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Antistreptolysin "O" latex nephelometric immunoassay detection kit and preparation method

A streptolysin and detection kit technology, applied in the biological field, can solve the problems of narrow linear range of reagents, low analytical sensitivity, stability, poor analytical sensitivity and linear range of reagents, etc., to ensure repeatability and accuracy, Wide linear range, fast detection effect

Inactive Publication Date: 2019-08-20
BIOSINO BIO TECH & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] CN201510227792.0 discloses a detection kit for antistreptolysin "O", which uses chemical crosslinking to crosslink hemolysin "O" with carboxylated polystyrene latex to form an ASO reagent. The linear range of the prepared reagent is narrow, and the analytical sensitivity is low, which is not conducive to clinical promotion and use
[0006] At present, domestic ASO detection mainly relies on imported reagents. There are relatively few domestic reagents on the market, and there are problems such as stability, analytical sensitivity, and poor linear range of reagents, which affect the accuracy of clinical test results. Imported reagents are relatively expensive. It is expensive and causes a heavy burden to patients, and there is an urgent need for a product that is easy to operate, high in sensitivity, good in accuracy, and low in cost

Method used

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  • Antistreptolysin "O" latex nephelometric immunoassay detection kit and preparation method
  • Antistreptolysin "O" latex nephelometric immunoassay detection kit and preparation method
  • Antistreptolysin "O" latex nephelometric immunoassay detection kit and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Preparation method of antistreptolysin "O" latex immunoturbidity detection kit

[0046] The kit provided in this example is based on the latex immunoturbidimetric method and includes liquid dual reagents (reagent R1 and reagent R2). Among them, reagent R1 is a diluent (reaction buffer), and reagent R2 is a solution containing ASO latex microspheres (antistreptolysin "O" latex reagent). ASO latex reagent was prepared by coupling hemolysin "O" with carboxypolystyrene latex microspheres with imine and N-hydroxysuccinimide. The preparation method of the kit is as follows:

[0047] 1. Solution preparation

[0048] Preparation of activation solution: Weigh 10.66g of activator MES into a beaker, add 1L of purified water, stir and mix well, adjust the pH to 6.1 with NaOH solution, and store at 4°C for later use.

[0049] Preparation of coupling solution PBS: weigh 2.865g of Na 2 HPO 4 12H 2 O in a beaker, weigh 0.272g of KH in turn 2 PO 4 ·H 2 O, 7.948g of Na...

Embodiment 2

[0057] Example 2 Application of Antistreptolysin "O" Detection Kit

[0058] 1. Applicable instruments: HITACHI 7180, OLYMPUS AU400 and TOSHIBA 40FR automatic biochemical analyzer.

[0059] 2. Samples to be tested: Serum collected by clinical standard methods can be stored stably for one week at 2-8°C.

[0060] 3. See Table 1 for the specific testing procedures.

[0061] Table 1

[0062]

[0063]

[0064] The components of the calibrator are as follows: a calibrator solution containing 1% BSA, 0.1% Proclin-300 and 500 IU / mL antistreptolysin "O" prepared with 50 mM PBS buffer as a solvent.

[0065] 4. Calculation result: ASO concentration in the sample (IU / mL) = ΔAT / ΔAS × calibration solution concentration

[0066] In the formula: ΔAT is the absorbance of the sample tube compared with the absorbance of the blank tube.

[0067] ΔAS is the absorbance of the calibration tube compared to the absorbance of the blank tube.

[0068] 5. Reference value range: ASO≤195IU / mL.

...

Embodiment 3

[0073] Example 3 Antistreptolysin "O" detection kit linear detection range test

[0074] Compare and dilute the ASO sample with a concentration of 1500IU / mL into a gradient concentration, use this kit to measure each concentration three times, take the average value, and calculate the linear correlation coefficient with the theoretical value. The test results are shown in Table 2, and the linear correlation sex as figure 1 shown.

[0075] Table 2 Detection results of ASO gradient concentration

[0076]

[0077] figure 1 The corresponding linear correlation equation is: y=1.0183x-4.3588, the correlation coefficient R 2 =0.9998, the results show that the linear detection range of this kit is 20-1500IU / mL, and the detection limit can reach 20IU / mL.

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Abstract

The invention provides an antistreptolysin "O" latex nephelometric immunoassay detection kit and a preparation method thereof. The kit is a liquid double reagent used on a biochemical analyzer; the reagent R1 is a buffer system with a 0.1% to 5% w / v sensitizer, a 0.01% to 0.5% w / v surfactant, a 0.5% to 5% w / v protectant and a 0.02% to 0.2% w / v preservative added; and the reagent R2 is an antistreptolysin "O" latex reagent. Compared with the commercially available ASO kit, the kit provided by the invention has the advantages of high precision, high sensitivity, wide linear range, high specificity and good accuracy, and provides an important basis for auxiliary diagnosis of clinical ASO related diseases.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an antistreptolysin "O" latex immunoturbidimetric detection kit and a preparation method. Background technique [0002] Streptolysin "O" is one of the metabolites of hemolytic streptococcus, a protein containing -SH group, which can dissolve red blood cells and is easily oxidized. It can temporarily lose the hemolytic ability and form -SS group in contact with air. However, it can be revived with the help of a reducing agent, and has hemolysis again, and hemolysin "O" has a strong antigenicity. After a person is infected by streptococcal, ASO (antistreptolysin "O") begins to increase 7-10 days after streptococcal infection, and reaches a peak 2-3 weeks after acute infection. The research results of Johnson et al. showed that the elevated ASO would last for a long time, and 83.3% of the subjects still had elevated ASO after 6 months. ASO positive is common in pharyngitis...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/544G01N33/543G01N33/531
CPCG01N33/68G01N33/544G01N33/54313G01N33/531G01N2333/47
Inventor 张勇
Owner BIOSINO BIO TECH & SCI