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Treponema pallidum antibody detection kit

A technology for detecting Treponema pallidum and antibodies, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low retest rate and achieve the effect of automatic detection and improvement of false positive problems

Inactive Publication Date: 2020-06-19
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there is an urgent need to provide a low retest rate Treponema pallidum antibody detection reagent and detection method

Method used

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  • Treponema pallidum antibody detection kit

Examples

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Effect test

Embodiment 1

[0038] The preparation of embodiment 1 Treponema pallidum (TP) antibody detection kit

[0039] 1. Preparation of TP antigen-coupled magnetic particles

[0040] 1.1 Magnetic particle washing

[0041] Take 200 μL of magnetic particles containing carboxyl groups on the surface and put them in a glass bottle, use a magnet to adsorb the magnetic particles to the bottom of the glass bottle, remove the supernatant; add 2 mL of PBS (pH 7.4), and repeat the above operation 3 times.

[0042] 1.2 Activation of Magnetic Particles

[0043]Dissolve EDC and NHS in 0.1MMES (pH 5.0) buffer solution at a concentration of 10-70 mg / mL, and then add 1 mL each to the magnetic particles; shake gently at room temperature for 30-60 minutes; use a magnet to shake the magnetic particles Adsorbed at the bottom, remove the supernatant; then add 2mL 0.1MMES (pH5.0) buffer to resuspend the magnetic particles; repeat the above operation 2 times.

[0044] 1.3 TP antigen coating

[0045] Add 0.1MMES (pH5.0...

Embodiment 2

[0053] The detection method of embodiment 2 Treponema pallidum (TP) antibody detection kit

[0054] The kit prepared in Example 1 is used in conjunction with the automatic chemiluminescence instrument AutoLumo A2000 or AutoLumo A2000 Plus produced by Zhengzhou Antu Bioengineering Co., Ltd. to detect Treponema pallidum (TP) antibodies:

[0055] Add 2 negative control wells and 3 positive control wells (for determining the Cutoff value) in sequence in the reaction container (hereinafter referred to as "well"), 50 μL / well; add 50 μL sample to each well of the remaining wells;

[0056] Add 20 μL of magnetic particle suspension and 50 μL of enzyme conjugate to each well; after mixing, incubate at 37°C for 30 minutes; then wash 5 times with cleaning solution;

[0057] Add 50 μL each of Substrate A and Substrate B to each well; measure the luminescence intensity 1 to 5 minutes after mixing;

[0058] Calculation of results: Cutoff value = average luminescence value of positive contro...

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Abstract

The invention relates to the technical field of in vitro diagnosis, in particular to a treponema pallidum antibody detection kit. The kit comprises treponema pallidum antigen coupling magnetic particles, a treponema pallidum antigen enzyme conjugate and a chemiluminiscence substrate, the treponema pallidum antigen enzyme conjugate is a buffer solution containing a treponema pallidum antigen markedby a biological enzyme, urokinase and calf serum. According to the invention, magnetic particles are used as a solid-phase carrier of the antibody, and a chemiluminescence immunoassay technology is utilized, so that single-person, random, rapid and automatic detection can be realized on a full-automatic chemiluminescence analyzer. Tests prove that adding 0.1-30U / mL of urokinase into the enzyme conjugate has no influence on indexes such as sensitivity, specificity, stability, precision and the like, but the clinical retest rate is greatly improved.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a Treponema pallidum antibody detection kit. Background technique [0002] Syphilis is a chronically transmitted disease caused by Treponema pallidum (TP). In the route of transmission of syphilis, sexual contact infection accounts for 95%, and it is mainly transmitted from the damaged part through sexual intercourse. TP exists in large quantities on the surface of skin and mucous membrane damage, and is also found in saliva, milk, semen and urine. Untreated patients are most contagious within one year of infection. With the prolongation of the disease period, the infectivity gradually weakens. Those with a disease period of more than 4 years are not contagious through sexual contact. [0003] Treponema pallidum (Treponema pallidum, TP) belongs to Spirochaetales (Spirochaetales) Spirochaetaceae (Spirochaetaceae) Treponema (Treponema), is a kind of treponema, in the fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/571G01N33/535G01N33/543G01N21/76
CPCG01N21/76G01N33/535G01N33/54326G01N33/571
Inventor 江雪王新明孙冯博娄本光
Owner AUTOBIO DIAGNOSTICS CO LTD
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