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Kit for detecting mastitis of dairy cow and using method thereof

A technology of detection reagents and kits, which is applied in the field of in vitro diagnosis of animal epidemics, can solve the problems of long detection time and unsuitability for single sample detection, and achieve the effect of low cost

Inactive Publication Date: 2019-12-27
BIOLOGY INST OF HEBEI ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The existing milk serum amyloid A detection product only has an ELISA detection kit, which takes a long time to detect and is not suitable for single sample detection

Method used

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  • Kit for detecting mastitis of dairy cow and using method thereof
  • Kit for detecting mastitis of dairy cow and using method thereof
  • Kit for detecting mastitis of dairy cow and using method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Preparation of Bovine Serum Amyloid A Fluorescence Immunochromatography Detection Kit

[0024] 1) Coating:

[0025] Paste the nitrocellulose membrane in the middle of the bottom plate, bovine serum amyloid A monoclonal antibody (purchased from Finland Medix

[0026] Biochemica company) use 0.01M PB buffer to dilute to 2mg / ml, use 0.01M PB buffer to dilute to 2mg / ml with rabbit anti-chicken IgY antibody (purchased from Nanjing Hanrui Boke Biotechnology Co., Ltd.) as T-line coating solution / ml, as the C-line coating solution. Coat the T-line and C-line coating solutions on the nitrocellulose membrane by using a three-dimensional gold-spraying scriber. After the film is scratched, it is dried in a blast drying oven at 45°C for 1 hour.

[0027] 2) Preparation of fluorescent microsphere-labeled bovine serum amyloid A monoclonal antibody:

[0028] a) Take 1 mg of fluorescent microspheres and dilute to 1 mL with 50 mM MES buffer pH 6.0;

[0029] b) Take about ...

Embodiment 2

[0050] Example 2: Drawing the calibration curve of the bovine serum amyloid A fluorescent immunochromatography detection kit

[0051] Use the 250mg / L sample valued by the bovine serum amyloid ELISA kit of Shanghai Lanji Biotechnology Co., Ltd. as a high-value reference product, and the sample is diluted with the sample diluent in the kit of the present invention to obtain 0mg / L, 0.4mg / L, 2mg / L, 10mg / L, 50mg / L, 100mg / L calibrator concentration points. Use the test paper card in the test kit of the present invention to carry out 10 repeated tests for each calibrator concentration point, and carry out four-parameter fitting with the T / C mean value and theoretical concentration value measured at each point of the calibrator, and draw the calibration curve( image 3 ), where the X-axis is the calibrator concentration point, and the Y-axis is the T / C mean.

Embodiment 3

[0052] Embodiment 3: The blank limit test of bovine serum amyloid A fluorescent immunochromatography detection kit

[0053] According to relevant literature reports, the sensitivity of the kit was evaluated by the test blank limit method.

[0054] Use the test paper card in the test kit of the present invention to test 20 times of 0 mg / L calibrator concentration point, calculate the T / C average value and standard deviation of this point, and substitute the numerical value of this (average value+2 times standard deviation) into the fixed value According to the standard curve equation, the calculated blank limit is 0.052mg / L. It shows that the test kit of the present invention tests samples with milk serum amyloid A content>0.052mg / L, and the detection result will be>0mg / L.

[0055] It has been reported in the literature that the reference value range of serum amyloid A content in milk samples is <0.55 mg / L, which means that the content of serum amyloid A in milk samples from h...

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Abstract

The invention relates to a fluorescence immunochromatography assay kit and a using method thereof, and belongs to the technical field of products for diagnosing animal epidemic diseases in vitro. Thekit comprises a plastic card shell, a detection reagent card and sample diluent, wherein the card shell comprises a bottom shell and an upper cover, a test strip clamping groove is formed in the bottom shell, and a scanning window and a sample adding hole are formed in the upper cover; the detection reagent card consists of a sample pad, a combination pad, a nitrocellulose membrane and absorbent paper which are sequentially adhered to a bottom plate; and the position of the scanning window is matched with that of the nitrocellulose membrane, and the position of the sample adding hole is matched with that of the sample pad. The kit provided by the invention can be used for judging whether a dairy cow suffers from mastitis or the severity of mastitis of the dairy cow by detecting the contentof serum amyloid A in milk, and has the characteristics of rapidness, accuracy and low cost.

Description

technical field [0001] The invention relates to a fluorescence immunochromatography assay kit and a use method thereof, in particular to a fluorescence immunochromatography assay kit for detecting cow mastitis and a use method thereof, and belongs to the technical field of in vitro diagnosis of animal epidemic diseases. Background technique [0002] Mastitis is a common disease in the process of raising dairy cows. Once a cow gets sick, it will seriously affect the quality of milk. If it is not possible to detect whether a cow has mastitis accurately and in time, it will cause huge economic losses to milk stations and dairy companies. . At present, mastitis is mainly screened by somatic cell count (SCC), but the sensitivity and accuracy of SCC are limited, and it is easily affected by other factors, so it cannot accurately reflect the inflammatory state. At present, there are few researches on the diagnosis of dairy cow mastitis in China. It is an urgent need to develop dai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/58G01N33/558G01N33/68
CPCG01N33/54306G01N33/54393G01N33/582G01N33/585G01N33/558G01N33/6893G01N2800/365G01N33/54388
Inventor 吴萌魏治静李春生陈英珠
Owner BIOLOGY INST OF HEBEI ACAD OF SCI
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