1. Product name: Generic name: C-reactive protein (CRP) rapid quantification kit (colloidal gold method). Product name: CRP-DOT.
English name: Rapid quantitative Determination for C-Reactive Protein (CRP) by DIGFA method
2. Packaging specifications: 20 servings / box.
3. Intended use: This product can quickly and quantitatively determine CRP in serum or plasma on the Uppergold U2 gold standard method quantitative reader, which belongs to the category of POCT (Instant Inspection). Quantitative results can be obtained within 5 minutes. The quantitative range is 5-160 mg CRP / L. It is suitable for outpatient laboratories and other places where rapid results are required. At the same time of diagnosis of bacterial infection, it can be interpreted whether there is active disease . The quantitative determination of CRP with serum CRP 10mg / L as the judgment value (Cut Off) is used as an important indicator of bacterial infections and active diseases (serum and plasma CRP equivalent) at home and abroad. Other quantitative methods include particles with better accuracy and using more expensive instruments to facilitate immunoassays, radioimmunoassays, etc. They are more suitable for use when processing a large number of samples in a central laboratory.
4. Test principle: The reaction principle is the solid phase double antibody sandwich immunoassay. The sample to be tested is diluted and mixed with the CRP monoclonal antibody red colloidal gold conjugate in the reaction tube. The CRP in the sample specifically binds to the colloidal gold conjugate, and then the mixture is added and flows through the nitrate in the reaction plate of this kit The cellulose-based membrane, which contains the CRP bound to the red conjugate, can be specifically captured by the CRP antibody in the solid phase on the membrane, showing red spots. The red intensity of the spots can be measured with a U2 gold standard quantitative reader , Which is proportional to the CRP concentration in the sample. The CRP standard of this kit starts from CRM 470 (IFCC / BCR / CAP) international standard CRP.
V. Main components
Name of components and test equipment
Main active ingredients and concentration
Quantity
1. CRP reaction plate
CRP antibody, sensitization concentration 0.5-2.0mg / ml
20 pieces
2. CRP blocking solution
1% bovine serum albumin (BSA), pH7.4 PBS solution
1 × 3.0ml
3. CRP washing solution
pH7.4 PBS
1 × 7.0ml
4. CRP gold standard solution lyophilized product
CRP monoclonal antibody colloidal gold conjugate, 1% BSA
2 × 1.1ml
5. CRP gold standard lyophilized product complex solution
Double distilled water
1 × 4 ml
6. CRP sample diluent
0.1mol / L Glycine-NaOH buffer pH7.6, 0.03% Tween
20 tubes, volume injection bottle stickers
7. Reaction tube
Clean empty tube for single use
20
8. Instructions
1 serving
6. Storage conditions and expiration date: The kit is stored at 2 ~ 8 ℃, the shelf life is 12 months, and the gold standard solution will be used within one month after reconstitution. It should be protected from high temperature and sunlight during transportation, and refrigerated when the temperature is high. See packaging for production date.
7. Applicable instruments: Only applicable to the Uppergold U2, a gold standard spectrometer reader produced by Shanghai Aopu Biomedicine Co., Ltd.
8. Sample requirements: fresh serum or plasma, if there is obvious precipitation after thawing at low temperature, take the supernatant for centrifugation to determine (see notes for whole blood samples).
Nine, inspection method:
1. Remove the kit from the refrigerator and equilibrate at room temperature for at least half an hour. In the CRP gold standard solution freeze-dried product, accurately add the CRP freeze-dried product complex solution 1.1ml and shake well.
2. Use a clean tip to absorb 100 μl of the reconstituted CRP gold standard solution and add it to the empty reaction tube. The tip only aspirates the CRP gold standard solution to prevent any contamination.
3. Place the CRP reaction plate flat on the experimental table, and drop 2 drops of CRP blocking solution into the reaction well until it is completely infiltrated.
4. Fresh serum or plasma samples do not need pretreatment, frozen or stored at 4 ~ 8 ℃ for many days, centrifuge the sample and use the supernatant; accurately draw 10μl of serum or plasma into the CRP dilution tube (see bottle stickers for dilution volume, dilution factor Generally not less than 1: 160), mix well.
5. Pipette 100μl of the diluted sample into the 100μl CRP gold standard solution in the reaction tube, and mix well (the pipette must be mixed repeatedly for 10 times), and immediately add all the mixture to the well of the closed CRP reaction plate .
6. After complete infiltration, add 4 drops of CRP washing solution.
7. After the washing solution is completely infiltrated, read the CRP value within 5 minutes under the URP gold standard quantifier CRP serum / plasma item.
8. The Upper Gold U-2 quantitative reading instrument operates according to the instrument manual.
10. Reference value (reference range): 99% of healthy people have serum CRP below 10 mg / L, suggesting that the reference range for infection is ≥10 mg CRP / L.
11. Interpretation of test results: CRP is a human acute phase protein, a normal component that exists at low levels in the blood. When the human body has bacterial infections, active lesions, and large-scale tissue damage, it will increase rapidly. Return to the normal level.
In acute infections and large areas of trauma, serum CRP rises sharply. Serum CRP ≥ 10 mg CRP / L indicates infection. Mild and moderate bacterial infections usually only increase at low or moderate levels, and more than 50 mg CRP / L indicates extensive Sexual infection. Healthy people with a serum CRP value of 5-10 mg / L should be vigilant, and it is recommended to do ultra-micro CRP (hsCRP) when possible.
12. Limitations of the test method: The rheumatoid factor (RF), which is most likely to interfere with the double antibody sandwich method, has no effect on this determination. Severe chyle blood samples may block the pores of the nitrocellulose membrane, resulting in increased results due to reddish background. This method should not be used for determination.
13. Product Performance Index:
1. Specificity: The kit uses a specific CRP monoclonal antibody to label the analyte, and no other blood components have been detected in this CRP gold standard quantitative detection.
2 Precision: Repeat the test at a medically determined level (10mg / L CRP standard or serum sample), CV should be ≤20% (operated by skilled professionals, most cases CV≤15%)
3. Measurement range: The quantitative value measured by the instrument is 5-160mg CRP / L, and the reading interval is 1mg CRP / L.
4. Minimum detection limit: 5mg CRP / L
14. Matters needing attention
1. Since the dilution of the sample in this measurement is above 1: 160 (although the volume of the diluent occasionally varies slightly due to different batches of immunoreagent activity, the volume is noted on the bottle sticker, but it has no effect on the user), all Blood (including finger capillary blood) can also be used for sample determination, because the basis for judging the patient ’s bacterial infection or active disease is serum CRP, and the whole blood sample can be automatically converted to its corresponding serum CRP, plasma in U2 whole blood mode CRP is equivalent to serum CRP.
In pediatrics, it is difficult to collect venous blood, and it is easy to collect 10 μl of whole blood at the fingertip (the average proportion of blood cells in the whole blood sample is 40% of the total blood volume). In 6, the CRP washing solution was added in 3 times, 2 drops each time, after fully penetrating into the membrane, the next time the blood cell components can be washed away, and finally the converted serum CRP value was measured in the U2 CRP whole blood mode. If the patient has a routine blood test at the same time, please pay attention to the hematocrit measurement result (Hct%). If it deviates by more than 40%, you can manually multiply the corresponding correction factor in the following table to further correct it to make the whole blood converted CRP It is more consistent with its serum CRP value. Some imported kits still report the results as "CRP / whole blood" after such conversion, which is easy to misunderstand. It should be noted that the converted serum CRP value of whole blood is mg / L for doctors' reference.
Hct% value and correction factor
Hct%
Correction coefficient
Hct%
Correction coefficient
20-29
0.6
56-58
1.4
30-36
0.9
59-61
1.5
37-42
1.0
62-63
1.6
43-47
1.1
64-65
1.7
48-51
1.2
66-67
1.8
52-55
1.3
68-69
1.9
2. For in vitro diagnosis only, use within the validity period, do not mix reagent components in different batch kits.
3. Single-person operation is suitable, especially for those who are not familiar with the operation. Each step of the operation needs to be continuous. Once the sample or reagent is completely and completely infiltrated, the next step of the reagent should be added immediately, and it is not appropriate to pause.
4. The tip of the CRP colloidal gold must be clean, and the inside of the CRP colloidal gold bottle cap should not be contaminated. Otherwise, the CRP colloidal gold solution will self-coagulate after being contaminated. At this time, when the light is checked, the color of the CRP colloidal gold will be immediately after reconstitution The dark red turns to purple, the quality of CRP colloidal gold reagent is unqualified, which affects the quantitative results.
5. Serum and plasma stored at 4-8 ° C for several days should be centrifuged and the supernatant should be used to remove all fine precipitates in the sample to ensure that no particles block the pores in the cellulose membrane in the reaction plate to ensure white background. If the background is red, the quantitative result will be high, and the cause should be checked.
6. The reagent contains sodium azide as a preservative. Sodium azide is a toxic substance. Although its content does not exceed 0.15%, avoid splashing into the eyes and contact with the skin. Once it occurs, rinse immediately with tap water.
7. Body fluids such as serum, including healthy body fluids, are all potentially infectious substances. Operators should wear gloves. After testing, any items that come into contact with serum should be disinfected before discarding.
R & D Systems offers a complete range of complete ELISA kits. To ensure high quality and repeatability, all kits have undergone extremely strict quality inspections. To detect intracellular and extracellular proteins, the kit uses a variety of detection methods, including colorimetry, fluorescence and chemiluminescence.
Specification: 48 times / 96 times Price: 1500/2500
96 times kit composition:
1 30 times concentrated washing solution 20ml × 1 bottle 7 stop solution 6ml × 1 bottle
2 Enzyme label reagent 6ml × 1 bottle 8 standard (pg / ml) 0.5ml × 1 bottle
3 Enzyme label coating plate 12 wells × 8 strips 9 Standard diluent 1.5ml × 1 bottle
4 Sample diluent 6ml × 1 bottle 10 Instructions 1 copy
5 Developer A solution 6ml × 1 bottle 11 sealing film 2 sheets
6 Developer B liquid 6ml × 1 / bottle 12 sealed bag 1
Composition of 48 kits:
1 20 times concentrated washing solution 20ml × 1 bottle 7 stop solution 3ml × 1 bottle
2 Enzyme label reagent 3ml × 1 bottle 8 standard (pg / ml) 0.5ml × 1 bottle
3 Enzyme label coating plate 12 well × 4 strips 9 Standard dilution 1.5ml × 1 bottle
4 Sample diluent 3ml × 1 bottle 10 Instructions 1 copy
5 Developer A solution 3ml × 1 bottle 11 sealing film 2 sheets
6 Developer B liquid 3ml × 1 / bottle 12 sealed bag 1
Storage conditions: 2-8 degrees
Specimens: including serum, plasma, urine, pleural and ascites fluid, cerebrospinal fluid, cell culture supernatant, tissue homogenate, etc. Provide free testing services.
Test results within one week
Specimen collection method:
1. Serum:
After the blood coagulates naturally at room temperature for 10-20 minutes, centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again.
2. Plasma:
EDTA, sodium citrate or heparin should be selected as the anticoagulant according to the requirements of the specimen, mixed for 10-20 minutes, and centrifuged for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again.
3. Urine:
Collect with sterile tubes. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again. Pleural and ascites, cerebrospinal fluid refer to this practice.
4. Cell culture supernatant:
When detecting secreted components, collect them with sterile tubes. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully.
5. Cultivation of cells When testing the contents of cells, dilute the cell suspension with PBS (PH7.2-7.4) to a cell concentration of about 1 million / ml. By repeatedly freezing and thawing or adding tissue protein extraction reagents, the cells are destroyed and the intracellular components are released. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again.
6. Organize the specimen After cutting the specimen, weigh it. Add a certain amount of PBS, PH7.4. Quickly freeze and save with liquid nitrogen for later use. After the specimen melts, it still maintains a temperature of 2-8 ° C. Add a certain amount of PBS (PH7.4), or tissue protein extraction reagent, and homogenize the specimen by hand or homogenizer. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. After aliquoting, a portion is to be tested, and the rest is frozen for future use.
The company has a complete range of kits, including humans, rats, mice, rabbits, sheep, horses, cattle, guinea pigs, monkeys, fish, plants, food, etc. Nearly 1,000 kinds of kits have been put on the market, and are widely used by researchers Use, favored by customers
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