A (en) * | 2021-01-29 | 2021-04-23 | 郑州安图生物工程股份有限公司 | Drug sensitivity kit for culturing and identifying helicobacter pylori and detection method |
Non-Patent Citations (2)
| Title |
|---|
| KARAM CHAND ET AL.: "Synthesis,1Hand13C NMR assignment of novel2-pyridone derivatives", 《MAGN RESON CHEM.》 * |
| 刘娅琳 等: "幽门螺杆菌细菌培养与尿素酶试验对幽门螺杆菌检测敏感性比较", 《贵阳医学院学报》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114807298B (en) | 2022-10-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1101546C (en) | Method and kit for detecting helicobacter pylori | |
| CN101977633A (en) | Process for preparing a pharmaceutical formulation of contrast agents | |
| CN101031324A (en) | Diagnostic agent for tumor | |
| CN103251962A (en) | Magnetic resonance contrast material and preparation method thereof, and contrast agent | |
| CN114807298B (en) | Helicobacter pylori detect reagent box | |
| WO2010128787A9 (en) | Gadolinium complex, method for preparing same, and mri contrast agent comprising same | |
| CN1249070C (en) | Paramagnetic metal-phthalocyanine complex compounds and contrast agent using the same | |
| CN103623435A (en) | Nanometer magnetic resonance contrast material, nanometer magnetic resonance contrast agent and preparation method of nanometer magnetic resonance contrast agent | |
| CN110927156A (en) | Urine detection kit for cervical cancer and preparation method thereof | |
| CN110357828B (en) | Nuclear magnetic resonance contrast agent and preparation method and application thereof | |
| US4351823A (en) | Diagnosis of tumors or bacterial infections having β-glucuronidase activity | |
| CN100509840C (en) | Tubeimuside and bile acid derivative and their medicine composition | |
| CN102716076A (en) | Ambroxol hydrochloride medicine combination for injection | |
| CN115266663B (en) | Biological probe for non-invasive diagnosis of parkinsonism triggered by intestinal microenvironment as well as preparation method and application thereof | |
| CN101698105A (en) | Gastrointestinal tract contrast medium | |
| CN115461052B (en) | Pharmaceutical use of complexes of ARB metabolites with NEP inhibitors for the prevention and/or treatment of kidney disease | |
| US11673909B2 (en) | Furaneone glycoside compound, pharmaceutical composition thereof, preparation method therefor, and application thereof | |
| JPH01100187A (en) | Metal-containing saccharides-polysulfate compound and manufacture, diagnostic agent for detecting gastrointestinal diseases, manufacture and nmr- and x ray diagnostic agent | |
| CN106589022B (en) | A kind of roxithromycin compound and preparation method thereof, pharmaceutical composition | |
| AU2001256768B2 (en) | Preparations for diagnosing infection with helicobacter pylori | |
| CN114053222A (en) | Digoxin injection and preparation method thereof | |
| CN101881763A (en) | Urea breath detection kit for detecting stomach helicobacter pylori | |
| CN110105380A (en) | A kind of muscle PET imaging molecule probe and the preparation method and application thereof | |
| WO2023062386A1 (en) | Method for the synthesis of evoc probes | |
| CN109602920A (en) | A kind of high stability novel dendritic molecular image probe and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |