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Serum pre-dilution method for HI test

A pre-dilution, serum technology, applied in the field of HI experiments, can solve problems such as increasing the detection cost of HI experiments, and achieve the effects of improving serum utilization, reducing retest rate, and saving reagents

Inactive Publication Date: 2019-07-12
BEIJING POULTRY BREEDING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the 5 routine tests, due to the need to conduct multiple sets of experiments and provide data support for the final test results in the form of data comparison, there are many operations for adding samples. For dilution, about 400 sample addition actions are required, which delays the efficiency of the HI experiment and increases the detection cost of the HI experiment

Method used

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  • Serum pre-dilution method for HI test
  • Serum pre-dilution method for HI test

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A HI test serum pre-dilution method comprises the following steps.

[0029] S1: Dilute the serum to be tested by 8 times to form a sample to be tested.

[0030] The following steps take a 96-well reaction plate as an example. The specification of the reaction plate is not limited here, but it is required that the reaction plate has at least four rows with 12 wells in each row.

[0031] S2: Add 30 microliters of phosphate buffer solution to each well of the reaction plate, then select 1 row in order to add 30 microliters of phosphate buffer solution to each well as a negative control, and select 1 row to add 30 microliters of standard samples were used as positive controls, and 30 microliters of samples to be tested were added to each well of the remaining six horizontal rows.

[0032] S3: Doubling dilution: Use a pipette to repeatedly blow the first vertical hole 8 times, absorb 30 microliters of the test solution in the first vertical hole and add it to the second ver...

Embodiment 2

[0038] The difference between this example and Example 1 is that in step S2, after adding 30 microliters of phosphate buffered saline to each well, select one horizontal row and add 30 microliters of phosphate buffered saline to each well in order to make For negative control, add 30 microliters of 8-fold diluted standard sample to each well of 1 horizontal row, select 1 horizontal row and add 30 microliters of standard sample to each well of positive control, and add 30 microliters of sample to be tested to each well of the remaining five horizontal rows. Since the 8-fold diluted standard sample was added, the 8-fold diluted positive control was added, thereby reducing data errors due to dilution.

Embodiment 3

[0040] The difference between this example and Example 1 is that in step S2, after adding 30 microliters of phosphate buffered saline to each well, select one horizontal row and add 30 microliters of phosphate buffered saline to each well in order to make Negative control, choose 1 horizontal row and add 30 microliters of 8-fold diluted standard samples to each well, choose 1 horizontal row and add 30 microliters of standard samples to each well as positive control, choose 4 horizontal rows and add 30 microliters of samples to be tested in each well , Add 30 microliters of the serum to be tested to each well of the remaining 1 horizontal row, that is, the sample to be tested before 8-fold dilution. Since the 8-fold diluted standard sample was added, the 8-fold diluted positive control was added, thereby reducing data errors due to dilution. Due to the addition of the serum to be tested, a set of normal HI test data is provided. It can not only provide the experimental data und...

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Abstract

A serum pre-dilution method for an HI test comprises the following steps: diluting serum to be detected by 8 times to form a sample to be detected; adding an equal amount of phosphate buffer solutioninto each hole of a reaction plate, and then adding a positive-negative control and the sample to be detected which are equal to the phosphate buffer solution in amount; carrying out double-ratio dilution; adding an antigen solution into each hole; adding 1% of erythrocyte suspension into each hole; expanding the interpretation range to 4-15 times, and correspondingly reading the final data of thereaction plate. The serum to be detected is diluted by 8 times in advance; according to the serum pre-dilution method for an HI test, after an HI experiment pre-dilution is carried out, the concentration of a sample in each hole of the reaction plate is reduced by 8 times, and the original interpretation range is expanded by 8 times, so that a re-detection step is avoided, operation steps in theHI experiment serum pre-dilution process are reduced, and the detection cost of an HI test sample is reduced.

Description

technical field [0001] The invention relates to the technical field of HI experiments, in particular to a serum pre-dilution method for HI experiments. Background technique [0002] The HI test is the hemagglutination inhibition test, which is a common method for measuring serum antibodies to hemagglutination antigens such as avian influenza, Newcastle disease, and egg drop syndrome. All poultry disease laboratories and research units must use this method. [0003] Most of the current HI routine monitoring methods include the following steps: sample addition and doubling dilution, addition of antigen and antigen-body reaction, addition of red blood cell suspension reaction, and reading the results. For example, for a 96-well hemagglutination plate (reaction plate), it contains 8 rows with 12 holes in each row. During the experiment, add corresponding PBS, negative and positive controls, samples to be tested, etc. to each row. The device realizes the doubling dilution, so as...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/86G01N1/38
CPCG01N1/38G01N33/53G01N33/5306G01N33/86
Inventor 梁宁王海舰郑云平
Owner BEIJING POULTRY BREEDING
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