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A method for detecting combined immunization effect of pig spleen transfer factor and pseudorabies live vaccine

A technology for porcine spleen transfer factor and pseudorabies, which is applied in biochemical equipment and methods, measuring devices, biological testing and other directions, can solve the problems of high testing cost, long cycle, and large individual differences of organisms, and achieves high testing cost and results. Stable, low-cost test results

Active Publication Date: 2020-04-14
派生特(福州)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 current detection of the combined effect of transfer factor and live pseudorabies vaccine is mainly through the detection of antibodies and cytokines. The detection cost is high, the cycle is long, and the individual organisms vary greatly, and the detection results are unstable.

Method used

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  • A method for detecting combined immunization effect of pig spleen transfer factor and pseudorabies live vaccine
  • A method for detecting combined immunization effect of pig spleen transfer factor and pseudorabies live vaccine
  • A method for detecting combined immunization effect of pig spleen transfer factor and pseudorabies live vaccine

Examples

Experimental program
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Effect test

Embodiment 1

[0048] A method for detecting the combined immune effect of pig spleen transfer factor and live pseudorabies vaccine, comprising the following steps:

[0049] 1) Take live pseudorabies vaccine (Bartha-K61 strain) stock solution and dilute it with complete DMEM medium containing 2% newborn bovine serum and 1% penicillin-streptomycin double antibody to 200 TCID 50 virus liquid;

[0050] Take pig spleen transfer factor and dilute it with complete DMEM medium containing 2% newborn bovine serum and 1% penicillin-streptomycin double antibody to dilute the polypeptide content to 1 mg / mL;

[0051] 2) Culture pk-15 cells to monolayer in complete DMEM medium containing 10% newborn bovine serum and 1.0% penicillin-streptomycin double antibody;

[0052] 3) Wash the monolayer cells in step 2) twice with phosphate buffered saline without calcium and magnesium ions, then add 1.0mL trypsin digestion solution, shake the culture dish gently to make the digestion solution cover all the cell sur...

Embodiment 2

[0060] A method for detecting the combined immune effect of pig spleen transfer factor and live pseudorabies vaccine, comprising the following steps:

[0061] 1) Take live pseudorabies vaccine (Bartha-K61 strain) stock solution and dilute it with complete DMEM medium containing 1.8% newborn bovine serum and 0.8% penicillin-streptomycin double antibody to 200 TCID 50 virus liquid;

[0062] Take pig spleen transfer factor and dilute it with complete DMEM medium containing 1.8% newborn bovine serum and 0.8% penicillin-streptomycin double antibody to dilute the polypeptide content to 1 mg / mL;

[0063] 2) Culture pk-15 cells to monolayer in complete DMEM medium containing 8% newborn bovine serum and 0.8% penicillin-streptomycin double antibody;

[0064] 3) Wash the monolayer cells in step 2) twice with phosphate buffered saline without calcium and magnesium ions, then add 1.0mL trypsin digestion solution, shake the culture dish gently to make the digestion solution cover all the c...

Embodiment 3

[0072] A method for detecting the combined immune effect of pig spleen transfer factor and live pseudorabies vaccine, comprising the following steps:

[0073] 1) Take live pseudorabies vaccine (Bartha-K61 strain) stock solution and dilute it with complete DMEM medium containing 2.2% newborn bovine serum and 1.2% penicillin-streptomycin double antibody to 200 TCID 50 virus liquid;

[0074] Take pig spleen transfer factor injection and dilute it with complete DMEM medium containing 2.2% newborn bovine serum and 1.2% penicillin-streptomycin double antibody to dilute the polypeptide content to 1 mg / mL;

[0075] 2) Culture pk-15 cells to monolayer in complete DMEM medium containing 12% newborn bovine serum and 1.2% penicillin-streptomycin double antibody;

[0076] 3) Wash the monolayer cells in step 2) twice with phosphate buffered saline without calcium and magnesium ions, then add 1.0mL trypsin digestion solution, shake the culture dish gently to make the digestion solution cove...

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Abstract

The invention provides a method for detecting a combined immunization effect of a pig spleen transfer factor and a pseudorabies attenuated vaccine. The method comprises the following steps: (1) carrying out cell culture; (2) carrying out cell inoculation, and controlling cell density; (3) loading the pseudorabies attenuated vaccine and the pig spleen transfer factor into a cell culture plate, andculturing and hatching cells; (4) carrying out reaction on the cells by using an MTT solution, continuing to hatch, then carrying out dissolution treatment by using DMSO, finally putting the cell culture plate in a microplate reader for measuring an OD value, and calculating cell activity. The method provided by the invention has the advantages of low detection background, high sensitivity, strongspecificity, high accuracy, good repeatability and the like, and is particularly suitable for detection of bioactivity and establishment of quality standard during combined immunization of the pig spleen transfer factor and the pseudorabies attenuated vaccine.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for detecting combined immune effect of pig spleen transfer factor and pseudorabies live vaccine. Background technique [0002] Transfer Factor (TF) can regulate the immune function of animals, cooperate with vaccine immunization, shorten the period of vaccine immune response, and improve the level of animal immunity. It has a very broad application prospect. Transfer factor is a dialyzable small molecular substance present in human and animal leukocytes, which can transfer the cellular immune function of the donor to the recipient, thereby non-specifically enhancing the immune function of the recipient. The components of transfer factor mainly include oligonucleotides and polypeptides, which can induce the activation of immune cells in the body, enhance the immunity of the body, and transfer various antigens including viruses, bacteria, parasites, fungi,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02C12N5/071G01N33/50
CPCG01N33/5044G01N2800/52
Inventor 谢兆文谭礼宁吕小婷叶盛聪邱灵珊陈盛勇刘楚楚徐磊刘友霖
Owner 派生特(福州)生物科技有限公司