CN111366724B

CN111366724B - Rare earth detection test strip, detection card, kit and preparation method for atrophic gastritis

Info

Publication number: CN111366724B
Application number: CN202010192811.1A
Authority: CN
Inventors: 张芳榕; 吴文源
Original assignee: Xiamen Amonmed Biotechnology Co ltd
Current assignee: Xiamen Amonmed Biotechnology Co ltd
Priority date: 2020-03-18
Filing date: 2020-03-18
Publication date: 2022-09-30
Anticipated expiration: 2040-03-18
Also published as: CN111366724A

Abstract

The invention discloses a atrophic gastritis rare earth detection test strip, a detection card, a kit and a preparation method, wherein the atrophic gastritis rare earth detection test strip comprises a bottom lining, and a sample pad, a combination pad, a nitrocellulose membrane and absorbent paper which are sequentially lapped on the bottom lining along the length direction; rare earth nanometer fluorescent microspheres marked with PG I, PG II and rabbit IgG antibodies are sprayed on the bonding pad; the nitrocellulose membrane is sequentially scribed through monoclonal antibodies corresponding to PG I, PG II and rabbit IgG antibodies, and the monoclonal antibodies are respectively used as a first detection line, a second detection line and a quality control line; the first detection line is close to the combination pad, and the quality control line is close to the absorbent paper. The invention has the characteristics of wide detection range, high sensitivity, high accuracy, quick, simple and convenient detection and the like, and the detection result of the invention can help doctors to accurately diagnose atrophic gastritis at an early stage.

Description

Atrophic gastritis rare earth detection test strip, detection card, kit and preparation method

Technical Field

The invention relates to a rare earth test strip for atrophic gastritis, a test card, a kit and a preparation method.

Background

Pepsinogen (PG) consists of 375 amino acids, has a molecular weight of about 42k Da, is synthesized and secreted mainly by gastric mucosal tissues, and can be activated into pepsin with a digestive function under the stimulation of gastric acid. The sources of PG I and PG II are different, PG I is mainly secreted by the main cells of the fundus ventriculi and the mucous cervical cells; meanwhile, PG II is derived from almost all gastric gland cells, such as gastric cardia gland, fundus gland, antrum pylorus gland cells, and even further comprises distal duodenal Brunner gland cells. Normally, most of the synthesized PG enters the gastric cavity directly, and only a few (about 1%) permeate the capillaries of the gastric mucosa into the blood circulation and remain stable. The PG level of serum reflects the forms and functions of gastric mucosa at different parts, PG I is a pointer for detecting the functions of gastric acid gland cells, and PG I is increased due to the increase of gastric acid secretion, decreased in secretion or decreased in atrophy of gastric mucosa glands; PG II has a large correlation with the pathological changes of the gastric fundus mucosa (relative to the gastric antral mucosa), and the increase of PG II is related to atrophy of the gastric fundus ducts, intestinal metaplasia or pseudopyloric metaplasia and dysplasia; thus, a combined determination of PG I and PG I/PG II ratio (PGR) may serve as a "serological biopsy" of the mucosa of the gastric fundus gland.

Pepsinogen is closely related to the occurrence of atrophic gastritis, which can cause the loss of the main cells of gastric mucosa, thereby affecting the secretory function thereof. Wherein, when the gastric mucosa is atrophied, the number of gastric mucosa main cells and glandular cells is reduced, thereby reducing the level of PG I in serum; since PG II can be secreted by cells of antrum and duodenal gland, PG II level can be in a relatively stable state. Therefore, if the atrophy degree of the gastric mucosa continues to progress and the range is expanded to affect the whole stomach, the content of PG I and PG II is reduced, and the reduction degree of PG I is more obvious than that of PG II, so that PGR is reduced. Di Mario et al found that low levels of PG I and PGR were significantly associated with atrophic gastritis in the stomach. The research of domestic scholars such as Chenjie also shows that the serum PG I level and PGR value of patients with atrophic gastritis are obviously reduced and are in negative correlation with the pathological change degree. Therefore, the reduction of the PG I and PGR levels in the serum can be used as an important serological index for judging the atrophy of the gastric mucosa. The statistical analysis of the R DEG C curve shows that the specificity of the PG I level is higher and can reach 80 percent when the atrophic gastritis is diagnosed, the sensitivity of the PGR ratio is higher and can reach 75 percent, and the diagnostic value is higher.

The prior art mainly comprises an immunoturbidimetry method, a chemiluminescence method, an enzyme-linked immunosorbent assay, an immunochromatography method, a latex-enhanced immunoturbidimetry method and the like. The latex enhanced immunoturbidimetry (LETIA) is simple and time-saving to operate, but needs to be completed by a full-automatic large-scale biochemical analyzer; the enzyme-linked immunosorbent assay (ELISA) has high sensitivity, large sample amount and quantitative detection, but long operation time and low automation; the Immunoturbidimetry (TIA) and the Chemiluminescence (CL) are sensitive and accurate, can be applied to a full-automatic biochemical analyzer, but require instruments and consume a long time, are suitable for processing a large number of samples, and cannot meet the aim of rapid detection.

Time resolved fluorescence analysis (TRFIA) is a relatively new type of detection means proposed in the 80's of the last century. The time-resolved fluorescence immunoassay is an emerging immunoassay technology which is established by combining a chromatography technology, an antigen-antibody specific immunoreaction and a fluorescence labeling technology. At present, in the time-resolved fluorescence analysis method, many rare earth nano materials applied to a time-resolved fluorescence probe have been reported, but a great space for improvement still exists. For example, the time-resolved fluorescent microspheres synthesized by swelling with rare earth complexes have the defects that the rare earth complexes are easy to leak and photo-bleach and the like; although the dissociation enhancement method (DELFLA) has high sensitivity, the fluorescence can be detected only by adding enhancement liquid; some fluorescent microspheres have the defects of weak fluorescence intensity and the like.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a rare earth detection test strip, a detection card, a kit and a preparation method, which can be used for rapidly detecting atrophic gastritis.

The technical scheme for realizing the purpose of the invention is as follows: a rare earth detection test strip for atrophic gastritis comprises a bottom liner, and a sample pad, a combination pad, a nitrocellulose membrane and absorbent paper which are sequentially lapped on the bottom liner along the length direction; rare earth nanometer fluorescent microspheres marked with monoclonal antibodies PG I and PG II and rabbit IgG antibodies are sprayed on the combination pads; sequentially scribing the nitrocellulose membrane through monoclonal antibodies PG I, PG II and corresponding monoclonal antibodies of goat anti-rabbit antibodies, and respectively taking the monoclonal antibodies as a first detection line, a second detection line and a quality control line; the first detection line is close to the combination pad, and the quality control line is close to the absorbent paper.

Further, the rare earth nano fluorescent microsphere is europium-doped gadolinium lutetium fluoride and has a chemical formula of NaGd _x Lu _1-y-x F ₄ yEu, wherein NaGd _x Lu _1-y-x F ₄ The doped ion is europium, and x and y are the doping mole percentages of the rare earth ions. x is in the range of 20% to 70%, preferably 40%, y is in the range of 0% to 30%, preferably 20%, and the particle size is 50nm to 150 nm.

Further, the preparation method of the rare earth nano fluorescent microsphere comprises the following steps:

step one, adding 3-6mL of oleic acid and 7-14mL of 1-octadecene into a 100mL three-neck round-bottom flask, and then adding x parts of Gd nitrate, Gd acetate or Gd chloride, y parts of Eu nitrate, Eu acetate or Eu chloride, and 1-x-y parts of Lu nitrate, Lu acetate or Lu chloride according to molar ratio, wherein x and y are rare earth ion doping mole percentage, the range of x is 20-70%, and the range of y is 0-30%;

step two, mixing and stirring at room temperature, vacuumizing, heating to 120 ℃, reacting for 20 minutes, heating to 160 ℃, and reacting for 10 minutes to obtain a transparent solution;

naturally cooling to 50 ℃, releasing vacuum, adding a mixed solution of 1-2mmol of NaOH and 1.6-3.4mmol of ammonium fluoride methanol, and reacting for 30 min;

heating to 100 ℃, exhausting air and ventilating for 3-4 times, introducing nitrogen, heating to 300 ℃, reacting for 1-2 hours, centrifuging at 6000rpm, washing for 3-4 times by using a cyclohexane-ethanol mixed solution, and dispersing in cyclohexane;

step five, transferring the probe to a water phase by using an acid washing method, and modifying carboxyl on the surface of the probe to obtain water-soluble NaGd with good dispersibility _x Lu _1-y-x F ₄ yEu rare earth nanometer fluorescent microsphere.

Furthermore, the rare earth nano fluorescent microsphere is stable under a ground state, and emits fluorescence with the wavelength range of 600nm to 620nm under the action of an excitation light source of 250nm to 380 nm.

Furthermore, the particle size of the rare earth nanometer fluorescent microsphere is 50 nm-150 nm.

Further, the size of the bottom lining is 80nm multiplied by 4mm, the size of the sample pad is 18nm multiplied by 4mm, the size of the combination pad is 15nm multiplied by 4mm, the size of the nitrocellulose membrane is 25nm multiplied by 4mm, and the size of the absorbent paper is 28nm multiplied by 4 mm.

The invention also provides a atrophic gastritis rare earth detection card, which comprises a card shell and a test strip arranged in the card shell; the test strip adopts the atrophic gastritis rare earth detection test strip; the card shell is provided with a sample adding hole and an observation window, the sample adding hole corresponds to a sample pad of the test strip, and the observation window corresponds to a marking position on the nitrocellulose membrane.

Furthermore, the clamping shell comprises an upper plastic shell and a lower plastic shell which are clamped with each other; the sampling hole and the observation window are arranged on the plastic upper shell.

The invention also provides a kit for detecting the atrophic gastritis by using the rare earth, which comprises a detection card, sample diluent and an ID card containing a calibration curve; the detection card adopts the atrophic gastritis rare earth detection card.

The preparation method of the atrophic gastritis rare earth detection kit is characterized by comprising the following steps:

step one, preparing rare earth nano fluorescent microspheres;

step two, activating the rare earth nano fluorescent microspheres;

marking monoclonal antibodies and rabbit IgG corresponding to PG I and PG II by rare earth nano fluorescent microspheres;

step four, preparing a sample pad;

step five, preparing a coating film;

and step six, sequentially and mutually bonding a sample pad, a bonding pad, a coating film and absorbent paper on the bottom lining to obtain a test paper board, and then cutting the test paper board into test paper strips.

Assembling the test strip in a card shell;

step eight, preparing a sample diluent;

and step nine, preparing the ID card containing the standard curve.

By adopting the technical scheme, the invention has the following beneficial effects: the invention utilizes the rare earth nanometer fluorescent microsphere detection technology, can avoid the fluorescence interference of a sample, the fluorescent microsphere is a rare earth fluoride luminescent nanometer material and has the advantages of low background, long luminescent life, strong fluorescent signal, high signal to noise ratio and the like, the microsphere is connected with the antibody by the covalent bond through the marker, the marker product is stable, and the invention has the characteristics of wide detection range, high sensitivity, high accuracy, quick, simple and convenient detection and the like.

Drawings

In order that the manner in which the above recited features of the present invention can be understood in detail, a more particular description of the invention, briefly summarized above, may be had by reference to embodiments, some of which are illustrated in the appended drawings

Fig. 1 is a schematic structural diagram of the test strip of the present invention.

FIG. 2 is a graph of a standard chart of PG I of the kit of example 1 of the present invention;

FIG. 3 is a graph of the standard PG II of the kit of example 1 of the present invention;

FIG. 4 is a graph showing the correlation between the results of the immunoassay of the kit of example 1 and the detection result of the detection of PG I by the Abbott chemiluminescence microparticle immunoassay.

FIG. 5 is a comparison of the correlation between the results of the detection by the kit of example 1 and the detection by the Yapei chemiluminescence microparticle immunoassay kit PGII.

The reference numbers in the drawings are:

the detection device comprises a sample pad 1, a combination pad 2, a nitrocellulose membrane 3, a first detection line 31, a second detection line 32, a quality control line 33 and absorbent paper 4.

Detailed Description

(example 1)

The kit for detecting the rare earth for atrophic gastritis comprises a detection card, a sample diluent and an ID card containing a calibration curve.

The detection card comprises a card shell and a test strip arranged in the card shell. Referring to fig. 1, the test strip comprises a bottom lining, and a sample pad 1, a binding pad 2, a nitrocellulose membrane 3 and absorbent paper 4 which are sequentially overlapped on the bottom lining along the length direction; rare earth nano fluorescent microspheres marked with PG I, PG II and rabbit IgG antibodies are sprayed on the binding pad 2; the nitrocellulose membrane is sequentially marked by monoclonal antibodies corresponding to PG I, PG II and rabbit IgG antibodies and respectively used as a first detection line 31, a second detection line 32 and a quality control line 33, wherein the first detection line 31 is close to the combination pad 2, and the quality control line 33 is close to the absorbent paper 4.

The size of the bottom lining is 80nm multiplied by 4mm, the size of the sample pad 1 is 18nm multiplied by 4mm, the size of the combination pad 2 is 15nm multiplied by 4mm, the size of the nitrocellulose membrane 3 is 25nm multiplied by 4mm, and the size of the absorbent paper 4 is 28nm multiplied by 4 mm.

The rare earth nano fluorescent microsphere is europium-doped gadolinium lutetium fluoride and has a chemical formula of NaGd _x Lu _1-y-x F ₄ yEu, wherein NaGd _x Lu _1-y-x F ₄ The doped ion is europium, and x and y are the doping mole percentages of the rare earth ions. x is in the range of 20% to 70%, preferably 40%, y is in the range of 0% to 30%, preferably 20%, and the particle size is 50nm to 150 nm. The rare earth nano fluorescent microsphere is stable under a ground state, and emits fluorescence with the wavelength range of 600nm to 620nm under the action of an excitation light source with the wavelength range of 250nm to 380 nm.

The clamping shell comprises an upper plastic shell and a lower plastic shell which are clamped with each other; the plastic upper shell is provided with a sample adding hole and an observation window, the position of the sample adding hole corresponds to the sample pad 1 of the test strip, and the position of the observation window corresponds to the marking position on the nitrocellulose membrane.

With NaGd _0。4 Lu0.4F ₄ 0.2Eu rare earth nano fluorescent microsphere as an example, the preparation method of the kit for detecting atrophic gastritis comprises the following steps:

step one, preparing NaGd ₀ 。4Lu0.4F ₄ 0.2Eu rare earth nano fluorescent microsphere, comprising the following steps:

s1.1, adding 4.5mL of oleic acid and 12.5mL of 1-octadecene into a 100mL three-neck round-bottom flask, and adding 1.6mmol of gadolinium acetate, 1.6mmol of lutetium acetate and 0.8mmol of europium acetate according to the mol ratio;

s1.2, mixing and stirring at room temperature, vacuumizing, heating to 120 ℃, reacting for 20 minutes, heating to 160 ℃, and reacting for 10 minutes to obtain a transparent solution;

s1.3, naturally cooling to 50 ℃, releasing vacuum, adding a mixed solution of 1mmol of NaOH and 1.6mmol of ammonium fluoride methanol, and reacting for 30 min;

s1.4, heating to 100 ℃, exhausting and ventilating for 3 times, introducing nitrogen, heating to 300 ℃, reacting for 1.5 hours, centrifuging at 6000rpm, washing for 3 times by using a cyclohexane-ethanol mixed solution, and dispersing in cyclohexane;

s1.5, transferring the probe to a water phase by using an acid washing method, and modifying carboxyl on the surface of the probe to obtain water-soluble NaGd with good dispersibility ₀ 。4Lu0.4F ₄ 0.2Eu rare earth nano fluorescent microsphere. The rare earth nanometer fluorescent microsphere has the diameter of 100nm, uniform appearance and good luminous performance.

And step two, activating the rare earth nano fluorescent microspheres. The method specifically comprises the following steps: after the rare earth nano fluorescent microspheres are treated by ultrasonic waves for 2min, 200 mul of rare earth nano fluorescent microspheres are taken and centrifuged at 14000rpm for 15min at a high speed, and precipitates are washed to 1ml by MES solution with the concentration of 100mM and the pH value of 6.0 and treated by ultrasonic waves for 2 min; adding 50 μ l of 100mg/ml carbodiimide, mixing for 5min, adding 100 μ l of 100mg/ml N-hydroxy-thiosuccinimide, mixing for 15min, centrifuging at 14000rpm for 15min, and washing the precipitate with MES solution with pH of 6.0 to 1 ml.

And step three, preparing the monoclonal antibodies and rabbit IgG corresponding to the rare earth nano fluorescent microspheres labeled PG I and PG II. The method specifically comprises the following steps: putting 200 mul of microspheres with solid content of 1% and particle size of 0.20 mu m into a round-bottom centrifuge tube, adding 100 mu g of PG I antibody, and marking; putting 200 mu L of microspheres with solid content of 1% and particle size of 0.20 mu m into a round-bottom centrifuge tube, adding 100 mu g of PG II antibody, and marking; putting 200 mu L of microspheres with solid content of 1% and particle size of 0.20 mu m into a round-bottom centrifuge tube, adding 120 mu g of rabbit IgG antibody, and marking; placing the mixture at room temperature in a dark place for coupling for 2h, sealing for 1h, washing with a storage solution for 2 times, centrifuging at 14000rpm for 15min, and obtaining precipitates, namely PG I-YG, PG II-YG and rabbit IgG-YG.

Step four, the sample pad 1 is prepared. The method specifically comprises the following steps: two lines of a sample pad 1 treatment solution (Tris-HCl containing 0.5% NaCl, 0.5% Tween, 20mM of 0.1% BSA and having a pH of 8.0) are uniformly sprayed in parallel on one side of the substrate close to the sample pad 1 in an amount of 4uL liquid/cm of the sample pad 1, and monoclonal antibodies PG I and PG II labeled with rare earth nano fluorescent microspheres and rabbit IgG antibodies are diluted in a microsphere diluent (20 mM Tris-HCl buffer containing 0.5% BSA and 25% sucrose) in an amount of 8%, 8% and 3% and then uniformly sprayed on one line (i.e. microsphere line) in an amount of 4uL liquid/cm of the sample pad 1. The mixture was placed in an oven and dried overnight at 37 ℃.

And step five, preparing a coating film. The method specifically comprises the following steps: respectively adjusting the concentrations of the monoclonal antibody PG I, the monoclonal antibody PG II and the goat anti-rabbit antibody to 1.0mg/mL, 1.0mg/mL and 0.8mg/mL of 1uL coating liquid amount/cm membrane by using coating buffer solutions (10 mM pH8.0PB buffer solution containing 0.85% NaCl and 0.05% ProClin 300), respectively serving as a first detection line 31, a second detection line 32 and a quality control line 33 to be parallelly scratched on a nitrocellulose membrane 3 for coating, respectively, enabling the quality control line 33 and the detection lines to be separated by 4mm, airing in an oven at the temperature of 37 ℃ for 10 hours at the humidity of less than 30%, and sealing and reserving.

And step six, adhering a sample pad 1 (with the size of 18mm multiplied by 300mm and made of glass fiber cotton), a bonding pad 2 (with the size of 15mm multiplied by 300mm and made of polyester film), a coating film (with the size of 25mm multiplied by 300mm and made of nitrocellulose) and absorbent paper 4 (with the size of 28mm multiplied by 300mm) to a bottom lining (with the size of 80mm multiplied by 300mm) in sequence in a mutually overlapped manner to obtain a test paper board, and cutting the test paper board into test paper strips with the width of 4mm according to requirements.

And step seven, assembling the test strip in a clamping shell formed by clamping and connecting the plastic upper shell and the plastic lower shell.

Step eight, preparing sample diluent which is Tris-hydrochloric acid buffer solution containing 1% Tween-20, 1% Tween-100 and 0.3% pH8.0, and subpackaging the sample diluent into centrifuge tubes by 180 ul/tube. The sample diluent is used for the chromatography sample.

And step nine, preparing an ID card (same batch of standard curves) containing a standard curve, measuring quality control products with different concentrations through a detection card, drawing the standard curve by taking the concentration of the quality control products as a horizontal coordinate and taking the ratio of fluorescence signals as a vertical coordinate, writing the standard curve into the ID card to generate a two-dimensional code, reading corresponding two-dimensional code information on the reagent card through a dry fluorescence immunoassay analyzer, and measuring the corresponding concentration.

The standard curve is drawn in the following process: adding PG I antigen quality control products (200ng/mL, 150ng/mL, 100ng/mL, 70ng/mL, 20ng/mL, 10ng/mL, 2 ng/mL) with different concentrations and PG II antigen quality control products (100ng/mL, 70ng/mL, 35ng/mL, 10ng/mL, 7ng/mL, 3ng/mL, 1ng/mL) into a detection card for 7 concentrations, setting three times for each concentration, respectively diluting PG I and PG II antigens by calf serum to obtain 1000ng/mL quality control product mother liquor, then diluting by calf serum to different quality control product concentrations, adding a sample diluent, carrying out chromatography for 15min, c, T line fluorescence signals and C/T values were read by AFS-1000 dry fluoroimmunoassay analyzer manufactured by Guangzhou Bluebo Biotech, Inc., and the experimental results and analysis are shown in the following table:

then, the concentrations of the PG I and PG II quality control substances and the average values of the sample signals T1/C and T2/C are respectively plotted as standard curves, the data of the curves are shown in the table, and the standard curves are shown in FIG. 4. Where the PG IR 1 value (correlation) was 0.9966 and PG IR R2 value was 0.9954, the concentrations of PG I and PG II contained in the samples were quantified by this scale.

The performance test of the test card of the present embodiment is as follows:

minimum detection limit: and (3) repeatedly measuring the zero-value sample for 20 times, calculating the mean M and standard deviation SD of 20 times of results, reporting the detection limit (M +2SD) of the method by adding two times of standard deviation to the blank mean, wherein the PG I result is 0.92ng/mL and meets the sensitivity of 1.0 ng/mL. PG II results were 0.86ng/mL, consistent with a sensitivity of 1.0 ng/mL. Linear range: PG I takes 7 concentration values between 2.0 and 200.0ng/mL, each concentration is repeatedly measured three times, the average value of the measured concentration and the theoretical concentration are subjected to linear analysis, a linear equation y is 0.961x +1.2197, r is 0.9986, and the kit has good correlation in the linear range of 2.0 to 200.0 ng/mL. PG II takes 7 concentration values between 1.0 and 100.0ng/mL, each concentration is repeatedly measured three times, the average value of the measured concentration and the theoretical concentration are subjected to linear analysis to obtain a linear equation y which is 1.0251x +0.0518, and r which is 0.9971, and the kit has good correlation in the linear range of 1.0 to 100.0 ng/mL.

Precision: taking three batches of the time-resolved fluorescence quantitative detection PG I/PG II kit prepared in the example 1, detecting 70 and 20ng/mL of repetitive quality control substances respectively by using the PG I quality control substances, detecting 10 times by using the repetitive quality control substances in each batch of the kit in parallel, wherein the CV of 70ng/mL in the three batches is respectively 4.03%, 6.01% and 4.93%, the CV of 20ng/mL in the three batches is respectively 5.14%, 6.39%, 7.83% and 5.54%, and the CV of 6.42% in the three batches is within 10%. 35ng/mL of repetitive quality control materials are respectively detected by the PG II quality control material, each batch of the kit is parallelly detected by the repetitive quality control materials for 10 times, the CV in three batches with the concentration of 35ng/mL is respectively 5.45%, 6.01% and 6.98%, the CV among the three batches is 5.98%, the CV in three batches with the concentration of 7ng/mL is respectively 3.69%, 8.38% and 8.96%, and the CV among the three batches is 7.36%, and the CV among the three batches is within 10%.

Accuracy: PG I selects a quality control substance with the concentration of 10ng/mL as a detection sample, the detection sample is divided into 3 parts with the same volume, 150ng/mL and 70ng/mL accuracy quality control substances are respectively added into 2 parts of the sample to prepare 2 recovered samples with different addition concentrations, and the concentration of the added substance to be detected is calculated. The same amount of the analyte-free solution was added to another sample to prepare a base sample. The samples were analyzed for 3 replicates and the mean was calculated. The recovery rate was calculated as (concentration of analytical sample-concentration of base sample)/concentration of addition x 100%. The recovery rate of the recovered sample is 99.65% at 150ng/mL, 102.28% at 70ng/mL and 100.96% on average. The deviation is within 10%. PG II selects a quality control substance with the concentration of 5ng/mL as a detection sample, the detection sample is divided into 3 parts with the same volume, 50ng/mL and 20g/mL accuracy quality control substances are respectively added into 2 parts of the sample, 2 recovery samples with different adding concentrations are prepared, and the concentration of the added substance to be detected is calculated. The recovery rate of 50ng/mL of the recovered sample was 102.55%, the recovery rate of 20ng/mL was 106.36%, and the average recovery rate was 104.46%. The deviation is within 10%.

The test results of the clinical samples of the kit of the present embodiment are as follows:

200 parts of plasma samples for detecting PG I and PG II in hospitals are respectively collected, and the detection and comparison are carried out by using the kit of the invention and a reagent kit for detecting PG I and PG II by an Yapei chemiluminescence micro-particle immunoassay method. In the kit, 20uL of a plasma sample is added into a sample diluent, 80uL of the plasma sample is added into a sample adding hole of a detection card after being uniformly mixed, the concentration is read by an AFS-1000 dry type fluorescence immunoassay analyzer produced by Guangzhou blue-Bo biological technology company Limited after chromatography is carried out for 15min, and the concentration detection is carried out on the same sample by respectively adopting a comparison system Yapei chemiluminescence micro-particle immunoassay method detection kit PG I and PG II. The linear analysis of the detection results of the two methods is shown in fig. 5, the correlation is good, the average relative deviation of PG IR is less than 10%, PG IIr is 0.9856, P is less than 0.05, and the average relative deviation is less than 10%, and the result meets the clinical analysis requirements and is suitable for clinical detection.

The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are only exemplary embodiments of the present invention, and are not intended to limit the present invention, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A rare earth test strip for atrophic gastritis is characterized in that:

comprises a bottom lining, a sample pad, a combination pad, a nitrocellulose membrane and absorbent paper, wherein the sample pad, the combination pad, the nitrocellulose membrane and the absorbent paper are sequentially lapped on the bottom lining along the length direction; rare earth nanometer fluorescent microspheres marked with monoclonal antibodies PG I and PG II and rabbit IgG antibodies are sprayed on the combination pads; the nitrocellulose membrane is sequentially scribed through monoclonal antibodies corresponding to the monoclonal antibody PG I, the monoclonal antibody PG II and the rabbit IgG antibody, and the scribed lines are respectively used as a first detection line, a second detection line and a quality control line; the first detection line is close to the combination pad, and the quality control line is close to the absorbent paper;

the rare earth nano fluorescent microsphere is europium-doped gadolinium lutetium fluoride and has a chemical formula of Na Gd _x Lu _1-y-x F ₄ yEu, wherein Na Gd _x Lu _1-y-x F ₄ Is taken as a substrate, the doping ions are Eu, x and y are the doping mole percentage of rare earth ions, x is 40 percent, y is 20 percent, and the grain diameter is 50nm to 150 nm;

the rare earth nano fluorescent microsphere is stable under a ground state, and emits fluorescence with the wavelength range of 600nm to 620nm under the action of an excitation light source with the wavelength range of 250nm to 380 nm.

2. A atrophic gastritis rare earth detection card is characterized in that:

comprises a card shell and a test strip arranged in the card shell; the test strip adopts the atrophic gastritis rare earth detection test strip of claim 1; the card shell is provided with a sample adding hole and an observation window, the sample adding hole corresponds to a sample pad of the test strip, and the observation window corresponds to a marking position on the nitrocellulose membrane.

3. The atrophic gastritis rare earth test card of claim 2, wherein:

the clamping shell comprises an upper plastic shell and a lower plastic shell which are clamped with each other; the sampling hole and the observation window are arranged on the plastic upper shell.

4. A rare earth detection kit for atrophic gastritis is characterized in that:

comprises a detection card, a sample diluent and an ID card containing a calibration curve; the detection card adopts the atrophic gastritis rare earth detection card of claim 3.

5. The method for preparing a rare earth detection kit for atrophic gastritis according to claim 4,

the method comprises the following steps: