Treponema pallidum specific antibody chemical light emitting detection kit and preparation method thereof

A technology for chemiluminescence detection and treponema pallidum, which is applied in the direction of chemiluminescence/bioluminescence, analysis by making materials undergo chemical reactions, and measurement devices, can solve the problems of inability to culture Treponema pallidum in vitro, high false positive rate of cardiolipin antibodies, inspection The positive rate is not high, and the detection time is shortened, the sensitivity is improved, and the precision is improved.

Inactive Publication Date: 2015-01-28
ZHONGSHAN HOSPITAL XIAMEN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Treponema pallidum cannot be cultured in vitro, and the positive rate of direct etiological dark-field microscopy is not high. Serological tests include cardiolipin antibody and specific antibody detection. The false positive rate of cardiolipin antibody is high and the sensitivity is low.

Method used

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  • Treponema pallidum specific antibody chemical light emitting detection kit and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] see figure 1 , the Treponema pallidum-specific antibody chemiluminescence detection kit is provided with an outer packing box 1, an alkaline phosphatase-labeled recombinant antigen bottle 2, a luminescent substrate bottle 3, a Treponema pallidum-specific antibody negative control bottle 4, and a Treponema pallidum-specific antibody bottle. 5 bottles of positive control substance, 6 bottles of washing solution, and 7 bottles of recombinant antigen-coated microwell plate; 2 bottles of alkaline phosphatase-labeled recombinant antigen, 3 bottles of luminescent substrate, 4 bottles of negative control substance specific antibody against Treponema pallidum, Treponema pallidum The specific positive control bottle 5, the washing solution bottle 6 and the recombinant antigen-coated microwell plate 7 are arranged in the outer packaging box 1; the alkaline phosphatase-labeled recombinant antigen bottle 2 is equipped with alkaline phosphatase-labeled recombinant antigen and luminesc...

Embodiment 2

[0048] The following shows the use of the Treponema pallidum antibody high-throughput detection kit to detect Treponema pallidum-specific antibodies in clinical specimens of patients:

[0049]1. Specimen processing: Serum: 5mL of venous blood, placed in a 37°C water bath for 30 minutes, centrifuged at 3000g for 10 minutes, and the supernatant was used as a test sample for later use.

[0050] 2. Adding samples: add 100 μL of samples to the reaction plate, and make blank, negative and positive control wells at the same time. Incubate at 37°C for 1h.

[0051] 3. Washing: After reacting at 37°C for 30 minutes, the measured syphilis-specific antibody binds to the syphilis-specific recombinant antigen coated on the microwell plate, and the unbound free components are separated by washing;

[0052] 4. Add alkaline phosphatase to label the recombinant antigen, then add luminescent substrate working solution, alkaline phosphatase catalyzes dephosphorylation of the substrate, and emits...

Embodiment 3

[0054] The performance test of the Treponema pallidum antibody high-throughput detection kit is given below.

[0055] (1) Conformity rate of positive samples

[0056] Use 50 positive reference sera of syphilis-specific antibodies to test, and calculate the positive coincidence rate.

[0057] (2) Negative specimen coincidence rate

[0058] Use 50 samples of syphilis-specific antibody-negative reference sera to test, and calculate the negative coincidence rate.

[0059] (3) Intra-batch variance

[0060] The same batch of kits is tested with characteristic serum, requiring CV ≤ 10%.

[0061] (4) Difference between batches

[0062] Different batches of kits are tested with characteristic serum, requiring CV ≤ 12%.

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Abstract

The invention discloses a treponema pallidum specific antibody chemical light emitting detection kit and a preparation method thereof, and relates to treponema pallidum. The kit comprises an outer package box, an alkaline phosphatase marked recombinant antigen bottle, a light emitting substrate bottle, a treponema pallidum specific antibody negative control bottle, a treponema pallidum specific antibody positive control bottle, a washing liquid bottle and a recombinant antigen coated micropore plate. The preparation method comprises the following steps: firstly, preparing reponema pallidum specific recombinant antigen and the recombinant antigen coated micropore plate, marking alkaline phosphatase of the recombinant antigen, further preparing a light emitting substrate, a washing liquid and a control group, and finally assembling the treponema pallidum specific antibody chemical light emitting detection kit. The treponema pallidum specific antibody chemical light emitting detection kit can be used for detecting syphilis specificity specific antibodies in clinical specimens, specific solid phase is adopted, immunity and light emitting reaction can be rapidly completed within a short time, the light emitting signals are greatly intensified, the sensitivity is improved, the detection time can be shortened, and the precision is improved.

Description

technical field [0001] The invention relates to Treponema pallidum, in particular to a Treponema pallidum-specific antibody chemiluminescence detection kit for detecting Treponema pallidum specific antibodies using a microwell reaction plate chemiluminescence method and a preparation method thereof. Background technique [0002] Syphilis is a sexually transmitted disease caused by Treponema pallidum. The incidence of syphilis has been increasing year by year in China in recent years. The prevention and control of syphilis has been listed as one of the main tasks of public health services in my country. [0003] Treponema pallidum cannot be cultured in vitro, and the positive rate of direct etiological dark-field microscopy is not high. Serological tests include cardiolipin antibody and specific antibody detection. The false positive rate of cardiolipin antibody is high and the sensitivity is low. The syphilis-specific antibody detection methods include treponemal pallidum ge...

Claims

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Application Information

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IPC IPC(8): G01N33/571G01N33/535G01N21/76
CPCG01N33/571G01N21/76G01N33/535
Inventor 刘莉莉杨天赐童曼莉张惠林林丽蓉张长弓
Owner ZHONGSHAN HOSPITAL XIAMEN UNIV
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