CN116559477A - Method for detecting eight indexes of stomach function at one time by self-help fingertip blood based on Internet - Google Patents
Method for detecting eight indexes of stomach function at one time by self-help fingertip blood based on Internet Download PDFInfo
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Abstract
The invention discloses a method for one-time detection of eight indexes of stomach function by self-help fingertip blood based on Internet. The method comprises the following steps: the method comprises the steps of collecting blood samples, inputting blood sample information by a mobile phone, carrying out blood sample delivery, preparing a fingertip pepsinogen sample to be detected and a helicobacter pylori sample to be detected, detecting the content of PGI and PG II and the value of PGI/PG II in a double-antibody sandwich mode, and judging whether the stomach function is normal or not; detecting IgG antibody of helicobacter pylori specific antigen CagA, vacA, ure and helicobacter pylori type I strain and helicobacter pylori type II strain in double antigen sandwich mode, and judging whether to infect helicobacter pylori; the user mobile phone inquires and reads the electronic report on line. The method can realize home self-help real-time detection, is not limited by factors such as environment, site, space distance, time, professional level of operators and the like, has the characteristics of high efficiency, convenience, accuracy, economy and the like, and can be used as a substitute solution of a health physical examination gastroscope detection project.
Description
Technical Field
The invention relates to a method for detecting stomach function indexes, in particular to a method for detecting eight stomach function indexes at one time by self-help fingertip blood based on the Internet.
Background
Serological detection is used as a non-invasive diagnosis method, and is simple to operate and high in acceptance. Serological detection ABC method is a method for detecting the occurrence risk of gastric cancer by combining pepsinogen with helicobacter pylori antibody.
The application value of pepsinogen in early diagnosis of gastric cancer is internationally recognized and is known as serological biopsy. When the gastric mucosa is changed, the PG content of blood is changed, so that the serum PG level can reflect the structure and function of the gastric mucosa, and is a detection index of stomach diseases such as superficial gastritis, erosive gastritis, gastric ulcer, duodenal ulcer, atrophic gastritis, gastric cancer and the like.
Helicobacter pylori has been shown to cause atrophic gastritis and intestinal metaplasia, both of which are precancerous lesions. A follow-up observation of up to 8 years for 118 patients post-radical helicobacter pylori has found that by successful radical treatment, the degree of antrum and gastric atrophy is significantly reduced, and the degree of gastric metaplasia is significantly reduced, whereas in patients not undergoing radical treatment, the degree of atrophy and metaplasia are not improved. Therefore, helicobacter pylori can be used as a dangerous index for gastric cancer occurrence, and the sensitivity of diagnosing gastric cancer is higher.
The combined detection of pepsinogen and helicobacter pylori has high clinical value, can comprehensively evaluate gastric mucosa atrophy, is a most practical screening method for further implementing gastroscopy on high-risk groups of gastric cancer, and improves the accuracy of gastric cancer diagnosis and screening.
At present, a venous blood sampling mode (blood sampling amount is 2-3 ml) is generally used for detecting pepsinogen indexes in clinic, and a carbon 13 and carbon 14 expiration test method is used for detecting helicobacter pylori antibodies. The methods for detecting pepsinogen index and helicobacter pylori antibody by carbon 13 and carbon 14 through venous blood drawing are carried out by special medical institutions, so that time and labor are consumed, the detection cost is high, and the methods are limited by factors such as time, place, professionals and the like. In addition, the carbon 13 and carbon 14 exhalations test method can only detect positive or negative conditions of helicobacter pylori urease antibodies, and cannot be used for typing helicobacter pylori (different clinical treatment medicines of different subtypes are different).
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a method for detecting eight indexes of the stomach function at one time by self-help fingertip blood based on the Internet.
In order to achieve the above purpose, the technical scheme adopted by the invention comprises the steps of blood sample collection, blood sample information input by a mobile phone, blood sample delivery, blood sample registration, preparation of a detection sample, sample detection and detection report inquiry, and is characterized by comprising the following steps:
1) Collecting a blood sample by using a blood collecting card to collect fingertip blood;
2) Cell phone for inputting blood sample information
2.1 Scanning a two-dimensional code printed on the blood sampling card by using a mobile phone APP, and paying attention to public numbers;
2.2 Clicking a bar code button on an APP sample registration interface, and scanning a bar code printed on a blood sampling card by a mobile phone to finish personal information registration of a tested person;
2.3 Clicking a submit button on the sample registration interface, and selecting a call express button;
3) Collecting a blood sampling card for collecting fingertip blood at the upper gate of a blood sample delivery and inspection express delivery person, and sending the fingertip blood to a designated detection unit for free;
4) Registering the blood sample by scanning a bar code on the blood collection card;
5) Preparation of test samples
5.1 Using a puncher to punch 18 round holes at the dried blood spots on the blood sampling card;
5.2 Placing 4 round paper sheets with dry blood spots into a centrifuge tube, adding 150 microliters of pepsinogen sample diluent, and re-dissolving in a refrigerator at 2-8 ℃ for 16 hours to obtain a fingertip pepsinogen sample to be detected;
placing 14 circular paper sheets with dried blood spots into another separation tube, adding 300 microliter helicobacter pylori sample diluent, and re-dissolving in a refrigerator at 2-8 ℃ for 16 hours to obtain helicobacter pylori samples to be detected;
6) Sample detection
6.1 Quantitative detection of PGI and PG II content and PGI/PG II value in the sample to be detected of fingertip pepsinogen in a double-antibody sandwich mode by utilizing a fingertip pepsinogen detection reagent and a full-automatic quantum dot fluorescence immunoassay instrument, comparing the obtained detection index with a serum normal index, and judging whether the stomach function is normal or not;
6.2 Using helicobacter pylori IgG antibody detection kit and biochip reader, qualitatively detecting IgG antibody of helicobacter pylori specific antigen CagA, vacA, ure, helicobacter pylori type I strain and helicobacter pylori type II strain according to double antigen sandwich mode, comparing the obtained detection index with normal index, and judging whether helicobacter pylori is infected or not and strain typing (type I or type II):
if the detection values of CagA, vacA, ure are smaller than or equal to the corresponding positive judgment values, the three detection items are negative, which means that no helicobacter pylori antibody exists in the detected sample;
if one or more than one detection value of CagA, vacA, ure is greater than the corresponding positive judgment value, the helicobacter pylori IgG antibody is positive in detection, and the helicobacter pylori is present or once infected, and the helicobacter pylori can be regarded as present infection for untreated people;
7) The detection report can be inquired by inquiring the bar code on the blood sampling card 'stub' of the examinee through the mobile phone APP scanning.
In the technical scheme, the fingertip pepsinogen reagent comprises a detection card and a drying agent. The detection card consists of a test strip fixed in a plastic shell, wherein the test strip consists of a sample pad, a quantum dot marker pad, a nitrocellulose membrane, a buffer pad and a water absorption pad which are sequentially fixed on a polyvinyl chloride plate; the sample pad, the quantum dot marker pad and the nitrocellulose membrane are sequentially overlapped along one end of the polyvinyl chloride plate, and the water absorption pad, the buffer pad and the nitrocellulose membrane are sequentially overlapped along the other end of the polyvinyl chloride plate.
The quantum dot label pad contains an antibody 1 and an antibody 2, wherein the antibody 1 is a 0.4mg/ml mouse anti-human pepsinogen I antibody quantum dot conjugate, and the antibody 2 is a 0.4mg/ml mouse anti-human pepsinogen II antibody quantum dot conjugate; the nitrocellulose membrane is sprayed with a detection T line and a quality control C line, the detection T line is coated with an antibody 3 and an antibody 4, the antibody 3 is a 1mg/ml mouse anti-human pepsinogen I monoclonal antibody, and the antibody 4 is a 1mg/ml mouse anti-human pepsinogen II monoclonal antibody; detecting C line coated with pepsinogen I reference protein 0.1mg/ml and pepsinogen II reference protein 0.1 mg/ml.
In the technical scheme, the helicobacter pylori IgG antibody detection kit comprises a helicobacter pylori IgG antibody detection chip, a washing liquid, a color developing agent II and a sample diluent, wherein the helicobacter pylori IgG antibody detection chip is a protein chip obtained by fixing human immunoglobulin expressed by a genetic engineering technology, helicobacter pylori specific CagA recombinant antigen, helicobacter pylori specific VacA recombinant antigen, helicobacter pylori specific Ure recombinant antigen and pRSET-A plasmid transfected BL21 thallus lysate on a nitrocellulose membrane in a microarray mode.
The pRSET-A plasmid transfected BL21 thallus lysate is escherichia coli BL21 type, the color developing agent II is a staphylococcus A protein marker, the main components of the washing solution are TBE and saturated ammonium sulfate solution, the concentration of the prepared Tris buffer solution is 0.05mol/L, the main components of the sample diluent are 0.01M phosphate buffer solution, and the concentration of the prepared KH2PO4, na2HPO4.12H2O, naCL and KC1 is 0.01mol/L.
Finger tip pepsinogen test principle:
the fingertip blood detection pepsinogen sample is upwards diffused by capillary force on the sample pad, and PGI (pepsinogen I) and PG II (pepsinogen II) in the sample are respectively combined with an antibody 1 (a mouse anti-human pepsinogen I antibody quantum dot conjugate) and an antibody 2 (a mouse anti-human pepsinogen II antibody quantum dot conjugate) on the label pad to form a quantum dot labeled antibody-antigen complex 1 and a quantum dot labeled antibody-antigen complex 2 when passing through the label pad; the quantum dot labeled antibody-antigen complex 1 and the quantum dot labeled antibody-antigen complex 2 are continuously diffused onto a nitrocellulose membrane along with a sample, are intercepted and captured by detection T lines coated with an antibody 3 (a mouse anti-human pepsinogen I monoclonal antibody) and an antibody 4 (a mouse anti-human pepsinogen II monoclonal antibody), and respectively form an immune complex 1 of the quantum dot labeled antibody-antigen-coated antibody and an immune complex 2 of the quantum dot labeled antibody-antigen-coated antibody. The non-intercepted quantum dot labeled antibody-antigen complex 1 and the quantum dot labeled antibody-antigen complex 2 continue to ascend and are respectively combined with pepsinogen I internal reference protein and pepsinogen II internal reference protein coated by a quality control C line to indicate that the reaction is completed. And generating a fluorescence detection signal by exciting the quantum dots, and obtaining a quantitative detection result by using a quantum dot fluorescence immunoassay analyzer. The content of pepsinogen I and pepsinogen II in the sample to be detected by fingertip blood detection pepsinogen is in linear relation with the intensity of fluorescence detection signals in a certain range.
Helicobacter pylori test principle:
the specific IgG antibody in the sample to be detected and the antigen on the helicobacter pylori IgG antibody detection chip generate specific immune binding reaction, an antigen-antibody immune complex is formed on an NC film (nitrocellulose film), and then the labeled staphylococcus A protein is specifically bound with the antigen-antibody immune complex, finally macroscopic red spots appear, and the detection result is qualitatively analyzed by a biochip reader.
Compared with the prior art, the invention organically combines the Internet with fingertip blood serological index detection and helicobacter pylori antibody detection by adopting the technical scheme, so the invention has the following advantages:
1) Collecting fingertip blood to finish detection of eight stomach function indexes of pepsinogen I (PG I), pepsinogen II (PG II), pepsinogen I (PG I)/pepsinogen II (PGR), cytotoxin related protein A antibody (CagA), vacuolar toxin related protein A antibody (VacA), urease antibody (Ure), helicobacter pylori type I strain and helicobacter pylori type II strain at one time;
2) The helicobacter pylori can be accurately typed, so as to guide accurate therapeutic medication;
3) The self-help detection at home can be realized, the application scene of gastric cancer screening is widened, the detection of medical treatment in hospitals is not needed, and a great amount of time, energy and cost are saved for patients. The method has the characteristics of high efficiency, convenience, accuracy, economy, no limitation by factors such as environment, site, space distance, time, professional level of operators and the like, and can be used as a new means for screening gastric cancer in health examination.
Drawings
FIG. 1 is a schematic view of a blood collection card according to the present invention;
FIG. 2 is a schematic diagram of the structure of a fingertip pepsinogen reagent detection card according to the present invention;
FIG. 3 is a standard graph of fingertip pepsinogen PGI;
FIG. 4 is a standard graph of fingertip pepsinogen PGII;
FIG. 5 is a graph comparing clinical data of fingertip pepsinogen I (PGI) and Atlantic chemiluminescent (serum) pepsinogen I (PGI);
FIG. 6 is a comparison of clinical data of fingertip pepsinogen II (PGII) and Atlantic chemiluminescent (serum) pepsinogen II (PGII).
In the figure: the blood sampling card comprises a blood sampling card body 1, a recording unit 2, a bar code 3, a sample unit 4, a sampling window 5, a protective sleeve 6, a blood sampling filter paper sheet 7, viscose 8, a two-dimensional code 9, a stub unit 10, a sample pad 11, a quantum dot marker pad 12, a nitrocellulose membrane 13, a buffer pad 14, a water absorption pad 15, a polyvinyl chloride plate 16, a quality control C line 17 and a detection T line 18.
By detecting PGI and PGII of fingertip blood of 486 testers, a standard curve chart of PGI of fingertip pepsinogen (see figure 3 for details) and a standard curve chart of PGII of fingertip pepsinogen (see figure 4 for details) are obtained. The linear analysis results show that each concentration is linearly related to the detection value within the range of 0-400 ng/mL, and is Y=0.0379 x-0.385 and R 2
＝0.9971；Y＝0.0307x+0.1117，R 2 ＝0.9867。
And (3) drawing a standard curve chart: and measuring quality control substances with different concentrations by using a fingertip pepsinogen detection reagent, and drawing a standard curve by taking the concentration of the quality control substances as an abscissa and the ratio of fluorescent signals as an ordinate. Drawing: adding PGI quality control substances (370 ug/mL, 270ug/mL, 120ug/mL, 100ug/mL, 70ug/mL, 20ug/mL and 2 ug/mL) with different concentrations and PGII quality control substances (50 ug/mL, 30ug/mL, 20ug/mL, 15ug/mL, 10ug/mL, 5ug/mL and 2 ug/mL) with different concentrations into a fingertip pepsinogen detection kit, respectively diluting the PGI and PGII with fingertip blood to obtain 1000ug/mL quality control substance mother liquor, diluting the sample mother liquor with fingertip blood to different quality control substance concentrations, adding sample diluent, reading C, T line fluorescence signals and C/T values through a full-automatic quantum dot fluorescence immunoassay analyzer, and drawing standard curves according to PGI and PGII quality control substance concentrations and sample signals T1/C and T2/C average value.
And (3) precision inspection: three batches of PG I/PG II fingertip pepsinogen detection reagents are taken, 100ug/mL and 70ug/mL of repeated property control products are respectively detected by the PG I quality control products, 10 times of repeated property control products are parallelly detected by each batch of fingertip pepsinogen detection reagent, the CV (constant velocity) of the 100ug/mL concentration of three batches is respectively 4.23%, 6.07% and 4.96%, the CV (constant velocity) of the three batches is 5.08%, the CV (constant velocity) of the 70ug/mL concentration of three batches is respectively 6.31%, 7.03% and 5.51%, and the CV (constant velocity) of the three batches is 6.28% and is within 10%. The PG II quality control product is used for respectively detecting 15ug/mL and 10ug/mL of repetitive quality control product, each batch of fingertip pepsinogen detection reagent is parallelly detected for 10 times by the repetitive quality control product, CV (constant velocity) in three batches with the concentration of 15ug/mL is respectively 5.15%, 6.15% and 6.97%, CV (constant velocity) between three batches is 6.09%, CV (constant velocity) in three batches with the concentration of 10ug/mL is respectively 3.29%, 8.08% and 8.76%, CV (constant velocity) between three batches is 6.71%, and CV between three batches is within 10%.
FIG. 5 and FIG. 6 are graphs showing that the correlation between the clinical results of the finger tip pepsinogen reagent PGI and PGII and the Yaban chemiluminescent serum reagent PGI and PGII, and that the correlation between the two reagents is good when 486 clinical serum samples are detected in parallel 2 ＝0.9973，PGII R 2 0.9972, the average relative deviation is less than 10%, and the result meets the clinical analysis requirement, thereby achieving the clinical use effect.
Detailed Description
For the purpose of clearly describing the method of the present invention, it is necessary to first describe the structure of the blood collection card (improved based on CN201782762U, CN 204575377U) according to the present invention
The structure is described in a simple way:
as shown in fig. 1: the blood sampling card is composed of a blood sampling card body 1 with a protective sleeve 6 at the upper half part and a blood sampling filter paper sheet 7 which is enclosed in the protective sleeve by an adhesive 8. Five circular sampling windows 5 are respectively arranged at the corresponding positions of the front and the back of the protective sleeve 6, and two shearing lines (not shown in the figure) for dividing the blood sampling card body into a sample connector 4, a record connector 2 and a stub connector 10 from top to bottom are printed on the surface of the blood sampling card body 1. The bar codes 3 are printed on the surfaces of the sample couplet 4, the record couplet 2 and the stub couplet 10, and the two-dimensional codes 9 are printed on the surfaces of the stub couplet 10.
The process according to the invention is further described below with reference to the accompanying drawings and to specific examples:
examples
1) The blood sample collection comprises the steps of massaging the finger to make the finger congestion, wiping the tail end of the finger with alcohol cotton for disinfection, squeezing and pressing the blood sampling finger, tightly attaching the head of the blood sampling device to the abdomen of the finger and pressing the blood sampling device with force; after bleeding, wiping out first drop of blood by using alcohol cotton, extruding the drop of blood into soybean size by a continuous extruding-loosening-continuous extruding mode, and then respectively dripping the blood onto the blood sampling filter paper sheet 7 of the blood sampling card sampling window 5; 5 drops of blood (about 0.3 ml) are collected once, and the woundplast is used for stopping bleeding after the blood collection is finished;
2) Cell phone for inputting blood sample information
2.1 A mobile phone APP is used for scanning a two-dimensional code 9 printed on a blood sampling card special for fingertip blood to pay attention to public numbers;
2.2 Clicking a 'bar code' button on a sample registration interface, inputting a bar code 3 printed on a finger-tip blood special blood sampling card in a scanning mode, and filling in personal information such as the name, age, sex, contact telephone and the like of the testee;
2.3 Clicking a submit button on the registration interface, selecting a call express button;
3) Collecting a blood sampling card for collecting fingertip blood at the upper gate of a blood sample delivery and inspection express delivery person, and sending the fingertip blood to a designated detection unit for free;
4) The blood sample registration detection unit can register the received blood sampling card by scanning the bar code 3 on the blood sampling card sample unit 4;
5) Preparation of test samples
5.1 Using a puncher to punch 18 round holes with the diameter of 3 mm at the sampling window 5 of the blood sampling card;
5.2 Placing 4 round paper sheets with dry blood spots into a centrifuge tube, adding 150 microliters of pepsinogen sample diluent, and re-dissolving in a refrigerator at 2-8 ℃ for 16 hours to obtain a fingertip pepsinogen sample to be detected;
placing 14 circular paper sheets with dried blood spots into another separation tube, adding 300 microliter helicobacter pylori sample diluent, and re-dissolving in a refrigerator at 2-8 ℃ for 16 hours to obtain helicobacter pylori samples to be detected;
6) Sample detection
6.1 Quantitative detection of PGI and PG II content and PGI/PG II value in the sample to be detected by using the fingertip pepsinogen detection reagent and the full-automatic quantum dot fluorescence immunoassay instrument in a double-antibody sandwich mode; the specific operation steps are as follows:
6.1.1 Taking out the finger tip pepsinogen sample prepared in the step 5), and standing for 30 minutes at room temperature;
6.1.2 Inserting the detection card into the full-automatic quantum dot fluorescence immunoassay analyzer, extending the sample to be detected of the fingertip pepsinogen to the sample adding needle of the full-automatic quantum dot fluorescence immunoassay analyzer, immersing the tail end of the needle point into the liquid surface of the sample, pressing a detection key, waiting for the sample adding needle to leave the detection card upwards and then removing, and obtaining a detection index;
6.1.3 Comparing the obtained detection index with serum normal index (PGI is more than or equal to 70ng/ml, PG II is less than 150ng/ml, and PGI/PG II is more than or equal to 3) to judge whether the stomach function is normal;
6.2 IgG antibody of helicobacter pylori heterologous antigen CagA, vacA, ure, helicobacter pylori type I strain and helicobacter pylori type II strain are qualitatively detected by using helicobacter pylori IgG antibody detection kit and biochip reader according to double antigen sandwich method; the specific operation steps are as follows:
6.2.1 Taking out the helicobacter pylori to-be-detected sample prepared in the step 5), and standing for 30 minutes at room temperature;
6.2.2 A, wetting a film, namely adding 4 drops of washing liquid on the film surface of a detection window of the helicobacter pylori IgG antibody detection chip, and wetting the film surface;
6.2.3 Sampling 200 microliters of helicobacter pylori to-be-detected sample is added on the membrane surface;
6.2.4 Washing the helicobacter pylori to be detected sample to fully permeate the membrane surface, and then adding 6 drops of washing liquid to the membrane surface;
6.2.5 After the washing liquid fully permeates the membrane surface, 6 drops of the color developing agent II are taken and added to the membrane surface;
6.2.6 After the color developing agent II fully permeates the membrane surface, 6 drops of washing liquid are taken and added to the membrane surface;
6.2.7 After the washing liquid fully permeates the membrane surface, observing the color development conditions of the reaction control probe and the negative reaction probe; if the helicobacter pylori detection chip is effective, placing the helicobacter pylori detection chip into a sample tray of a chip reader for detecting CagA, vacA, ure, helicobacter pylori type I strain and helicobacter pylori type II strain;
6.2.8 Positive determination
The detection value of CagA, vacA, ure was compared with positive judgment values (CagA > 8, vacA > 8, ure > 8):
if the detection value of CagA, vacA, ure is smaller than or equal to the positive judgment value, all three detection items are negative, which means that no helicobacter pylori antibody exists in the detected sample;
if one or more than one detection value of CagA, vacA, ure is greater than the positive judgment value, the helicobacter pylori IgG antibody is positive in detection, and the helicobacter pylori is present or once infected, and the helicobacter pylori can be regarded as present infection for untreated people;
7) The detection report can be inquired by inquiring the bar code on the blood sampling card 'stub' of the examinee through the mobile phone APP scanning.
The structure of the test card for the fingertip pepsinogen reagent in the above embodiment is shown in fig. 2: it is constituted by a test strip fixed in a plastic housing (not shown in the figure) constituted by a sample pad 11, a quantum dot marker pad 12, a nitrocellulose membrane 13, a buffer pad 14 and a water absorbing pad 15, which are sequentially fixed on a polyvinyl chloride plate 16. The sample pad 11, the quantum dot marker pad 12, and the nitrocellulose membrane 13 are sequentially overlapped along one end of the polyvinyl chloride plate 16 (i.e., sequentially overlapped from left to right), and the water absorbing pad 15, the buffer pad 14, and the nitrocellulose membrane 13 are sequentially overlapped along the other end of the polyvinyl chloride plate 16 (i.e., sequentially overlapped from right to left).
The quantum dot label pad 12 contains an antibody 1 and an antibody 2, wherein the antibody 1 is a 0.4mg/ml mouse anti-human pepsinogen I antibody quantum dot conjugate, and the antibody 2 is a 0.4mg/ml mouse anti-human pepsinogen II antibody quantum dot conjugate. The nitrocellulose membrane 13 is sprayed with a detection T line 18 and a quality control C line 17, the detection T line 18 is coated with an antibody 3 and an antibody 4, the antibody 3 is a 1mg/ml mouse anti-human pepsinogen I monoclonal antibody, and the antibody 4 is a 1mg/ml mouse anti-human pepsinogen II monoclonal antibody; detection line 17 was coated with 0.1mg/ml pepsinogen I and 0.1mg/ml pepsinogen II.
The helicobacter pylori IgG antibody detection kit related in the embodiment comprises a helicobacter pylori IgG antibody detection chip, a washing liquid, a color developing agent II and a sample diluent, wherein the helicobacter pylori IgG antibody detection chip is a protein chip obtained by fixing human immunoglobulin expressed by a genetic engineering technology, helicobacter pylori specific CagA recombinant antigen, helicobacter pylori specific VacA recombinant antigen, helicobacter pylori specific Ure recombinant antigen and pRSET-A plasmid transfected BL21 thallus lysate on a nitrocellulose membrane in a microarray mode.
The pRSET-A plasmid transfected BL21 thallus lysate is escherichia coli BL21 type, the color developing agent II is a staphylococcus A protein marker, the main components of the washing solution are TBE and saturated ammonium sulfate solution, wherein the concentration of the prepared Tris buffer solution is 0.05mol/L, the main components of the sample diluent are 0.01M phosphate buffer solution, and the concentration of the prepared KH2PO4, na2HPO4.12H2O, naCL and KC1 is 0.01mol/L.
The following are data collected by physical examination of 1633 community persons (aged 28-79 years) using the method of the present invention for 5-11 months 2022: pepsinogen and helicobacter pylori are negative 978 people, the helicobacter pylori infection is positive 655 people, 40% is occupied, wherein the I type 430 people and the II type 225 people indicate the people with stomach disease risks, 36 precancerous lesions (the precancerous lesions are sequentially divided into chronic atrophic gastritis, intestinal metaplasia, atypical hyperplasia and severe atypical hyperplasia from light to heavy) are found through gastroscopy, the samples are 1627, the samples are 6, the accurate total coincidence rate is 99%, and the negative predictive value is 99.4%.
Table 1: comparison of infection Rate and infection typing of helicobacter pylori in each group
Table 2: comparison of serum PGI, PG II and PGR of each group
| Group of | Number of examples | PGI(ng/ml) | PGⅡ(ng/ml) | PGR |
| Atrophic gastritis | 13 | 55.5±7.2 | 24.6±2.1 | 1.9±1.1 |
| Intestinal metaplasia | 10 | 53.6±5.6 | 28.6±2.4 | 1.7±1.1 |
| Atypical hyperplasia | 7 | 45.3±4.2 | 32.5±2.3 | 1.5±0.8 |
| Severe atypical hyperplasia | 6 | 42.1±3.8 | 37.8±2.1 | 1.2±0.5 |
The method of the invention is adopted below, and a plurality of groups of detection typical cases are carried out by taking Pepsinogen (PG) and Helicobacter Pylori (HP) antibody combination method (namely ABC method) as evaluation criteria in Chinese medical society's stomach cancer clinical diagnosis and treatment guide (2021 edition).
Case 1, zhang Mouzhong, men, 59 years, 2022, 6 and 23 days, tested eight indices of gastric function using the method of the invention:
the stomach function health risk grade is evaluated as grade C, and has the risks of infection of helicobacter pylori, chronic gastritis, gastric ulcer, erosive gastritis and the like. The gastroscopy of the tester at 2022, 7 and 1 days gave the following results: stomach Dou Xirou, chronic non-atrophic gastritis, helicobacter pylori infection.
Case 2, zhang Mou, women, 39 years, 2022, 5 months and 19 days the method of the invention was used to detect eight indices of gastric function:
the stomach function health risk grade is evaluated as grade D, the PGI value and the PGR value are lower than the normal values, and the risks of early gastric ulcer, erosive gastritis and the like are presented, so that the gastroscopy in the hospital is recommended. The gastroscopy results of the inspector 2022, 5 and 25 days were: acute hemorrhagic gastritis.
Case 3, yang Mouping, female, 59 years, 2022, 8 months, 26 days, eight indices of gastric function were tested using the method of the invention:
the stomach function health risk grade is evaluated as grade B, helicobacter pylori antibody is positive, the type I is typed, pepsinogen II value is abnormal, and gastroscopy is recommended. The gastroscopy results of the inspector 2022, 9 and 2 days were: non-atrophic gastritis, helicobacter pylori infection.
Case 4, li Mou, female, 59 years, 2022, 11, 23 days eight indices of gastric function were tested using the method of the invention:
gastric function health risk grade was assessed as grade D, helicobacter pylori antibody negative, pepsinogen PGI number abnormal, PGR number abnormal, suggesting gastroscopy. The gastroscopy results of the test person at 2022, 12, 28 were: chronic atrophic gastritis.
Claims (4)
1. The method for one-time detection of eight indexes of the stomach function by self-service fingertip blood based on the Internet comprises the steps of blood sample collection, blood sample information input by a mobile phone, blood sample delivery and detection, blood sample registration, preparation of a detection sample, sample detection and detection report query, and is characterized by comprising the following steps:
1) Collecting a blood sample by using a blood collecting card to collect fingertip blood;
2) Cell phone for inputting blood sample information
2.1 Scanning a two-dimensional code printed on the blood sampling card by using a mobile phone APP, and paying attention to public numbers;
2.2 Clicking a bar code button on an APP sample registration interface, and scanning a bar code printed on a blood sampling card by a mobile phone to finish personal information registration of a tested person;
2.3 Clicking a submit button on the sample registration interface, and selecting a call express button;
3) Collecting a blood sampling card for collecting fingertip blood at the upper gate of a blood sample delivery and detection express delivery person, and sending the fingertip blood to a designated detection unit for free;
4) Registering the blood sample by scanning a bar code on the blood collection card;
5) Preparation of test samples
5.1 Using a puncher to punch 18 round holes at the dried blood spots on the blood sampling card;
5.2 Placing 4 round paper sheets with dry blood spots into a centrifuge tube, adding 150 microliters of pepsinogen sample diluent, and re-dissolving in a refrigerator at 2-8 ℃ for 16 hours to obtain a fingertip pepsinogen sample to be detected;
placing 14 circular paper sheets with dried blood spots into another separation tube, adding 300 microliter helicobacter pylori sample diluent, and re-dissolving in a refrigerator at 2-8 ℃ for 16 hours to obtain helicobacter pylori samples to be detected;
6) Sample detection
6.1 Using fingertip pepsinogen detection reagent and full-automatic quantum dot fluorescence immunoassay analyzer to quantitatively detect the content of PGI and PG II and the value of PGI/PG II in pepsinogen to-be-detected sample by fingertip blood according to a double-antibody sandwich mode, comparing the obtained detection index with serum blood normal index and judging whether the stomach function is normal;
6.2 Using helicobacter pylori IgG antibody detection kit and biochip reader, qualitatively detecting IgG antibody of helicobacter pylori specific antigen CagA, vacA, ure, helicobacter pylori type I strain and helicobacter pylori type II strain according to double antigen sandwich mode, comparing the obtained detection index with normal index, and judging whether helicobacter pylori is infected or not and strain typing:
if the detection values of CagA, vacA, ure are smaller than or equal to the corresponding positive judgment values, the three detection items are negative, which means that no helicobacter pylori antibody exists in the detected sample;
if one or more than one detection value of CagA, vacA, ure is greater than the corresponding positive judgment value, the helicobacter pylori IgG antibody is positive in detection, and the helicobacter pylori is present or once infected, and the helicobacter pylori can be regarded as present infection for untreated people;
the detection report can be inquired by inquiring the bar code on the blood sampling card 'stub' of the examinee through the mobile phone APP scanning.
2. The internet-based self-help fingertip blood one-time stomach function eight index detection method of claim 1, wherein: the fingertip pepsinogen detection reagent comprises a detection card and a drying agent, wherein the detection card is composed of a test strip fixed in a plastic shell, and the test strip is composed of a sample pad (11), a quantum dot marker pad (12), a nitrocellulose membrane (13), a buffer pad (14) and a water absorption pad (15) which are sequentially fixed on a polyvinyl chloride plate (16); the sample pad (11), the quantum dot marker pad (12) and the nitrocellulose membrane (13) are sequentially overlapped along one end of the polyvinyl chloride plate (16), and the water absorption pad (15), the buffer pad (14) and the nitrocellulose membrane (13) are sequentially overlapped along the other end of the polyvinyl chloride plate (16);
the quantum dot marker pad (12) contains an antibody 1 and an antibody 2, wherein the antibody 1 is a 0.4mg/ml mouse anti-human pepsinogen I antibody quantum dot conjugate, and the antibody 2 is a 0.4mg/ml mouse anti-human pepsinogen II antibody quantum dot conjugate;
the nitrocellulose membrane (13) is sprayed with a detection T line (18) and a quality control C line (17), the detection T line (18) is coated with an antibody 3 and an antibody 4, the antibody 3 is a 1mg/ml mouse anti-human pepsinogen I monoclonal antibody, and the antibody 4 is a 1mg/ml mouse anti-human pepsinogen II monoclonal antibody; the test line C (17) was coated with 0.1mg/ml pepsinogen I and 0.1mg/ml pepsinogen II.
3. The internet-based self-help fingertip blood one-time stomach function eight index detection method of claim 1, wherein: the helicobacter pylori IgG antibody detection kit comprises a helicobacter pylori IgG antibody detection chip, a washing liquid, a color developing agent II and a sample diluent, wherein the helicobacter pylori IgG antibody detection chip is a protein chip obtained by fixing human immunoglobulin expressed by a genetic engineering technology, helicobacter pylori specific CagA recombinant antigen, helicobacter pylori specific VacA recombinant antigen, helicobacter pylori specific Ure recombinant antigen and pRSET-A plasmid transfected BL21 thallus lysate on a nitrocellulose membrane in a microarray mode.
4. The internet-based self-help fingertip blood one-time stomach function eight index detection method of claim 3, wherein: the pRSET-A plasmid transfected BL21 thallus lysate is escherichia coli BL21 type, the color developing agent II is a staphylococcus A protein marker, the main components of the washing solution are TBE and saturated ammonium sulfate solution, wherein the concentration of the prepared Tris buffer solution is 0.05mol/L, the main components of the sample diluent are 0.01M phosphate buffer solution, and the concentration of the prepared KH2PO4, na2HPO4.12H2O, naCL and KC1 is 0.01mol/L.
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