CN112649609A - Urotrypsinogen-2 detection reagent tablet and application thereof in pancreatic disease diagnosis - Google Patents

Urotrypsinogen-2 detection reagent tablet and application thereof in pancreatic disease diagnosis Download PDF

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CN112649609A
CN112649609A CN202011312848.XA CN202011312848A CN112649609A CN 112649609 A CN112649609 A CN 112649609A CN 202011312848 A CN202011312848 A CN 202011312848A CN 112649609 A CN112649609 A CN 112649609A
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detection
card
reagent
sample
urotrypsinogen
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曾曜龙
易剑峰
蔡其浩
胡小雄
施清蓉
钱伟
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Jiangxi Yingda Biotechnology Co ltd
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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    • G01N33/533—Production of labelled immunochemicals with fluorescent label
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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    • G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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    • G01—MEASURING; TESTING
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    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
    • G01N2333/976—Trypsin; Chymotrypsin
    • G—PHYSICS
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    • G01N2800/00—Detection or diagnosis of diseases
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Abstract

The invention discloses a prourotrypsin-2 detection reagent sheet and application of pancreatic disease diagnosis thereof, and the kit comprises a detection reagent sheet and a kit loaded with the detection reagent sheet, wherein the detection reagent sheet comprises a reagent body, the reagent body consists of a detection card and an IC card, the detection card consists of a plastic support plate and a test strip, the test strip consists of a PVC rubber plate, a glass fiber membrane, a nitrocellulose membrane, a goat anti-mouse polyclonal antibody, a TG-2 monoclonal antibody 1 and a fluorescence labeled TG-2 monoclonal antibody 2, and the kit can be stored at room temperature. The invention carries out the auxiliary diagnosis of pancreatic diseases by quantitatively measuring the content of trypsinogen 2(TG-2) in urine.

Description

Urotrypsinogen-2 detection reagent tablet and application thereof in pancreatic disease diagnosis
Technical Field
The invention belongs to the technical field of pancreatic disease diagnosis, and particularly relates to a urotrypsinogen-2 detection reagent tablet and application thereof in pancreatic disease diagnosis.
Background
Under normal conditions, pancreatic juice contains inactive trypsinogen in its glandular tissues. Pancreatic juice flows into duodenum continuously through the sphincter of choledochododex along pancreatic duct, and because bile exists in duodenum and mucosa of duodenal wall secretes enterokinase, under the action of the bile and the enterokinase, pancreatin starts to be converted into digestive enzyme with strong activity. Pancreatitis can be caused if the outflow tract is blocked and the excretion is not smooth.
At present, the existing pancreatitis detection mainly uses urine as a method for detecting pancreatic diseases, and compares the results of urine carboxypeptidase activated peptide (CAPAP) and Amylase (AMS) to investigate the clinical significance of the carboxypeptidase to the early diagnosis of acute pancreatitis, the carboxypeptidase adopts an immunochromatography colloidal gold method, the amylase adopts a nitrobenzene maltotrioside method, 30 cases of acute pancreatitis groups have results, the positive rate of CAPAP detection is 73.3 percent, the positive rate of AMS detection is 33.3 percent, 95 cases of abdominal pain groups have 16.8 percent, the positive rate of AMS detection is 5.3 percent, 179 cases of contrast groups have 2.2 percent, the positive rate of CAPAP detection is 3.4 percent, the consistency of the two methods is 3.0 percent, 82.2 percent, 10.9 percent and 3.9 percent, the acute pancreatitis group has 8 positive cases and 6 negative cases at the same time, the data of the detection method needs two substances, and clinical diagnosis of pancreatitis can be compared only by percentage.
Disclosure of Invention
The invention provides a reagent tablet for detecting trypsinogen-2 in urine and application thereof in diagnosing pancreatic diseases, which carries out auxiliary diagnosis on pancreatic diseases by quantitatively determining the content of trypsinogen 2(TG-2) in urine.
The technical scheme of the invention is realized as follows:
the reagent sheet for detecting the prourotrypsin-2 comprises a detection reagent sheet and a kit loaded with the detection reagent sheet, and is characterized in that the detection reagent sheet comprises a reagent body, the reagent body consists of a detection card and an IC card, and the detection card consists of a plastic support plate and a test strip.
In the reagent sheet for detecting the prourotrypsin-2, the test strip consists of a PVC rubber plate, a glass fiber membrane, a nitrocellulose membrane, a goat anti-mouse polyclonal antibody, a TG-2 monoclonal antibody 1 and a fluorescently-labeled TG-2 monoclonal antibody 2.
In the reagent sheet for detecting the prourotrypsin-2, the kit is stored at the temperature of 4-30 ℃, and the effective period of the detection card aluminum foil bag after being unsealed is 1 hour.
In the reagent sheet for detecting the prourotrypsin-2, the PVC rubber plate is provided with a sample pad, a combination pad and a water absorption part, and the nitrocellulose membrane is positioned between the combination pad and the water absorption material.
In the reagent sheet for detecting the prourotrypsin-2, the sample pad is provided with the sample adding area, the nitrocellulose membrane is provided with a T line and a C line, and the distance between the T line and the combination pad is larger than the distance between the C line and the combination pad.
In the reagent sheet for detecting the trypsinogen-2 in urine, the reagent body is suitable for quantitatively determining the content of the trypsinogen 2 in the urine, and the detection range of the reagent body is 20-2000 ng/mL.
In the application of the reagent tablet for detecting the urotrypsinogen-2 in the diagnosis of pancreatic diseases, the reagent tablet is used by the following steps;
1) before detection, the detection card and the sample are restored to room temperature;
2) determining that the IC card is matched with the batch number of the kit, placing the IC card in an instrument card reading area, and reading information;
3) opening the inner package of the detection card, and taking out the detection card; sucking 100 mu L of sample, vertically dropping the sample to the sample adding position of the detection card, and starting timing;
4) after adding the sample, clicking on a screen of a fluorescence immunoassay analyzer to start testing, and reacting the detection card for 10 minutes at room temperature;
5) inserting the detection card into a test card slot of a fluorescence immunoassay analyzer, and automatically testing the detection card by the analyzer;
6) finally, the detection result can be seen from the display screen of the fluorescence immunoassay analyzer, and the result can be printed by clicking the screen for printing.
The implementation of the reagent tablet for detecting the urotrypsinogen-2 and the application thereof in diagnosing the pancreatic diseases has the following beneficial effects: the scheme is provided with a detection reagent sheet and a kit loaded with the detection reagent sheet, wherein the reagent sheet is provided with a PVC (polyvinyl chloride) rubber plate, a glass fiber membrane, a nitrocellulose membrane, a goat anti-mouse polyclonal antibody, a TG-2 monoclonal antibody 1 and a fluorescence-labeled TG-2 monoclonal antibody 2, and when in detection, the urotrypsinogen-2 (TG-2) in a sample can be combined with the fluorescence-labeled urotrypsinogen-2 (TG-2) monoclonal antibody 2 on a fluorescence pad, and the free fluorescence-labeled urotrypsinogen-2 (TG-2) monoclonal antibody 2 is combined with the goat anti-mouse IgG polyclonal antibody at a quality control line to emit light, so that the auxiliary diagnosis of pancreatic diseases can be carried out by quantitatively determining the content of the trypsinogen 2(TG-2) in urine.
Drawings
FIG. 1 is a schematic flow chart showing the application of the reagent tablet for detecting urotrypsinogen-2 in the diagnosis of pancreatic diseases.
In the figure: the kit comprises a sample adding area 1, a combination pad 2, a T line 3, a C line 4, a nitrocellulose membrane 5, a PVC rubber plate 6, a water absorbing material 7, a sample 8 and a sample pad 9.
Detailed Description
The technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention.
Example 1
Property of reagent tablet for detecting urotrypsinogen-2
1. Product specification and specification of divisions thereof
1.1 specification: 1 part/box, 5 parts/box, 25 parts/box, 50 parts/box, 100 parts/box.
1.2 each portion test card is packaged in a separate aluminum foil bag.
2. Appearance character
2.1 the detection card is sealed with an aluminum foil bag without damage, tight sealing and air leakage.
2.2 the shell and the internal test paper of the detection card are neat and complete, and have no burr, no damage and no pollution.
2.3 Width of test strip
The width of the test strip is not less than 2.5 mm.
2.3.1 liquid migration velocity
The liquid moving speed is not lower than 10 mm/min.
2.4 accuracy
2.4.1 the relative deviation (B) should not exceed. + -. 15.0%.
2.4.2 the recovery rate should be in the range of 85.0-115.0%.
2.5 blank limit
The blank limit should not be higher than 20.0 ng/mL.
2.6 Linear Range
In the range of (20-2000) ng/mL of the kit, the correlation coefficient r should be not less than 0.9900.
2.7 repeatability
The Coefficient of Variation (CV) should be no more than 15.0%.
2.8 run-to-run differences
The relative range (R) should be not more than 15.0%
2.9 stability
Detecting in 2.3-2.7 with the kit stored until the effective period is exceeded, wherein the result meets the requirement of 2.3-2.7
3. The reagent body of the reagent tablet for detecting the trypsinogen-2 in urine comprises the components;
3.1 the reagent mainly comprises a detection card, an IC card and an instruction book. The detection card consists of a plastic support plate and a test strip, the test strip consists of a PVC rubber plate, a glass fiber membrane, a nitrocellulose membrane, a goat anti-mouse polyclonal antibody, a TG-2 monoclonal antibody 1, a fluorescently-labeled TG-2 monoclonal antibody 2 and the like, and the components of different batches cannot be mixed and exchanged.
4. The storage condition of the reagent box in the reagent tablet for detecting the trypsinogen-2 is determined;
the kit is stored at 4-30 ℃ and has a validity period of 18 months, the validity period of the test card aluminum foil bag after being unsealed is 1 hour, and the kit can be transported under the normal temperature condition.
5. The invention is applicable to instruments;
the device is suitable for an LD1800 dry type fluorescence immunoassay analyzer and an LD1800S dry type fluorescence immunoassay analyzer which are produced by the company; AFS-1000 dry fluoroimmunoassay analyzer, AFS2000A dry fluoroimmunoassay analyzer, and AFS3000B dry fluoroimmunoassay analyzer manufactured by Guangzhou Bluebo Biotechnology Ltd.
Requirement for 6 samples
6.1 for human urine samples, other body fluids and samples may not give accurate results.
6.2 samples were collected according to routine clinical medicine techniques without special treatment. If the detection cannot be carried out in time, the urine sample can be stored for 4 hours at room temperature (18-28 ℃) or 48 hours at 2-8 ℃.
6.3 samples must be returned to room temperature before testing and mixed until ready for use, suggesting a fresh sample.
6.4 No obvious haemolytic, jaundice and lipid turbidity samples were used. When the concentration of bilirubin in the sample is less than or equal to 200 mu mol/L, the concentration of hemoglobin is less than or equal to 4.0g/L, and the concentration of triglyceride is less than or equal to 10mmol/L, the determination result is not obviously influenced.
6.5. Samples above the upper limit of the linear range are not recommended for dilution.
(II) application step of urotrypsinogen-2 detection reagent tablet in pancreatic disease diagnosis
7 appearance Properties
Visual inspection under natural light with corrected vision should result in compliance with 2.1 requirements.
7.2 test strip Width
Randomly extracting a human test card, taking out the test paper in the card, measuring the width of the test paper by using a vernier caliper for 1 time, wherein the result meets the requirement of 2.2.
7.3 liquid migration speed
And (3) operating according to the specification, randomly drawing a person detection card, adding the sample liquid into the sample adding hole, starting timing by using a stopwatch when the liquid is dripped into the sample adding hole, stopping timing until the liquid runs out of the detection window, recording the time used as (t), measuring the moving length of the liquid by using a vernier caliper (not containing absorbent paper) as (L), calculating the L/t as the moving speed of the liquid, and meeting the requirement of 2.3 as a result.
7.4 accuracy
7.4.1 determination of national Standard substance (GBW) or other reference substances (RM) which can be used for evaluating conventional measurement procedures or enterprise reference substances or commercial quality control substances at high and low two-level concentrations, repeating the determination 3 times, and taking the mean value of the test results
Figure BDA0002790355180000051
The relative deviation (B) is calculated according to the formula (1), and the result meets the requirement of 2.4.1.
Figure BDA0002790355180000052
In the formula: b is relative deviation;
Figure BDA0002790355180000053
the mean value of the sample measurement results is obtained; TV is the reference target value.
7.5 blank limit
And (3) taking the reference substance matrix as a sample for detection, repeating the detection for 20 times, and calculating the mean value and the standard deviation of the detection results of 20 times, wherein the mean value plus 2 times of the standard deviation is a blank limit, and the result meets the requirement of 2.5.
7.6 Linear Range
The high value sample close to the upper limit of the linear range and the low value sample close to the lower limit of the linear range are mixed to 5 dilution concentrations (xi), the dilution concentrations are tested by a kit for at least 2 times, and the average value (yi) of the measurement results is obtained. The linear regression equation was calculated using the dilution concentration (xi) as an independent variable and the measurement result mean (yi) as a dependent variable. The correlation coefficient (r) of the linear regression is calculated according to equation (3), and the result should meet the requirement of 2.6.
Figure BDA0002790355180000061
7.7 repeatability
Detecting with reference sample or quality control material with high and low levels of concentration, repeating detection for 10 times at each level, and calculating average value of detection results
Figure BDA0002790355180000062
And Standard Deviation (SD), calculating the Coefficient of Variation (CV) of each level according to the formula (4), the formula (5) and the formula (6), and ensuring that the result meets the requirement of 2.7.
Figure BDA0002790355180000063
Figure BDA0002790355180000064
Figure BDA0002790355180000065
7.8 run-to-run differences
The reference samples or quality control products with two levels of concentration are respectively detected by 3 kits of different batches, each batch of each level is detected for 5 times, the average value i (i is 1,2 and 3) is respectively calculated, the relative range (R) of the measured average values of the 3 kits of batches is calculated according to the formulas (7) and (8), and the result meets the requirement of 2.8.
Figure BDA0002790355180000071
Figure BDA0002790355180000072
In the formula:
Figure BDA0002790355180000073
-mean of 3 reagent batches
Figure BDA0002790355180000074
——
Figure BDA0002790355180000075
Maximum value of
Figure BDA0002790355180000076
——
Figure BDA0002790355180000077
Minimum value of
7.9 stability
And (3) taking the kit stored until the kit exceeds the effective period, and detecting in the step 2.3-2.7 according to a method shown in the step 3.3-3.7, wherein the result meets the requirement of 2.9.
8.0 principle of examination
The kit adopts a double-antibody sandwich method, a detection line of a nitrocellulose membrane is coated with a urotrypsinogen-2 (TG-2) monoclonal antibody 1 from a mouse, a quality control line is coated with a goat anti-mouse IgG polyclonal antibody, and a fluorescent pad is fixed with a fluorescent label of the urotrypsinogen-2 (TG-2) monoclonal antibody 2 from the mouse. During detection, the urotrypsinogen-2 (TG-2) in a sample can be combined with a fluorescence-labeled urotrypsinogen-2 (TG-2) monoclonal antibody 2 on a fluorescence pad to form an immune complex, the immune complex moves along a membrane under the action of chromatography, the immune complex is combined with a coated urotrypsinogen-2 (TG-2) monoclonal antibody 1 to emit light when passing through a detection line, and the free fluorescence-labeled urotrypsinogen-2 (TG-2) monoclonal antibody 2 is combined with a goat anti-mouse IgG polyclonal antibody at a quality control line to emit light. The fluorescence signal is captured by the fluorescence immunoassay instrument, and the urotrypsin zymogen-2 (TG-2) in the sample is automatically converted and detected through signal conversion and the prepared standard curve.
9.0 test method
Before use, please carefully read the reagent instruction and the operation manual of the fluorescence immunoassay analyzer, and the related operations are performed strictly according to the reagent instruction, otherwise, reliable results cannot be guaranteed.
1. The test card and sample were returned to room temperature (15-30 deg.C) prior to testing.
2. And (4) determining that the IC card is matched with the batch number of the kit, placing the IC card in an instrument card reading area, and reading information.
3. Opening the inner package of the detection card, and taking out the detection card; sucking 100 mu L of sample, vertically dropping the sample to the sample adding position of the detection card, and starting timing;
4. after adding the sample, clicking a 'start test' on a screen of a fluorescence immunoassay analyzer, and reacting the detection card for 10 minutes at room temperature; inserting the detection card into a test card slot of a fluorescence immunoassay analyzer, and automatically testing the detection card by the analyzer; the detection result can be seen from the display screen of the fluorescence immunoassay analyzer, and the result can be printed by clicking the 'print' on the screen.
Example 2
Property of reagent tablet for detecting urotrypsinogen-2
1. Product specification and specification of divisions thereof
1.1 specification: 1 part/box, 5 parts/box, 25 parts/box, 50 parts/box, 100 parts/box.
1.2 each portion test card is packaged in a separate aluminum foil bag.
2. Appearance character
2.1 the detection card is sealed with an aluminum foil bag without damage, tight sealing and air leakage.
2.2 the shell and the internal test paper of the detection card are neat and complete, and have no burr, no damage and no pollution.
2.3 Width of test strip
The width of the test strip is not less than 2.5 mm.
2.3.1 liquid migration velocity
The liquid moving speed is not lower than 10 mm/min.
2.4 accuracy
2.4.1 the relative deviation (B) should not exceed. + -. 15.0%.
2.4.2 the recovery rate should be in the range of 85.0-115.0%.
2.5 blank limit
The blank limit should not be higher than 20.0 ng/mL.
2.6 Linear Range
In the range of (20-2000) ng/mL of the kit, the correlation coefficient r should be not less than 0.9900.
2.7 repeatability
The Coefficient of Variation (CV) should be no more than 15.0%.
2.8 run-to-run differences
The relative range (R) should be not more than 15.0%
2.9 stability
Detecting in 2.3-2.7 with the kit stored until the effective period is exceeded, wherein the result meets the requirement of 2.3-2.7
3. The reagent body of the reagent tablet for detecting the trypsinogen-2 in urine comprises the components;
3.1 the reagent mainly comprises a detection card, an IC card and an instruction book. The detection card consists of a plastic support plate and a test strip, the test strip consists of a PVC rubber plate, a glass fiber membrane, a nitrocellulose membrane, a goat anti-mouse polyclonal antibody, a TG-2 monoclonal antibody 1, a fluorescently-labeled TG-2 monoclonal antibody 2 and the like, and the components of different batches cannot be mixed and exchanged.
4. The storage condition of the reagent box in the reagent tablet for detecting the trypsinogen-2 is determined;
the kit is stored at 4-30 ℃ and has a validity period of 18 months, the validity period of the test card aluminum foil bag after being unsealed is 1 hour, and the kit can be transported under the normal temperature condition.
5. The invention is applicable to instruments;
the device is suitable for an LD1800 dry type fluorescence immunoassay analyzer and an LD1800S dry type fluorescence immunoassay analyzer which are produced by the company; AFS-1000 dry fluoroimmunoassay analyzer, AFS2000A dry fluoroimmunoassay analyzer, and AFS3000B dry fluoroimmunoassay analyzer manufactured by Guangzhou Bluebo Biotechnology Ltd.
Requirement for 6 samples
6.1 for human urine samples, other body fluids and samples may not give accurate results.
6.2 samples were collected according to routine clinical medicine techniques without special treatment. If the detection cannot be carried out in time, the urine sample can be stored for 4 hours at room temperature (18-28 ℃) or 48 hours at 2-8 ℃.
6.3 samples must be returned to room temperature before testing and mixed until ready for use, suggesting a fresh sample.
6.4 No obvious haemolytic, jaundice and lipid turbidity samples were used. When the concentration of bilirubin in the sample is less than or equal to 200 mu mol/L, the concentration of hemoglobin is less than or equal to 4.0g/L, and the concentration of triglyceride is less than or equal to 10mmol/L, the determination result is not obviously influenced.
6.5. Samples above the upper limit of the linear range are not recommended for dilution.
(II) application step of urotrypsinogen-2 detection reagent tablet in pancreatic disease diagnosis
7 appearance Properties
Visual inspection under natural light with corrected vision should result in compliance with 2.1 requirements.
7.2 test strip Width
Randomly extracting a human test card, taking out the test paper in the card, measuring the width of the test paper by using a vernier caliper for 1 time, wherein the result meets the requirement of 2.2.
7.3 liquid migration speed
And (3) operating according to the specification, randomly drawing a person detection card, adding the sample liquid into the sample adding hole, starting timing by using a stopwatch when the liquid is dripped into the sample adding hole, stopping timing until the liquid runs out of the detection window, recording the time used as (t), measuring the moving length of the liquid by using a vernier caliper (not containing absorbent paper) as (L), calculating the L/t as the moving speed of the liquid, and meeting the requirement of 2.3 as a result.
7.4 accuracy
7.4.1 adding a certain volume of standard solution or pure product into the human matrix sample for testing, repeatedly measuring each sample for 3 times, taking the average value of the test results, and calculating the recovery rate according to the formula (2), wherein the result meets the requirement of 2.4.2.
Figure BDA0002790355180000101
In the formula: r isRecovery rate; v is the volume of the standard solution added; v0Is the volume of the human sample; c, detecting the concentration of the human sample after the human sample is added into the standard solution; c. C0The detected concentration of the human-derived sample; c. CsIs the concentration of the standard solution;
7.5 blank limit
And (3) taking the reference substance matrix as a sample for detection, repeating the detection for 20 times, and calculating the mean value and the standard deviation of the detection results of 20 times, wherein the mean value plus 2 times of the standard deviation is a blank limit, and the result meets the requirement of 2.5.
7.6 Linear Range
The high value sample close to the upper limit of the linear range and the low value sample close to the lower limit of the linear range are mixed to 5 dilution concentrations (xi), the dilution concentrations are tested by a kit for at least 2 times, and the average value (yi) of the measurement results is obtained. The linear regression equation was calculated using the dilution concentration (xi) as an independent variable and the measurement result mean (yi) as a dependent variable. The correlation coefficient (r) of the linear regression is calculated according to equation (3), and the result should meet the requirement of 2.6.
Figure BDA0002790355180000111
7.7 repeatability
Detecting with reference sample or quality control material with high and low levels of concentration, repeating detection for 10 times at each level, and calculating average value of detection results
Figure BDA0002790355180000112
And Standard Deviation (SD), calculating the Coefficient of Variation (CV) of each level according to the formula (4), the formula (5) and the formula (6), and ensuring that the result meets the requirement of 2.7.
Figure BDA0002790355180000113
Figure BDA0002790355180000114
Figure BDA0002790355180000115
7.8 run-to-run differences
The reference samples or quality control products with two levels of concentration are respectively detected by 3 kits of different batches, each batch of each level is detected for 5 times, the average value i (i is 1,2 and 3) is respectively calculated, the relative range (R) of the measured average values of the 3 kits of batches is calculated according to the formulas (7) and (8), and the result meets the requirement of 2.8.
Figure BDA0002790355180000116
Figure BDA0002790355180000117
In the formula:
Figure BDA0002790355180000121
-mean of 3 reagent batches
Figure BDA0002790355180000126
——
Figure BDA0002790355180000123
Maximum value of
Figure BDA0002790355180000127
——
Figure BDA0002790355180000125
Minimum value of
7.9 stability
And (3) taking the kit stored until the kit exceeds the effective period, and detecting in the step 2.3-2.7 according to a method shown in the step 3.3-3.7, wherein the result meets the requirement of 2.9.
8.0 principle of examination
The kit adopts a double-antibody sandwich method, a detection line of a nitrocellulose membrane is coated with a urotrypsinogen-2 (TG-2) monoclonal antibody 1 from a mouse, a quality control line is coated with a goat anti-mouse IgG polyclonal antibody, and a fluorescent pad is fixed with a fluorescent label of the urotrypsinogen-2 (TG-2) monoclonal antibody 2 from the mouse. During detection, the urotrypsinogen-2 (TG-2) in a sample can be combined with a fluorescence-labeled urotrypsinogen-2 (TG-2) monoclonal antibody 2 on a fluorescence pad to form an immune complex, the immune complex moves along a membrane under the action of chromatography, the immune complex is combined with a coated urotrypsinogen-2 (TG-2) monoclonal antibody 1 to emit light when passing through a detection line, and the free fluorescence-labeled urotrypsinogen-2 (TG-2) monoclonal antibody 2 is combined with a goat anti-mouse IgG polyclonal antibody at a quality control line to emit light. The fluorescence signal is captured by the fluorescence immunoassay instrument, and the urotrypsin zymogen-2 (TG-2) in the sample is automatically converted and detected through signal conversion and the prepared standard curve.
9.0 test method
Before use, please carefully read the reagent instruction and the operation manual of the fluorescence immunoassay analyzer, and the related operations are performed strictly according to the reagent instruction, otherwise, reliable results cannot be guaranteed.
1. The test card and sample were returned to room temperature (15-30 deg.C) prior to testing.
2. And (4) determining that the IC card is matched with the batch number of the kit, placing the IC card in an instrument card reading area, and reading information.
3. Opening the inner package of the detection card, and taking out the detection card; sucking 100 mu L of sample, vertically dropping the sample to the sample adding position of the detection card, and starting timing;
4. after adding the sample, clicking a 'start test' on a screen of a fluorescence immunoassay analyzer, and reacting the detection card for 10 minutes at room temperature; inserting the detection card into a test card slot of a fluorescence immunoassay analyzer, and automatically testing the detection card by the analyzer; the detection result can be seen from the display screen of the fluorescence immunoassay analyzer, and the result can be printed by clicking the 'print' on the screen. Thus, the object of the present invention has been accomplished.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims (7)

1. The reagent sheet for detecting the prourotrypsin-2 comprises a detection reagent sheet and a kit loaded with the detection reagent sheet, and is characterized in that the detection reagent sheet comprises a reagent body, the reagent body consists of a detection card and an IC card, and the detection card consists of a plastic support plate and a test strip.
2. The urotrypsinogen-2 detection reagent sheet according to claim 1, wherein the test strip is composed of a PVC rubber plate, a glass fiber membrane, a nitrocellulose membrane, a goat anti-mouse polyclonal antibody, a TG-2 monoclonal antibody 1 and a fluorescently-labeled TG-2 monoclonal antibody 2.
3. The reagent sheet for detecting trypsinogen-2 according to claim 1, wherein the reagent kit is stored at 4-30 ℃ and the effective period of the detection card aluminum foil bag is 1 hour after the detection card aluminum foil bag is opened.
4. The urotrypsinogen-2 detection reagent sheet according to claim 2, wherein the PVC rubber plate is provided with a sample pad, a combination pad and a water absorption part, and the nitrocellulose membrane is positioned between the combination pad and the water absorption material.
5. The urotrypsinogen-2 detection reagent sheet according to claim 4, wherein the sample pad is provided with a sample application region, the nitrocellulose membrane is provided with a T line and a C line, and the distance between the T line and the binding pad is larger than the distance between the C line and the binding pad.
6. The urinary trypsinogen-2 detection reagent sheet according to claim 1, wherein the reagent body is suitable for quantitative determination of the content of trypsinogen 2 in urine, and the detection range of the reagent body is 20-2000 ng/mL.
7. The use of the urotrypsinogen-2 detection reagent tablet according to any one of claims 1-6 for the diagnosis of pancreatic diseases, wherein the reagent tablet is used in the following steps;
1) before detection, the detection card and the sample are restored to room temperature;
2) determining that the IC card is matched with the batch number of the kit, placing the IC card in an instrument card reading area, and reading information;
3) opening the inner package of the detection card, and taking out the detection card; sucking 100 mu L of sample, vertically dropping the sample to the sample adding position of the detection card, and starting timing;
4) after adding the sample, clicking on a screen of a fluorescence immunoassay analyzer to start testing, and reacting the detection card for 10 minutes at room temperature;
5) inserting the detection card into a test card slot of a fluorescence immunoassay analyzer, and automatically testing the detection card by the analyzer;
6) finally, the detection result can be seen from the display screen of the fluorescence immunoassay analyzer, and the result can be printed by clicking the screen for printing.
CN202011312848.XA 2020-11-20 2020-11-20 Urotrypsinogen-2 detection reagent tablet and application thereof in pancreatic disease diagnosis Pending CN112649609A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202011312848.XA CN112649609A (en) 2020-11-20 2020-11-20 Urotrypsinogen-2 detection reagent tablet and application thereof in pancreatic disease diagnosis

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202011312848.XA CN112649609A (en) 2020-11-20 2020-11-20 Urotrypsinogen-2 detection reagent tablet and application thereof in pancreatic disease diagnosis

Publications (1)

Publication Number Publication Date
CN112649609A true CN112649609A (en) 2021-04-13

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Application Number Title Priority Date Filing Date
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Country Status (1)

Country Link
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Application publication date: 20210413