Preparation method of sensitized chicken erythrocyte as well as IBV (Infectious Bronchitis Virus) detection kit
A technology for sensitizing chicken erythrocytes and chicken erythrocytes, which is applied in the field of preparation methods of detection materials and detection kits, can solve the problems of inaccurate test results, poor IB sensitivity, and increased detection costs, and achieve shortened detection time and easy Operation, low cost effect
Active Publication Date: 2014-01-29
GUANGZHOU SOUTH CHINA BIOLOGICAL MEDICINE
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The above serological detection methods may lead to inaccurate test results due to individual differences in the test subjects. Compared with IB, the agar diffusion test is less sensitive, and some serotypes are difficult to appear precipitation lines. This method is rarely used clinically for the detection of IB antibodies. Detection, HI and ELISA have high sensitivity, and the production cost will also increase accordingly. HI is more commonly used in clinical antibody detection. At present, there is no domestic manufacturer that provides better IB agglutination antigen. It needs to be purchased from Gezonheidsdienst Voor Dieren B.V. in the Netherlands. This greatly increases the cost of testing
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preparation example Construction
[0027] Preparation of fresh chicken red blood cells:
[0028] Take fresh blood from SPF chickens, centrifuge to remove serum, wash erythrocytes with pH 7.0 PBS solution for 3 to 4 times, centrifuge to precipitate erythrocytes, and store them at 4°C for later use.
[0029] Of course, other known methods can also be used to obtain chicken erythrocytes. Chicken red blood cells used in the following experiments
[0030] Solidification of chicken red blood cells:
[0031] Chicken erythrocytes were diluted to 8.48×10 with pH7.0 PBS 8 ~8.72×10 8 1 / ml, divided into 10 parts, adding glutaraldehyde respectively, adjusting its mass concentration to 0.075‰, 0.15‰, 0.3‰, 0.6‰, 1.25‰, 2.5‰, 5‰, 10‰, 15‰, 25‰, Take out by shaking at 25°C for 30 minutes, then wash the red blood cells with pH 7.0 PBS solution for 3 to 4 times, centrifuge to obtain solidified red blood cells, and store them at 4°C for later use;
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The invention discloses a preparation method of sensitized chicken erythrocyte as well as an IBV (Infectious Bronchitis Virus) detection kit. The preparation method of the sensitized chicken erythrocyte comprises the following steps of cell curing, wherein fresh chicken erythrocyte is washed by using buffer liquor, diluted by adding aldehyde liquor, oscillated for reaction and washed to obtain cured chicken erythrocyte; cell tanning, wherein the cured chicken erythrocyte is diluted, and added with tannic acid to obtain tanned chick erythrocyte; and cell sensitization, wherein the tanned chicken erythrocyte is diluted, added with IBV liquor for mixing and acting to obtain the sensitized chicken erythrocyte. The sensitized chicken erythrocyte disclosed by the invention has very high specificity, and a detection result has very good conformance. An IHA (Indirect Hemagglutination Assay) agent prepared and obtained by the sensitized chicken erythrocyte has low cost and is easy to operate, so that the result can be observed within 25 minutes and detection time is greatly shortened.
Description
technical field [0001] The invention relates to a preparation method of a detection material and a detection kit thereof, in particular to a preparation method of sensitized chicken red blood cells used for indirect hemagglutination detection of IBV antibodies and an IBV detection kit. Background technique [0002] Infectious bronchitis (IB) is an acute, highly contagious viral respiratory and genitourinary disease of chickens. It is characterized by coughing, sneezing, tracheal rales, and serous catarrhal inflammation of the respiratory mucosa. The prevalence of this disease is worldwide, and it can infect chickens of all breeds and ages, leading to growth retardation, death, weight gain and reduced feed remuneration of sick chickens. The quality of the laying hens has declined significantly, and the egg production of the recovered laying hens is difficult to return to the level before the disease. The disease often causes secondary infections such as Mycoplasma and E. Hu...
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IPC IPC(8): G01N33/531G01N33/569
CPCG01N33/531G01N33/56966G01N33/86G01N2469/00
Inventor 王鑫叶贺佳罗开健许丽娜李敏仇微红梁昭平
Owner GUANGZHOU SOUTH CHINA BIOLOGICAL MEDICINE
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