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A pig anti-human T cell enzyme-linked immunoassay kit and detection method thereof

An immunodetection and cellular enzyme technology, applied in the field of kits, can solve the problems of complicated operation steps, long time-consuming, large error in experimental results, etc., and achieve the effects of fast detection process, simple operation process and strong accuracy.

Inactive Publication Date: 2011-12-28
湘潭陆峰生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the operation steps of the rosette test are cumbersome, and the experimental conditions require a constant temperature of 22°C, which is difficult and has large errors in the experimental results. For example, the manual counting of rosettes and lymphocytes is difficult, affected by subjective factors, and the accuracy is poor. Secondly, the experimental operation generally takes 2 hours. about days, time-consuming

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  • A pig anti-human T cell enzyme-linked immunoassay kit and detection method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1: Preparation of enzyme-labeled antigen: take isolated and purified human T cells and dilute to 5×10 6 After each milliliter, put it in a -20°C refrigerator for 20 minutes, take it out and put it in a 20°C constant temperature water bath for 10 minutes, repeat 3 times, pour it into a centrifuge tube, centrifuge at 10,000rpm for 5 minutes, and take the supernatant as the enzyme-labeled antigen.

Embodiment 2

[0022] Example 2: Preparation of the pig anti-human T cell ELISA kit: The pig anti-human T cell ELISA kit includes a coated ELISA plate, enzyme-labeled antigen, anti-pig ELISA antibody, coating solution, Washing solution, diluting solution, blocking solution, phosphate-citrate buffer solution, substrate solution, terminator, the anti-pig enzyme-labeled antibody includes standard positive serum and standard negative serum.

[0023] NA in every 1000mL coating solution in the above pig anti-human T cell ELISA kit 2 CO 3 Contains 1.59g, NAHCO 3 Contains 2.93g, distilled water 1000mL, pH of the coating solution is 9.6; NACL content per 1000mL of washing solution is 8.2 g, KH 2 PO 4 0.2 g, NA 2 HPO 4 12H 2 O is 2.9 g, KCL is 0.2 g, Tween-20 is 0.5 mL, distilled water is 999.5 mL; BVDV antibody-negative calf serum or rabbit serum is 100 mL per 1000 mL of dilution, bovine serum albumin is 10 mL, and washing solution is 890 mL; per 1000 mL Bovine serum albumin 10mL in blocking s...

Embodiment 3

[0024] Embodiment 3: In this embodiment, the method for detecting serum with the pig anti-human T cell ELISA kit is carried out according to the following steps:

[0025] a. Dilute the enzyme-labeled antigen 1000 times in the coating solution, coat the enzyme-labeled plate, add 100 μL of the enzyme-labeled antigen diluent to each well, leave it at 4 overnight, and act for 24 hours;

[0026] b. Add 100 μL of blocking solution to each well and place it at 37°C for 1 hour. The sealed microplate can be stored at -20°C for 12 months at low temperature;

[0027] c. Wash the microtiter plate with washing solution, shake off the washing solution, and then dry;

[0028] d. Dilute the serum to be tested with diluent, standard positive serum and standard negative serum 50 times; add different serum diluents to the wells of the microtiter plate and mark them, and incubate at 37°C for 45 minutes:

[0029] e. Washing as in step c;

[0030] f. The diluent dilutes the enzyme-labeled an...

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Abstract

The invention discloses a pig anti-human T cell ELISA detection kit and a detection method thereof. The kit includes coated enzyme-labeled plate, enzyme-labeled antigen, anti-pig enzyme-labeled antibody, coating liquid, washing liquid, diluting liquid, blocking liquid, phosphate-citrate buffer, substrate solution, and terminator. The detection method comprises the following steps: coating the enzyme-labeled antigen, sealing the enzyme-linked plate, washing, fixing the standard serum and the sample to be tested in the enzyme-linked plate, detecting the light absorption value, and finally analyzing the sample index. The invention has the advantages of simple operation process, rapid detection process and high accuracy.

Description

technical field [0001] The invention relates to a kit, in particular to a swine anti-human T cell enzyme-linked immunoassay kit and a detection method thereof. Background technique [0002] At present, the method for detecting pig anti-human T cell immunoglobulin indicators is mainly: "Pharmacopoeia of the People's Republic of China" Appendix XQ Anti-human T cell immunoglobulin titer determination method, referred to as the inhibitory rosette test, and the control inhibitory titer is greater than or equal to 1 than 1000. However, the operation steps of the rosette test are cumbersome, and the experimental conditions require a constant temperature of 22°C, which is difficult and has large errors in the experimental results. For example, the manual counting of rosettes and lymphocytes is difficult, affected by subjective factors, and the accuracy is poor. Secondly, the experimental operation generally takes 2 hours. About a day, it takes a long time. Contents of the inventi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 张勤
Owner 湘潭陆峰生物科技有限公司