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A kind of detection method of interferon biological activity

A technology of biological activity and detection method, applied in biological testing, biological material analysis, measuring devices, etc., can solve the problems of inability to detect activity, large subjective error, poor sensitivity, etc.

Active Publication Date: 2019-03-01
GENSUN INST OF BIOMEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims to solve the problems of difficult observation, large subjective error, poor sensitivity and inability to detect activity due to species specificity in the existing interferon titer detection method when detecting porcine IFN-γ.

Method used

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  • A kind of detection method of interferon biological activity
  • A kind of detection method of interferon biological activity
  • A kind of detection method of interferon biological activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 The cultivation and titer titration of VSV

[0027] This example is only for illustration, not for limiting the present invention, and all conventional VSV culture and virulence determination methods can be used in the present invention.

[0028] (1), hatching of chicken embryo:

[0029] 1. Temperature: The optimum incubation temperature is 37.8°C.

[0030] 2. Humidity: generally 40% to 70%.

[0031] 3. The eggs should be placed with the big head up and the small head down. They cannot be placed flat, let alone hatched with the small head up. Approve the temperature and humidity of the incubator, and record the date and time of laying eggs. During the incubation process, attention should be paid to temperature and humidity, as well as ventilation and egg turning (generally 5 times a day; the angle of egg turning should be 45 degrees from the horizontal position forward and backward; when turning eggs, the movements should be light, steady, slow).

[00...

Embodiment 2

[0065] Example 2 Porcine interferon sample biological activity assay in different cell line detection systems:

[0066] (1) Method

[0067] 1. Pre-cultured IBRS-2 / WISH / PK-15 monolayer cells were digested with 0.05% Trypsin and then mixed with 10% FBS MEM / DMEM (MEM medium for IBRS-2 / WISH, DMEM medium for PK-15) ) to a suitable cell density (approximately 2-3×10 5 cells / mL) cell suspension, spread 100 μL / well of 96-well cell plate, 37 °C 5% CO 2 Cultivate for about 4 hours until it adheres to the wall.

[0068] 2. After the cells adhered to the wall, the porcine interferon α / γ sample was first diluted 4 times to make 12 gradients, each gradient was repeated for 3 wells, and diluted with 7% FBS MEM / DMEM medium. Add the diluted porcine interferon sample once to the adherent 96-well cell plate for medium replacement, 100 μL / well, add 100 μL dilution solution to the cell control and virus control wells, and store at 37°C in 5% CO 2 Incubate overnight for 18-24 hours until a unif...

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Abstract

The invention relates to a detection method of interferon biological activity. According to the detection method, pig kidney cells (IBRS-2) are taken as measuring cells, vesicular stomatitis virus (VSV) is taken as the challenge virus to detect interferon activity. The detection method can be applied to pig IFN-gamma titer measuring to solve problems in the prior art in pig IFN-gamma titer measuring that observation is difficult to realize, subjective error is large, sensitivity is poor, and activity detection is impossible to realize because of species specificity. The detection method can also be used for measuring of titer of other pig interferons such as IFN-alpha, and also can be used for measuring non-II-type human interferons such as human IFN-alpha.

Description

technical field [0001] The invention belongs to the field of immunology and microbiology, and relates to a method for detecting the biological activity of biological products, in particular to a method for detecting the biological activity of interferon. Background technique [0002] my country has a long history of raising pigs and is a big pig raising country in the world, with an annual stock of about 1 billion pigs. With the development of scale and intensification of the pig industry, pig diseases, especially viral diseases of pigs, have brought serious threats to the pig industry. In recent years, porcine circovirus (PCV2) disease and porcine reproductive and respiratory syndrome (PRRS) ) and other viral diseases, especially the immunosuppression caused by these virus infections, lead to the mixed infection of multiple pathogens in pig herds, and the death rate of pigs reaches 8%-10% or higher, thus causing huge economic losses every year. At present, although my coun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/6866G01N2333/57
Inventor 马永何青
Owner GENSUN INST OF BIOMEDICINE
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