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Kit for detecting human parvovirus IgM antibody

A parvovirus and kit technology, which is applied in the field of kits for detecting human parvovirus IgM antibodies, can solve the problems of narrow linear range, inaccurate sample addition, and human errors in naked eye interpretation results, and achieve wide detection linear range and high detection results. Accurate, short detection time effect

Inactive Publication Date: 2019-03-08
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Antibody detection is currently the main method for clinical diagnosis and epidemiological investigation of B19 infection. The detection methods mainly include ELISA and colloidal gold. The shortcomings are as follows: 1. Manual operation, inaccurate sample addition; long operation time and slow process It is cumbersome and prone to operational errors; these errors and errors will affect the accuracy and precision of the test results
2. The detection process is in an open space, which may easily cause cross-contamination between various reagents, thereby affecting the accuracy of the detection results
3. In terms of methodology, the sensitivity of colloidal gold analysis is low, and there are human errors in the results of naked eye interpretation
4. All are qualitative products with a narrow linear range; qualitative products use a simple color reaction point to determine the result, which is prone to false positive or false negative results

Method used

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  • Kit for detecting human parvovirus IgM antibody
  • Kit for detecting human parvovirus IgM antibody
  • Kit for detecting human parvovirus IgM antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Preparation of a kit for detecting human parvovirus IgM antibody

[0017] 1. Preparation of B19-IgM magnetic particle suspension

[0018] 1.1 Take 30ul of the mixed carboxy surface magnetic bead stock solution and wash 5 times with 300ul of PBS buffer.

[0019] 1.2 Activate the magnetic particles with 20mg / ml EDC and 20mg / ml NHS for 1 hour.

[0020] 1.3 Wash the activated magnetic particles 3 times with pH 4.75 buffer B.

[0021] 1.4 Add mouse anti-human IgM μ chain monoclonal antibody according to the amount of 0.25ug / test, and coat for 2 hours.

[0022] 1.5 Seal 4 times with sealing solution.

[0023] 1.6 Add 3ml of sealing solution for preservation.

[0024] The sealing solution used is prepared by 0.01M PH7-8 PBS buffer solution, which contains 1‰ (v / v) preservative Proclin 300, 0.2‰ (w / v) preservative NaN 3 , 3% (w / v) stabilizer bovine serum albumin, 1‰ (v / v) surfactant Triton X-100, 5% (v / v) glycerin.

[0025] 2. Preparation of magnetic particle B1...

Embodiment 2

[0038] Embodiment 2 The usage method of kit of the present invention

[0039] 1. Sample requirements

[0040]1.1 Use correct medical technology to collect whole blood samples, centrifuge after the blood cells are coagulated at room temperature, and extract serum for testing. Plasma with EDTA (1.5g / L whole blood), sodium citrate (10.9mmol / L whole blood) or heparin sodium (0.1-0.2mg / mL whole blood) as anticoagulant can also be used for detection.

[0041] 1.2 Serum should not be left at room temperature for more than 8 hours after collection. If it is not tested within 8 hours, the sample should be placed in a refrigerator at 2-8°C; if it needs to be stored or transported for a long time, it should be frozen at below -20°C to avoid repeated freezing and thawing. Return to room temperature before use, shake gently to mix.

[0042] 2. Inspection method

[0043] 2.1 Consumables inspection: load or check whether the consumables are sufficient according to the instrument instruct...

Embodiment 3

[0053] Embodiment 3 Performance evaluation of the kit prepared by the present invention

[0054] (1) Analytical sensitivity

[0055] Use the 0-value calibrator as a sensitivity control product to measure 20 wells, calculate the mean (M) and standard deviation (SD) of the luminescence value, and calculate the concentration value of M+2SD according to the dose-response curve, and the result is for the analytical sensitivity.

[0056]

[0057] (2) Linear detection:

[0058] Dilute 3 high-value samples close to 240AU / ml into 6 concentrations according to 1:2, 1:4, 1:8, 1:16, 1:40, and samples with low-value concentrations must be close to the lower limit of the linear range ( 6AU / ml). The samples of each concentration were detected twice, and the average value was calculated, and the average value of the results and the dilution ratio were fitted with a straight line by the least square method, and the linear correlation coefficient R was calculated, and R≧0.99 was required....

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Abstract

The invention discloses a kit for detecting a human parvovirus IgM antibody. The kit comprises a B19-IgM magnetic particle suspension, a magnetic particle B19-IgM enzyme conjugate, a magnetic particleB19-IgM sample diluent, a magnetic particle B19-IgM calibration product, a light-emitting substrate and a concentrated washing solution, wherein a solid phase carrier in the B19-IgM magnetic particlesuspension is magnetic particles with the carboxylation particle size of 0.05-1 micron, a coated antibody is a mouse anti human IgM mu-chain monoclonal antibody, and an enzyme-labeled antigen of themagnetic particle B19-IgM enzyme conjugate is a horse radish peroxidase-labeled gene recombinant antigen. The kit has the advantages of being capable of carrying out detection analysis on a sample byutilizing a full-automatic chemiluminescent analyzer, simple and convenient to operate, short in detection time, accurate in detection result, wide in detection linear range and the like.

Description

technical field [0001] The invention relates to an in vitro diagnostic immunoassay technology, in particular to a kit for detecting human parvovirus IgM antibody. Background technique [0002] Human parvovirus B19 (HPVB19) is a linear single-stranded DNA virus with a genome size of 5.6 kb, classified in the genus Rhodovirus of the subfamily Parvovirinae. Blood products and children aged 2-7 are the main sources of infection of HPV B19 virus, and respiratory tract, blood products and vertical transmission from mother to child are the common routes of transmission. HPV B19 infection occurs worldwide, with varying rates of infection according to age and geographical distribution. About 15% of preschool children, 50% of adults and 85% of the elderly are seropositive. HPV B19 generally causes self-limiting disease, and the symptoms of infection vary with the host's immune function and hematological constitution. The most typical symptom of infected children is erythema contagi...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N33/543G01N21/76
CPCG01N21/76G01N33/54326G01N33/56983G01N33/6854G01N2333/015
Inventor 王万利王春霞陶占领刘功成渠海郑业焕付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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