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Hydroformylated fresh goose red blood cells and application of hydroformylated fresh goose red blood cells to hemagglutination (HA) test for acellular pertussis

A technology of erythrocytes and goose hydroformylation, applied in the field of acellular pertussis detection technology, can solve the problems of short storage period, brittle and fragile quality, and achieve accurate test results

Inactive Publication Date: 2012-10-03
WUHAN INST OF BIOLOGICAL PROD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the HA test, the 1% goose fresh red blood cells that need to be used have the disadvantages of brittle quality and short shelf life

Method used

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  • Hydroformylated fresh goose red blood cells and application of hydroformylated fresh goose red blood cells to hemagglutination (HA) test for acellular pertussis
  • Hydroformylated fresh goose red blood cells and application of hydroformylated fresh goose red blood cells to hemagglutination (HA) test for acellular pertussis
  • Hydroformylated fresh goose red blood cells and application of hydroformylated fresh goose red blood cells to hemagglutination (HA) test for acellular pertussis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment １

[0028] Preparation of Aldehylated Goose Fresh Red Blood Cells by the First Method of Aldehydating Goose Fresh Red Blood Cells

[0029] The preparation method of the fresh erythrocytes of goose of formaldehyde comprises the following steps:

[0030] Step 1: Use a syringe to draw about 2-6 mL of goose venous blood, put it into a centrifuge tube filled with 10 mL of normal saline and mix well, centrifuge at 1500-2000 r / min 4 °C for 8 minutes, discard the supernatant, and obtain a precipitate Add 10 mL of normal saline to the precipitate, mix well, and centrifuge at 1500-2000 r / min for 8 minutes, remove the supernatant and the white blood cell film on the upper layer of the precipitated red blood cells with a pipette to obtain the precipitate, add 10 mL of normal saline to the precipitate, Mix well, then centrifuge at 1500-2000 r / min for 8 minutes, use a pipette to remove the supernatant and precipitate the white blood cell film on the upper layer of red blood cells, and the sedim...

Embodiment ２

[0035] Preparation of Aldehylated Goose Fresh Red Blood Cells by the Second Method of Aldehylated Goose Fresh Red Blood Cells

[0036] Step 1: Same as Example 1

[0037] Step 2: Slowly add 15 mL of 5% goose erythrocyte suspension to 15 mL of 2.5% glutaraldehyde saline solution, place in a refrigerator at 4°C and stir with a magnetic stirrer for 1 hour, take 4 mL, wash with normal saline five times, each time at 2000 r / min, centrifuge for 10 min, discard the supernatant, and make a 5% suspension. Store it at 4°C for later use, and obtain the hydroformylated goose erythrocytes that have been hydroformylated for 1 hour. Continue to stir the remaining solution for 2 h, 5 h, and 6 h respectively. According to the above method, take out part and wash it, make a 0.8% suspension, and store it at 4°C for later use. The hydroformylated goose erythrocytes were obtained after hydroformylation for 2h, 5h, and 6h respectively.

Embodiment ３

[0039] HA test for acellular pertussis

[0040]The HA test of acellular pertussis was carried out with the alcoholized goose fresh red blood cells prepared in Example 1, the alcoholized goose fresh red blood cells prepared in Example 2, and the 0.8% fresh goose red blood cell suspension belonging to the prior art.

[0041] Main Materials and Equipment

[0042] Bacillus pertussis (cultivated in a large tank in the department), healthy goose blood, 25% glutaraldehyde, high-speed refrigerated centrifuge (Eppendorf), electronic balance (Mettler), 96-well plate.

[0043] Specific process:

[0044] Preparation of fresh goose erythrocytes and determination of optimal hydroformylation time:

[0045] The first method of hydroformylation: Take 1 mL of fresh red blood cells from precipitated goose, slowly add 40 mL of 1.25-5% glutaraldehyde solution pre-cooled at 4°C, shake quickly, place in a refrigerator at 4°C and stir with a magnetic stirrer, and react for 40 minutes Take out 20mL...

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Abstract

The invention provides hydroformylated fresh goose red blood cells, a preparation method thereof and application of the hydroformylated fresh goose red blood cells to hemagglutination (HA) test for acellular pertussis. Specific conditions of the method for hydroformylating goose red blood cells are that: the goose red blood cells are hydroformylated for 40min by using 1.25 percent glutaraldehyde; the working concentration of hydroformylated goose red blood cells is 0.8 percent; and when the hydroformylated goose red blood cells are used for the HA test for acellular pertussis, a test result which is consistent with a test result obtained by using 0.8 percent fresh goose red blood cells is obtained, but after the fresh goose red blood cells are replaced by the hydroformylated goose red blood cells, the preservation period of the goose red blood cells is prolonged, experimental results are relatively stable, and the disadvantages that 1 percent fresh goose red blood cells which are required to be used in the prior art are crispy and fragile and the preservation period is short are overcome.

Description

technical field [0001] The invention relates to a method for detecting acellular pertussis. For acellular pertussis hemagglutination test HA (a certain virus or virus hemagglutinin), it can selectively agglutinate red blood cells of one or several animals. The phenomenon of agglutination of red blood cells is called hemagglutination test. The goose fresh red blood cells used in HA) are glutarylated to improve the problem of brittle goose red blood cells and short shelf life. Compared with goose fresh red blood cells, the shelf life is from 7 days to 10 days. The left and right are increased to 90 days, which reduces the frequency of collecting goose blood and reduces the work intensity. Background technique [0002] Whooping cough (pertussis, whooping cough) is an acute respiratory infectious disease in children. The typical symptoms are sudden paroxysmal spasmodic cough with an inspiratory end or accompanied by vomiting. It can last for several months. If there is no compli...

Claims

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Application Information

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IPC IPC(8): C12N5/078C12Q1/04
Inventor 赵伟胡业勤朱红伟薛红刚李新国项美娟
Owner WUHAN INST OF BIOLOGICAL PROD CO LTD
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