CN101019764A - Pyloric helicobacter detecting method - Google Patents
Pyloric helicobacter detecting method Download PDFInfo
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- CN101019764A CN101019764A CN 200710013360 CN200710013360A CN101019764A CN 101019764 A CN101019764 A CN 101019764A CN 200710013360 CN200710013360 CN 200710013360 CN 200710013360 A CN200710013360 A CN 200710013360A CN 101019764 A CN101019764 A CN 101019764A
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- urease
- helicobacter pylori
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Abstract
The pyloric helicobacter detecting method has conventional conductivity meter or resistance meter used to detect the change of the measured posture in resistance value or conductivity so as to determine the urease value and detect pyloric helicobacter. The method includes the steps of: preparing pyloric helicobacter detecting chip and urease detecting solution, inserting the detecting chip and a gastroscope into the stomach to be detected, detecting the change of conductivity based on the detecting chip sensed parameters, converting into the urease value, and determining the content of pyloric helicobacter. The present invention has reasonable detection scheme, high detection accuracy and high safety, and may be used widely in fast detection of pyloric helicobacter.
Description
The technical field is as follows:
the invention relates to a helicobacter pylori detection method, in particular to a method for detecting the existence of helicobacter pylori by preparing a detection chip and preparing a detected solution and measuring the urease content by utilizing the change of the conductivity or the resistance value. The method is suitable for measuring the section tissue of the gastric mucosa and is also suitable for being inserted into the stomach to be matched with a gastroscope to measure the helicobacter pylori on the stomach wall.
Background art:
helicobacter Pylori (HP) has been known as a bacterial source responsible for gastrointestinal disorders including gastric and duodenal ulcers since its discovery in 1983 by Warren and Marshall, WHO has identified it as a typical carcinogen. The methods for diagnosing helicobacter pylori can be classified into two major categories, namely, non-invasive examination and invasive examination. Non-invasive tests include serological methods, PCR on gastric juice and breath tests with isotopic labels. The traumatic examination is completed by means of gastroscopy, including morphology, microbiology and the like. The serological method is to extract blood to be tested for helicobacter pylori antibody. The specific antibody will not appear after several weeks of helicobacter pylori infection, and cross-reactive antibodies also exist in the blood of the person without helicobacter pylori infection, and the antibodies in the blood can be maintained at positive level for a long time (more than 6 months) after the helicobacter pylori is eradicated. Serological methods are therefore also unsuitable as a clinical routine diagnostic method.
The gastric juice PCR method is to suck gastric juice through a nasogastric tube and extract DNA for PCRamplification to diagnose helicobacter pylori infection, and has the advantages of good accuracy and high sensitivity, but the operation is complex, and false positive is easily caused by pollution, so the method cannot be listed as routine inspection. The microbiological examination method is the most accurate method for diagnosing helicobacter pylori infection, but certain anaerobic culture conditions are required besides gastroscopy biopsy, time is consumed, technical requirements are high, popularization is not easy, and false negative is easy to occur due to the relation of biopsy parts. The morphological method requires taking a living tissue, smearing, staining a section and observing by a microscope. Although false negative is likely to occur depending on the relationship between the site of live tissue, it is still a method generally used clinically. Commercial kits currently tested using urease are "CLOtest" (USPNo.4748113), "Hpfast" (USPNo.5439801), and "Pyloritek" (USPNo.5314804 and 5420016). There is another patent (CN 24588626) which utilizes a modified ion sensitive field effect transistor. However, the method in the prior art generally has the outstanding defects of complex testing procedure, inconvenient use and operation, low detection accuracy, non-visual detection result, low safety factor and the like.
The invention content is as follows:
the invention aims to overcome the defects of the prior art and find a novel helicobacter pylori detection method which is suitable for measuring the section tissues of gastric mucosa and is also suitable for being inserted into the stomach to be matched with a gastroscope to directly measure the helicobacter pylori infection condition of the stomach wall.
In order to achieve the purpose, the urease value is measured by measuring the resistance value or the change of the conductivity of the detected body position by using a conventional conductivity meter or a conventional resistance meter, so that the effect of detecting the helicobacter pylori is achieved; preparing a helicobacter pylori detection chip and a urease detection solution, inserting the detection chip and a gastroscope into the stomach of a detected body in a matching manner, determining the change value of the conductivity by using a resistance meter or a conductivity meter according to the sensing parameters of the detection chip, then converting and determining the existence value of urease, and then determining the content of the helicobacter pylori according to the relationship between the urease and the helicobacter pylori and the chemical decomposition reaction process of urea.
The helicobacter pylori measured by the method is rich in urease, urea generates carbonic acid and ammonia water under the action of the urease, the carbonic acid and the ammonia water are further ionized, electric conductivity or resistance of a conductive part is changed by electric ions, and the change value reflects the content of the urease, so that the content of the helicobacter pylori is reflected, and the detection purpose is realized. The total reaction formula of the urea decomposition reaction process is as follows:
the stepwise reaction equation is as follows:
The above reaction process involved in the present invention shows that, as the reaction proceeds, the number of positive and negative ions in the solution increases, thereby changing the original conductivity or resistance value, and that the conductivity or resistance value of the conductive part is measured by a conventional resistance or conductivity measuring instrument device, so that urease can be rapidly detected and the presence of helicobacter pylori can be detected.
The detection chip used by the invention is made into a cuboid by taking a non-conductive PE high molecular plate material as a substrate, two long sides on the front surface of the detection chip are respectively printed with parallel and alternate ink electrode layers, an insulating layer with a circular hole with the diameter of 2-4mm dug in the center is covered on the electrode layer, an induction zone containing urea is covered on the circular hole of the insulating layer, the detection chip is electrically communicated with a conductivity or resistance measuring device to form a detection system of helicobacter pylori, and the detection chip is directly inserted into the stomach to be matched with a gastroscope, so that the helicobacter pylori in the gastric mucosa can be directly measured.
The preparation method of the urease measured solution comprises the steps of weighing a certain amount of urease freeze-dried powder, dissolving the urease freeze-dried powder in distilled water, weighing a certain amount of the solution, placing the solution in a urea solution with the concentration of 0.2g/ml, mixing, and preparing the urease measured solution.
Compared with the prior art, the method has the advantages of reasonable design of the detection scheme, high detection accuracy, good safety, convenience in use and operation, visual numerical value reflection and wide application in the rapid detection of the helicobacter pylori of gastrointestinal patients.
Description of the drawings:
FIG. 1 is a schematic diagram of the structural principle of a detection chip prepared and used in the present invention, which includes a PE polymer sheet 1; a conductive printing ink electrode layer 2; an insulating layer 3 with pores with the diameter of 2-4 mm; sensing zone 4.
FIG. 2 is a schematic diagram of the present invention for measuring the conductivity change of urease solutions with different concentrations.
FIG. 3 is a schematic diagram of the present invention for measuring the resistance change of urease in slice tissue by using a detection chip.
The specific implementation mode is as follows:
the invention is further illustrated by the following examples. The method determines the urease value by determining the resistance value or the change of the conductivity of the detected body position by means of a conventional conductivity meter or a resistance meter, thereby achieving the effect of detecting the helicobacter pylori; preparing a helicobacter pylori detection chip and a urease detection solution, inserting the detection chip and a gastroscope into the stomach of a detected body in a matching manner, measuring the change value of the conductivity by using a resistance meter or a conductivity meter according to the sensing parameters of the detection chip, then converting and measuringthe existence value of urease, and determining the content of the helicobacter pylori according to the relationship between the urease and the helicobacter pylori and the chemical decomposition reaction process of urea. The detection chip comprises a PE polymer sheet 1, a conductive printing ink electrode layer 2, an insulating layer 3 and a sensing zone 4, wherein the non-conductive PE polymer sheet material is used as a substrate to manufacture a cuboid sheet 1, the two long side sides of the front surface of the cuboid sheet 1 are respectively printed with the parallel and alternate printing ink electrode layers 2, the insulating layer 3 with a round hole with the diameter of 2-4mm dug in the center is covered on the electrode layer 2, the round hole of the insulating layer 3 is covered with the sensing zone layer 4 containing urea, the detection chip is electrically communicated with a conductivity or resistance measuring device to form a detection system of helicobacter pylori, the detection chip is directly inserted into the stomach to be matched with a gastroscope, so that the helicobacter pylori of gastric mucosa is directly measured, or slice tissues are placed.
Example 1:
preparation of urease test solution: firstly weighing 5g of urease freeze-dried powder, dissolving the urease freeze-dried powder in 10.00mL of distilled water, weighing 1mL of urease solution, and inversely putting the urease solution in 24mL of urea solution with the concentration of 0.2g/mL for mixing to prepare a urease solution to be detected; inserting an electric conduction electrode into the solution to be detected and electrically communicating the electric conduction electrode with a conductivity meter by using a lead; mixing the urease solution and the urea solution to be used as a timing zero point, and measuring the change of the conductivity within ten minutes; observing and recording the conductance in urease-containing solutions containing different concentrationsThe rate was varied with time as shown in FIG. 2, wherein a, b, c, and d are urease concentrations of 3.2X 10-4，2.4×10-4，1.6×10-4，0.8×10-4Change in conductivity at g/mL.
Example 2:
manufacturing a detection chip: cutting a polycarbonate plate with the thickness of 0.2 mm into cuboid small pieces with the length of 10 mm and the width of 10 mm, respectively printing two parallel ink electrode layers on two long side sides of the front surface of the polycarbonate plate by utilizing conductive carbon paste in a printing mode, covering an insulating layer with a small hole with the diameter of 3mm at the center of the upper surface of each electrode layer, partially covering the parallel electrodes, dripping urea solution in an induction zone of the insulating layer, and airing; placing the gastric mucosa tissue slice in a sensing area of the detection chip, and slightly squeezing; electrically connecting the two exposed parts of the two parallel electrodes with a resistivity or resistance measuring instrument by using a lead; the change of the electrical conductivity was measured within 10 minutes by placing the gastric mucosa tissue section in the sensing area of the detection chip as a timing zero point, as shown in FIG. 3, wherein a, b, c, d are the resistance change at different urease concentrations.
Claims (4)
1. A method for detecting pyloric helicobacterium includes such steps as preparing pyloric helicobacterium detecting chip and urease solution, inserting said chip in stomach of object, measuring the variation of conductivity by said chip, converting to determine the value of urease, and determining the content of pyloric helicobacterium according to the relation between urease and pyloric helicobacterium and the chemical decomposition reaction of urea.
2. The method according to claim 1, wherein the helicobacter pylori is determined to be rich in urease, urea is produced into carbonic acid and ammonia water under the action of urease, the carbonic acid and ammonia water are further ionized, and the electric conductivity or resistance of the electric conducting part is changed by the ionization, and the change value reflects the content of urease, so that the content of helicobacter pylori is reflected.
3. The method for detecting helicobacter pylori according to claim 1, wherein the detection chip is a cuboid made of a non-conductive PE polymer plate material as a substrate, and ink electrode layers are printed on two long sides of the front surface of the detection chip in parallel and alternately, an insulating layer with a circular hole with a diameter of 2-4mm dug in the center is covered on the electrode layer, a sensing region containing urea is covered on the circular hole of the insulating layer, and the detection chip is electrically communicated with a conductivity or resistance measuring device to form a detection system for helicobacter pylori.
4. The method according to claim 1, wherein the urease test solution is prepared by dissolving urease lyophilized powder in distilled water, and mixing the above solutions in 0.2g/ml urea solution to obtain urease test solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200710013360 CN101019764A (en) | 2007-03-05 | 2007-03-05 | Pyloric helicobacter detecting method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200710013360 CN101019764A (en) | 2007-03-05 | 2007-03-05 | Pyloric helicobacter detecting method |
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| CN101019764A true CN101019764A (en) | 2007-08-22 |
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| CN 200710013360 Pending CN101019764A (en) | 2007-03-05 | 2007-03-05 | Pyloric helicobacter detecting method |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102316786A (en) * | 2009-02-17 | 2012-01-11 | 西门子公司 | Endoscopic capsule |
| CN104880489A (en) * | 2015-06-25 | 2015-09-02 | 合肥美的暖通设备有限公司 | Method for testing rinsing performance of alkaline degreaser |
| CN106834412A (en) * | 2015-12-03 | 2017-06-13 | 李和楼 | A kind of method of inspection of helicobacter pylori |
| CN107037117A (en) * | 2017-06-02 | 2017-08-11 | 华东医院 | The Rapid identification and Virulent Analysis method of a kind of helicobacter pylori |
| CN108085279A (en) * | 2017-12-29 | 2018-05-29 | 重庆大学 | In a kind of calcareous sand urease-producing bacterium efficiently separate and identification method |
| CN113406153A (en) * | 2021-06-29 | 2021-09-17 | 毕玉琦 | Food flora short-term test device |
| CN114107531A (en) * | 2021-12-17 | 2022-03-01 | 石家庄市疾病预防控制中心（石家庄市卫生检测中心） | Detection and application method of stomach micro-ecological urease positive flora |
-
2007
- 2007-03-05 CN CN 200710013360 patent/CN101019764A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102316786A (en) * | 2009-02-17 | 2012-01-11 | 西门子公司 | Endoscopic capsule |
| US8918154B2 (en) | 2009-02-17 | 2014-12-23 | Siemens Aktiengesellschaft | Endoscopic capsule |
| CN104880489A (en) * | 2015-06-25 | 2015-09-02 | 合肥美的暖通设备有限公司 | Method for testing rinsing performance of alkaline degreaser |
| CN106834412A (en) * | 2015-12-03 | 2017-06-13 | 李和楼 | A kind of method of inspection of helicobacter pylori |
| CN107037117A (en) * | 2017-06-02 | 2017-08-11 | 华东医院 | The Rapid identification and Virulent Analysis method of a kind of helicobacter pylori |
| CN108085279A (en) * | 2017-12-29 | 2018-05-29 | 重庆大学 | In a kind of calcareous sand urease-producing bacterium efficiently separate and identification method |
| CN113406153A (en) * | 2021-06-29 | 2021-09-17 | 毕玉琦 | Food flora short-term test device |
| CN114107531A (en) * | 2021-12-17 | 2022-03-01 | 石家庄市疾病预防控制中心（石家庄市卫生检测中心） | Detection and application method of stomach micro-ecological urease positive flora |
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