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Kit for detecting human HTLV virus (1+2) type antibody

A kit and virus technology, applied in the field of kits for detecting human HTLV virus (1+2) antibodies using magnetic particle technology, can solve the problems that PCR cannot be detected with stock serum, complicated PCR operation, unsuitable screening reagents, etc., to achieve Reliable detection results, high detection sensitivity, and low degree of automation

Inactive Publication Date: 2019-01-11
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PCR has high sensitivity and specificity, and is often used in the evaluation of other HTLV-Ⅰ / Ⅱ detection methods, but the operation of PCR is complicated, the cost is high, and it is easy to contaminate and cause false positives, so it is not suitable for routine use
Therefore, it is mostly used as a confirmatory test for HTLV infection at present, and is not suitable as a screening reagent.
Also, since viremia is rare in HTLV-infected individuals, PCR cannot be used for testing banked sera
[0007] 2. Enzyme-linked immunosorbent assay (ELISA) and gelatin particle agglutination (PA): This is a preliminary screening test for the detection of HTLV-Ⅰ / Ⅱ antibodies. The most commonly used antigens are gp46, p2le, p24 and p40tax. This method is sensitive High, but low specificity, suitable for large-scale primary screening identification
In order to solve the problem of lack of large-scale detection reagents in my country's HTLV research, it is very necessary to invent a kit for detecting human HTLV virus type I and type II antibodies using magnetic particle technology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Preparation of a kit for detecting antibodies to human HTLV virus (1+2)

[0025] 1. Preparation of magnetic particle coating suspension

[0026] The first step is to mix and suspend the magnetic beads, take 1 volume of carboxy-modified magnetic beads and wash them with 70 volumes of buffer solution (0.02mol / L PBS buffer), then place them on a magnet to separate the magnetic beads;

[0027] In the second step, the separated magnetic beads are activated by adding dissolved 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride solution;

[0028] In the third step, the activated magnetic beads in the second step are incubated with glacial acetic acid solution at pH 5.5 and 1 volume of HTLV (1+2) fusion antigen at room temperature;

[0029] The fourth step is to separate the magnetic beads obtained in the third step, and block the activity of the carboxyl group on the magnetic beads with ethanolamine;

[0030] The fifth step is to separate the magnetic bead...

Embodiment 2

[0037] The detection method of the kit prepared in embodiment 2 embodiment 1

[0038] 1. Sample: HTLV national reference product;

[0039] 2. Establish a reaction system: 20 μl of magnetic particle coating suspension, 100 μl of enzyme conjugate working solution, 30 μl of sample diluent, 50 μl of luminescent substrate A solution, and 50 μl of luminescent substrate A solution.

[0040] 3. Detection method:

[0041] Take 100 μl of sample + 20 μl of magnetic particle coating suspension + 30 μl of sample diluent, react at 37°C for 15 minutes, wash, add 100 μl of enzyme conjugate, react at 37°C for 15 minutes, wash, add 50 μl each of luminescence substrate solution A and B, and react at 37°C 1-5min to detect the luminescence value.

Embodiment 3

[0042] The performance test of the kit prepared in embodiment 3 embodiment 1

[0043] The performance test was carried out with the national reference product of HTLV antibody.

[0044] National reference requirements: Negative reference product compliance rate: no positive reaction, negative compliance rate (- / -) should be 16 / 16; HTLV-1 positive reference product compliance rate: all positive reactions, HTLV-1 type The positive coincidence rate (+ / +) should be 8 / 8; the coincidence rate of HTLV-2 positive reference products: all should be positive reactions, and the positive coincidence rate (+ / +) of HTLV-2 should be 2 / 2; Out-of-limit reference product: no less than 3 positive reactions (≥3 / 6) and negative reaction in matrix serum (S1); precision reference product: 10 parallel tests, and the coefficient of variation (CV value) should be ≤15%. Can meet the national disk requirements.

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Abstract

The invention discloses a kit for detecting a human HTLV virus (1+2) type antibody, comprising a magnetic particle coating suspension, an enzyme conjugate working solution, and a sample diluent; the coating antigen is a HTLV (1+2) fusion antigen; and the enzyme conjugate is a horseradish peroxidase-labeled HTLV (1+2) fusion antigen. According to the kit for detecting a human HTLV virus (1+2) typeantibody, the kit of the invention adopts magnetic particles as a solid-phase carrier and has a large specific surface area, which increases the area where the immune reaction occurs and improves thesensitivity of the reaction; and the spherical and uniform surface of the magnetic particles reduce chemical adhesion and non-specific binding, enhance a specific enzymatic chemiluminescence system toachieve effective amplification of a reaction signal. The kit of the invention is good in specificity, so that the antigen and the antibody achieve high detection sensitivity, and a fully automatic measurement is achieved. The kit of the invention can be used with a fully automatic chemiluminometer, and the influence of human operation on test results is reduced, so that the detection result is more reliable, accurate, fast and repeatable.

Description

technical field [0001] The invention relates to a magnetic particle detection technology, in particular to a kit for detecting human HTLV virus (1+2) type antibody by using the magnetic particle technology. Background technique [0002] Human T-lymphotropic virus (HTLV) is the earliest discovered human retrovirus, mainly including four subtypes, among which HTLV-1 and HTLV-2 are 70% homologous at the gene level. Transfusion-transmitted HTLV-1 / 2 mainly causes HTLV-1 / 2-associated myelopathy, tropical spastic paraparesis (TSP) and myelopathy (HAM). Infection is generally asymptomatic, and there is transmission from close contacts; blood transfusion infection HTLV-1 has a short incubation period, and the onset of HTLV-1 can occur 1 to 4 months after transfusion of infected blood, while natural infection usually occurs after several years or even several years. Ten years of illness. [0003] In order to ensure blood safety and prevent the transmission of HTLV-1 / 2 through blood ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543G01N33/569G01N33/68G01N21/76
Inventor 吴文娟王新明孙冯博刘功成付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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