Novel biomarker detection method for early gastric cancer screening

Abstract

The invention relates to the technical field of early gastric cancer screening, and discloses a novel biomarker detection method for early gastric cancer screening, which comprises the following steps: S1, preparation before detection: performing joint detection on pepsinogen, gastritin-17, carcino-embryonic antigen and carbohydrate antigen 72-4, and preparing corresponding detection reagents and standard substances; s2, sample collection: adopting a disposable sterile blood taking needle and a vacuum tube, and extracting 3-5ml of venous blood through sterile operation; s3, sample treatment and storage: separating serum through a centrifugal machine, and after the treatment is completed, putting the sample into a storage device for storage; and S4, detection operation: preheating the detector, and then calibrating the detector by a standard product. In the invention, the pepsinogen, the gastritin-17, the carcino-embryonic antigen and the carbohydrate antigen 72-4 are jointly detected, so that the problem of low accuracy possibly caused by the fact that a marker detection method is mostly adopted in the existing early gastric cancer screening is solved.

Description

Technical Field

[0001] The present invention relates to the technical field of early gastric cancer screening, and specifically to a novel biomarker detection method for early gastric cancer screening. Background Art

[0002] Early gastric cancer refers to cancer tissue limited to the gastric mucosa layer and submucosa layer, regardless of whether there is lymph node metastasis. The symptoms of early gastric cancer are latent and easily overlooked. Some patients may only experience dull pain in the upper abdomen, fullness and discomfort, loss of appetite, indigestion, etc., similar to the symptoms of ordinary gastric diseases, and are prone to misdiagnosis or missed diagnosis. From the pathological type, early gastric cancer is mainly adenocarcinoma, which can be divided into papillary adenocarcinoma, tubular adenocarcinoma, mucinous adenocarcinoma, signet ring cell carcinoma, etc. Its incidence is related to multiple factors. Helicobacter pylori infection is an important cause. Long-term consumption of high-salt and pickled foods, smoking and excessive drinking, as well as having a family genetic history of gastric cancer, etc. will all increase the incidence risk. Early gastric cancer screening is of great significance and can improve the survival rate of patients. Common screening methods include gastroscopy, which can directly observe gastric mucosal lesions and, if necessary, take tissue biopsies to clarify the pathology; serological tests, such as the detection of pepsinogen, gastrin-17 biomarkers, which can assist in judging the state of the gastric mucosa. If detected and treated early, through minimally invasive means such as endoscopic mucosal resection or endoscopic submucosal dissection, most patients can achieve clinical cure, and the 5-year survival rate can reach more than 90%, and the quality of life can also be better guaranteed.

[0003] Most of the existing early gastric cancer screenings adopt a detection method using one biomarker. However, since the detection method using only one biomarker may produce errors or have certain limitations, it may cause the problem of low accuracy. Summary of the Invention

[0004] In view of the deficiencies of the prior art, the present invention provides a novel biomarker detection method for early gastric cancer screening, which solves the problem that most of the existing early gastric cancer screenings adopt a detection method using one biomarker. However, since the detection method using only one biomarker may produce errors or have certain limitations, it may cause the problem of low accuracy.

[0005] To achieve the above objectives, the present invention is realized through the following technical solutions: A novel biomarker detection method for early gastric cancer screening, comprising the following steps: S1. Preparation before detection: Detect pepsinogen in combination with gastrin-17, carcinoembryonic antigen, and carbohydrate antigen 72-4, and prepare the corresponding detection reagents and standards; S2. Sample collection: Use a disposable sterile blood collection needle and vacuum tube to draw 3-5 ml of venous blood through aseptic operation; S3. Sample processing and storage: Separate the serum by a centrifuge, and after the processing is completed, put the sample into a storage device for storage; S4, Detection operation: Preheat the detector and then calibrate the detector with the standard product, and detect the sample according to the detector operation manual; S5, Data analysis and result interpretation: Accurately record the detection data of the biomarker, and then compare and evaluate the detected value according to the standard range of the reference; S6, Disease prediction: Input the patient's detection data and clinical information into the big data platform, and establish a disease prediction model through big data analysis technology and machine learning algorithms to predict the risk of gastric cancer occurrence in the patient.

[0006] Preferably, in S1, the patient to be detected should not eat or drink more than 200 ml within 8 - 12 hours before the detection, and proton pump inhibitors, H2 receptor antagonists, and bismuth agents should be discontinued one week before the detection.

[0007] Preferably, in S2, the time period for sample collection is controlled from 8:00 am to 10:00 am.

[0008] Preferably, in S2, the positions for collecting venous blood samples include the median cubital vein, cephalic vein, and basilic vein.

[0009] Preferably, in S3, the rotation speed of the centrifuge is controlled at 3000 - 5000 r / min, the relative centrifugal force is controlled at 2000 - 4000 × g, the centrifugation time is controlled at 5 - 15 minutes, and the temperature is controlled at 4°C.

[0010] Preferably, in S3, the storage includes short - term storage, long - term storage, and ultra - long - term storage. For short - term storage, the temperature of the storage device is controlled at 2 - 8°C and the storage time is controlled at 12 - 24 hours. For long - term storage, the temperature of the storage device is controlled at - 20°C to - 40°C and the storage time is controlled at 6 - 12 months. For ultra - long - term storage, the temperature of the storage device is controlled at - 70°C to - 80°C and the storage time is controlled at 5 - 10 years.

[0011] Preferably, in S5, the results of pepsinogen, gastrin - 17, carcinoembryonic antigen, and carbohydrate antigen 72 - 4 are comprehensively compared.

[0012] Preferably, in S6, the clinical information includes the patient's age, gender, family medical history, eating habits, and Helicobacter pylori infection status.

[0013] The present invention provides a new biomarker detection method for early gastric cancer screening. It has the following beneficial effects: 1. In the present invention, by jointly detecting pepsinogen, gastrin-17, carcinoembryonic antigen, and carbohydrate antigen 72-4, it is possible to reflect the gastric health status from different perspectives using different markers. Multiple markers complement each other, reducing the errors caused by a single marker being interfered by various factors, improving the accuracy of early gastric cancer screening, and thus improving the existing detection methods for early gastric cancer that mostly use a single marker. However, since the detection method using only one marker may produce errors or have certain limitations, it may cause problems with low accuracy.

[0014] 2. In the present invention, by inputting the patient's test data and clinical information into a big data platform and using big data analysis technology and machine learning algorithms to establish a disease prediction model. This method comprehensively considers various factors and can more accurately predict the risk of gastric cancer in patients, providing a basis for early intervention and prevention. By including clinical information such as the patient's age, gender, family medical history, eating habits, and Helicobacter pylori infection status, the disease prediction model can more comprehensively consider individual differences and risk factors for disease onset, improving the accuracy and pertinence of risk prediction. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 is a flowchart of the method of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0016] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without making creative efforts shall fall within the protection scope of the present invention.

[0017] Please refer to the attached Figure 1 , an embodiment of the present invention provides a new biomarker detection method for early gastric cancer screening, which is characterized in that it includes the following steps: S1. Preparation before detection: Jointly detect pepsinogen, gastrin-17, carcinoembryonic antigen, and carbohydrate antigen 72-4, and prepare the corresponding detection reagents and standards. S2. Sample collection: Use a disposable sterile blood collection needle and vacuum tube to draw 3-5 ml of venous blood through aseptic operation. S3. Sample processing and storage: Separate the serum by a centrifuge, and store the sample in a storage device after the processing is completed. S4. Detection operation: Preheat the detector and then calibrate the detector with the standard, and detect the sample according to the operation manual of the detector. S5. Data analysis and result interpretation: Accurately record the detection data of the biomarker, and then compare and evaluate the detected value according to the standard range of the reference. S6, Disease prediction: Input the patient's test data and clinical information into the big data platform, and establish a disease prediction model through big data analysis technology and machine learning algorithms to predict the risk of gastric cancer occurrence in the patient.

[0018] In S1, the patient to be tested should not eat or drink more than 200 ml of water 8 - 12 hours before the test, and proton pump inhibitors, H2 receptor antagonists, and bismuth agents should be discontinued one week before the test.

[0019] Specifically, by not eating and drinking no more than 200 ml of water 8 - 12 hours before the test, it can ensure that the stomach is in a fasting state during the test. Eating will stimulate the secretion of gastrointestinal digestive juices, change the physiological state of the gastrointestinal tract, and may affect the secretion and release of pepsinogen and gastrin - 17 biomarkers, thereby interfering with the accuracy of the test results. By drinking no more than 200 ml of water, it can not only maintain the body's basic water requirements but also minimize the impact on the test indicators, ensuring that the test data can truly reflect the basic levels of relevant biomarkers in the patient's body. Proton pump inhibitors, H2 receptor antagonists, and bismuth agents will interfere with gastric acid secretion and the state of the gastric mucosa. Proton pump inhibitors and H2 receptor antagonists can inhibit gastric acid secretion, change the gastric pH value, and affect the conversion process of pepsinogen to pepsin in the stomach, resulting in abnormal fluctuations in the level of pepsinogen in the blood; bismuth agents may affect the metabolism and function of gastric mucosal cells and interfere with the secretion regulation of substances such as gastrin - 17. By discontinuing proton pump inhibitors, H2 receptor antagonists, and bismuth agents one week before the test, it can avoid the interference of drug factors on the test results, further ensure that the test results can accurately reflect the true pathophysiological state of the patient's own stomach, and improve the accuracy of early gastric cancer screening.

[0020] In S2, the time period for sample collection is controlled from 8:00 am to 10:00 am.

[0021] Specifically, various physiological functions of the human body show rhythmic changes throughout the day. From 8:00 am to 10:00 am, after a night's rest, the human body is in a relatively stable basal metabolic state. At this time, physiological activities such as hormone secretion and cell metabolism are relatively regular, and the levels of pepsinogen, gastrin - 17, carcinoembryonic antigen, and carbohydrate antigen 72 - 4 biomarkers are also relatively stable, with less interference from external factors such as diet, exercise, and emotion. By collecting samples from 8:00 am to 10:00 am, the content of the detected biomarkers can more truly reflect the basal level of the human body and reduce the differences in test results caused by physiological rhythm fluctuations. Fixing the sample collection time allows the tests of different patients to be carried out under similar physiological conditions, reduces the errors caused by differences in collection time, and enhances the comparability of test results.

[0022] In S2, the venous blood collection sites for sample collection include the median cubital vein, cephalic vein, and basilic vein.

[0023] Specifically, this facilitates operation. The median cubital vein, cephalic vein, and basilic vein are superficial in position, easy to observe and palpate for positioning. Medical staff can quickly and accurately find the blood vessels for puncture. Especially the median cubital vein, which is usually thick, straight, and fixed, with relatively less surrounding tissue. The feel during puncture is obvious, effectively reducing the puncture difficulty, reducing the pain caused to patients by repeated punctures, and at the same time improving the success rate and efficiency of blood collection. At the same time, this can ensure the sample quality. These veins have rich and relatively stable blood flow, and the collected blood samples can more accurately reflect the true situation in the patient's body. Compared with the veins in other parts, they are less affected by factors such as limb movement and external compression, avoiding a decline in sample quality due to poor blood flow or local blood component changes, and thus ensuring the accurate and reliable content of the pepsinogen, gastrin-17, carcinoembryonic antigen, and carbohydrate antigen 72-4 biomarkers detected, providing effective data support for early gastric cancer screening. At the same time, it can also reduce the risk of complications. Selecting the median cubital vein, cephalic vein, and basilic vein for blood collection can reduce the risk of complications during the blood collection process. The distribution of nerves and arteries around these veins is relatively clear, and medical staff are familiar with their anatomical structures, effectively avoiding nerves and arteries during puncture, reducing serious complications such as nerve injury and arterial puncture bleeding caused by accidental puncture, and ensuring the safety of patients.

[0024] In S3, control the rotation speed of the centrifuge at 3000 - 5000 r / min, control the relative centrifugal force at 2000 - 4000 × g, control the centrifugation time at 5 - 15 minutes, and control the temperature at 4°C.

[0025] Specifically, this can optimize the serum separation effect. By controlling the centrifuge speed at 3000 - 5000 r / min, the relative centrifugal force at 2000 - 4000 × g, and the centrifugation time at 5 - 15 minutes, efficient separation of blood cells and serum can be achieved. Within this parameter range, the centrifugal force is sufficient to quickly sediment the formed elements such as red blood cells and white blood cells in the blood, resulting in a clear stratification with the serum. If the speed, centrifugal force is too low or the time is too short, the blood cells may not sediment completely, leading to the presence of blood cell components in the serum and affecting the accuracy of subsequent test results. Conversely, if the parameters are too high or the time is too long, it may damage the blood cells, causing the release of intracellular components into the serum and interfering with the detection of target biomarkers. At the same time, this can maintain the stability of biomarkers. By controlling the centrifugation temperature at 4°C, the degradation rate of biomarkers in the sample can be effectively slowed down. Biomarkers such as pepsinogen, gastrin-17, carcinoembryonic antigen, and carbohydrate antigen 72-4 are mostly proteins or polypeptides, which are prone to denaturation, decomposition and other reactions at higher temperatures. The low-temperature environment of 4°C can reduce the activity of these biomolecules, inhibit the progress of related chemical reactions, and maximize the structural and functional integrity of biomarkers, ensuring accurate determination of their content in subsequent tests. This can also ensure the reliability of test results. By strictly controlling the centrifuge speed, centrifugal force, time and temperature, the consistency of sample processing conditions can be guaranteed each time, reducing detection errors caused by differences in the sample processing process. A standardized sample processing procedure helps to improve the repeatability and comparability of test results, making the test data at different times and batches more reliable, providing a solid guarantee for the accurate screening and diagnosis of early gastric cancer.

[0026] The storage in S3 includes short-term storage, long-term storage and ultra-long-term storage. For short-term storage, the temperature of the storage device is controlled at 2 - 8°C and the storage time is controlled at 12 - 24 hours. For long-term storage, the temperature of the storage device is controlled at -20°C to -40°C and the storage time is controlled at 6 - 12 months. For ultra-long-term storage, the temperature of the storage device is controlled at -70°C to -80°C and the storage time is controlled at 5 - 10 years.

[0027] Specifically, by storing the sample in an environment of 2-8°C for 12-24 hours in the short term, it is applicable to the situation where detection will be carried out in a short time. This temperature range can not only inhibit the growth and reproduction of microorganisms, reduce the risk of sample contamination, but also slow down the degradation rate of biomarkers to a certain extent. At the same time, compared with the storage conditions at lower temperatures, the environment of 2-8°C has relatively lower requirements for storage equipment, is more convenient to operate, can meet the needs of temporary storage of samples in the daily detection process of the laboratory, ensure that the changes in the properties and contents of biomarkers in the sample are minimized during the waiting for detection, and thus ensure that the detection results can accurately reflect the state of the sample at the time of collection. When storing for a long time, the temperature of the storage equipment is controlled at -20°C to -40°C, and the storage time is 6-12 months. In this temperature range, the activity of water molecules in the sample is significantly reduced, and the biochemical reaction rate is greatly slowed down, which can effectively inhibit the degradation, oxidation of biomarkers and the activities of microorganisms. For some samples that need to be detected multiple times within a period of time or are used for retrospective studies, this storage condition can not only maintain the relative stability of the sample, but also meet the subsequent detection needs within a certain period. Compared with ultra-long-term storage, the storage cost required for this temperature range is relatively low, which is applicable to the medium-term sample storage needs and balances the sample storage effect and the investment of laboratory resources. By storing the sample in an ultra-low temperature environment of -70°C to -80°C for 5-10 years, it is mainly used for situations where extremely high sample quality is required, long-term retention is needed for in-depth research or long-term monitoring. At such a low temperature, the biomolecules in the sample are almost in a "static" state, greatly reducing the structural changes and loss of activity of biomarkers, and can maintain the original state of the sample to the greatest extent. This is of great significance for studying the occurrence and development mechanism of gastric cancer, tracking the long-term change trend of diseases, and verifying new detection methods, etc., providing a reliable sample basis for long-term medical research and clinical practice.

[0028] In S5, the results of pepsinogen are comprehensively compared with those of gastrin-17, carcinoembryonic antigen, and carbohydrate antigen 72-4.

[0029] Specifically, this can achieve complementary advantages. Pepsinogen is divided into PGⅠ and PGⅡ, which can reflect the secretion function and atrophy of the gastric mucosa. Gastrin-17 is G-17, which can assist in judging the status of the gastric antrum mucosa and gastric acid secretion function. Carcinoembryonic antigen is CEA, and carbohydrate antigen 72-4 is CA72-4, which provides information from the perspective of tumor correlation. When abnormal pepsinogen indicates that there may be lesions in the gastric mucosa, combined with the changes in gastrin-17, it can further clarify whether the problem is in the gastric body or the gastric antrum; if the carcinoembryonic antigen or carbohydrate antigen 72-4 increases at the same time, the possibility of cancer is increased, the accuracy of early gastric cancer screening is improved, and the occurrence of missed diagnosis and misdiagnosis is reduced. If PGⅠ, the PGⅠ / PGⅡ ratio, G-17, CEA, and CA72-4 are all within the normal range, it indicates that there may be no obvious organic lesions in the stomach at present or the risk of canceration is relatively low, but the possibility of early micro-lesions cannot be completely excluded. When PGⅠ decreases, the PGⅠ / PGⅡ ratio decreases, and at the same time G-17 decreases, while CEA and CA72-4 are normal, it is mostly considered that there is atrophic gastritis in the gastric body or gastric antrum, and further gastroscopy examination is needed to clarify the diagnosis and regular monitoring. When PG is basically normal, G-17 increases, and CEA and CA72-4 increase slightly or are normal, it may indicate the presence of gastric antrum inflammation or Helicobacter pylori infection, and it is recommended to conduct relevant examinations such as Helicobacter pylori detection. If PG is abnormal and at the same time CEA and CA72-4 increase significantly, especially when CA72-4 > 10 U / mL and CEA > 10 ng / mL, regardless of whether G-17 is abnormal, gastric cancer is highly suspected, and gastroscopy examination and pathological biopsy should be carried out as soon as possible to clarify the diagnosis. For patients who have been diagnosed with gastric diseases, regular combined detection of these four indicators and observation of the dynamic changes of the indicators are helpful for evaluating the progression of the disease, treatment effect, and judging the prognosis. For example, during the treatment process, if CEA and CA72-4 gradually decline to normal and PG and G-17 improve, it indicates that the treatment is effective; on the contrary, if the indicators continue to increase, it may indicate disease recurrence or progression.

[0030] The clinical information in S6 includes the patient's age, gender, family history, eating habits, and Helicobacter pylori infection status.

[0031] Specifically, this enables accurate risk assessment. As people age, the risk of gastric cancer gradually increases, and there are differences in the cancer risk among people of different age groups. At the same time, the probability of men developing gastric cancer is relatively higher than that of women. Incorporating this information allows the disease prediction model to more accurately assess the individual's risk of gastric cancer based on the incidence characteristics of different ages and genders. This also enables consideration of genetic factors. If there is a gastric cancer patient in the family, the possibility of an individual carrying relevant susceptibility genes due to genetic factors increases, and the incidence risk is significantly higher than that of the general population. Understanding the family medical history can help the disease prediction model identify individuals with genetic susceptibility and assign corresponding weights when assessing risks, improving the ability to identify high-risk populations. It can also pay attention to the impact of lifestyle habits. Collecting information on eating habits helps the model comprehensively consider the impact of lifestyle on gastric health, thereby more accurately judging the likelihood of an individual developing gastric cancer.

[0032] Although the embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A novel biomarker detection method for early gastric cancer screening, characterized in that: It includes the following steps: S1. Preparation before detection: Combine the detection of pepsinogen with gastrin-17, carcinoembryonic antigen, and carbohydrate antigen 72-4, and prepare the corresponding detection reagents and standards. S2. Sample collection: Use a disposable sterile blood collection needle and vacuum tube to draw 3-5 ml of venous blood through aseptic operation. S3. Sample processing and storage: Separate the serum by a centrifuge, and after the processing is completed, place the sample in a storage device for storage. S4. Detection operation: Preheat the detector and then calibrate the detector with the standard, and detect the sample according to the operation manual of the detector. S5. Data analysis and result interpretation: Accurately record the detection data of the markers, and then compare and evaluate the detected values according to the standard range of the reference. S6. Disease prediction: Input the detection data and clinical information of the patient into the big data platform, and establish a disease prediction model through big data analysis technology and machine learning algorithms to predict the risk of gastric cancer occurrence in the patient.

2. The novel biomarker detection method for early gastric cancer screening according to claim 1, wherein: In S1, the patient to be detected should not eat or drink more than 200 ml within 8-12 hours before detection, and proton pump inhibitors, H2 receptor antagonists, and bismuth agents should be discontinued one week before detection.

3. The novel biomarker detection method for early gastric cancer screening according to claim 1, characterized in that: In S2, the time period for sample collection is controlled from 8:00 am to 10:00 am.

4. The novel biomarker detection method for early gastric cancer screening according to claim 1, characterized in that: In S2, the venous blood collection positions for sample collection include the median cubital vein, cephalic vein, and basilic vein.

5. The novel biomarker detection method for early gastric cancer screening according to claim 1, characterized in that: In S3, control the rotation speed of the centrifuge at 3000-5000 r / min, control the relative centrifugal force at 2000-4000×g, control the centrifugation time at 5-15 minutes, and control the temperature at 4°C.

6. The novel biomarker detection method for early gastric cancer screening according to claim 1, characterized in that: In S3, the storage includes short-term storage, long-term storage, and ultra-long-term storage. For short-term storage, control the temperature of the storage device at 2-8°C and the storage time at 12-24 hours. For long-term storage, control the temperature of the storage device at -20°C to -40°C and the storage time at 6-12 months. For ultra-long-term storage, control the temperature of the storage device at -70°C to -80°C and the storage time at 5-10 years.

7. The novel biomarker detection method for early gastric cancer screening according to claim 1, characterized in that: In S5, comprehensively compare the results of pepsinogen with gastrin-17, carcinoembryonic antigen, and carbohydrate antigen 72-4.

8. The novel biomarker detection method for early gastric cancer screening according to claim 1, characterized in that: In S6, the clinical information includes the patient's age, gender, family medical history, eating habits, and Helicobacter pylori infection status.

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