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Babesia caballi disease immunoblotting detection method and preparation of kit

A technology for babesia cerevisiae and immunoblotting, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems that there are no research reports on the detection method of babesia cerium ii immunoblotting, achieve short preparation cycle, shorten the cycle, The effect of using less reagent

Inactive Publication Date: 2011-05-18
中华人民共和国天津出入境检验检疫局
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There are very few domestic studies on the detection of animal diseases by Western blot, and there is no research report on the detection method of Babesia spp.

Method used

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  • Babesia caballi disease immunoblotting detection method and preparation of kit
  • Babesia caballi disease immunoblotting detection method and preparation of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Preparation of Western Blot Diagnostic Kit for Babesia cerevisiae

[0054] Preparation of reagents

[0055] 1. 1x VYM's buffer:

[0056] CaCl 2 .2H 2 O 0.016 g

[0057] KCl 0.400g

[0058] K H 2 PO 4 1.415g

[0059] MgSO 4 .7H 2 O 0.154 g

[0060] Na 2 HPO 4 0.077g

[0061] NaCl 7.077g

[0062] Glucose 20.500 g

[0063] wxya 2 O 1 L

[0064] After stirring and dissolving, add 0.0423 g of Adenine and 0.0708 g of Guanosine, adjust the pH to 7.0-7.2, vacuum filter (0.22 μm), and store at 4°C for later use.

[0065] 2. HL-1 tissue culture medium:

[0066] HL-1 medium 400 mL

[0067] Horse Serum 100 mL

[0068] Hepes 0.238 g

[0069] 2 mmol L-glutamine 500 μL

[0070] Antibiotic-antimycotic (100x) 1 mL

[0071] Stir magnetically for 5 min, adjust the pH to 7.2, vacuum filter (0.22 μm), and store at 4°C for later use

[0072] 3. PI buffer:

[0073] Tris 3.03g

[0074] Deionized water 500...

Embodiment 2

[0099] Example 2 : Western blot detection of babesia

[0100] Reagent preparation:

[0101] 1. 10x electrophoresis buffer:

[0102] Tris 121.1g

[0103] Glycine 570 g

[0104] Distilled water 4 L

[0105] 2. Transfer buffer:

[0106] 10x Running Buffer 100 mL

[0107] Methanol 200 mL

[0108] Distilled water 700 mL

[0109] 3、 blocking solution ： Phosphate buffered saline containing 0.2 % Tween-20, 10 % skimmed milk powder

[0110] 4. Lotion ： Phosphate buffered saline with 0.2 % Tween-20

[0111] Take a NuPAGE 4%-12% Bis-Tris gel plate, wash the gel well three times with 1 x SDS Running Buffer; place the gel plate face forward in the core frame of the Mini-Cell vertical electrophoresis instrument, and lock it. Add 200 mL 1 x SDS Running Buffer and 500 μL NuPAGE Antioxidant to the electrophoresis tank and mix well. Add 600 mL 1 x SDS Running Buffer to the lower tank. Add 7 μL ~ 10 μL protei...

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Abstract

The invention discloses a babesia caballi disease immunoblotting detection method and a kit. The babesia caballi disease immunoblotting detection method is established on the basis of obtaining a diagnostic antigen and positive control serum through a goat anti horse second antibody compound marked by horse radish peroxidase and a chemiluminescence substrate. The method has the advantages of less reagent dosage, low cost, high specificity, high sensitivity, quicker operation, no toxicity and harm, low required condition, easy judgment of a test result, long-term storage and stronger specificity than ELISA (Enzyme-Linked Immuno Sorbent Assay). By detecting by using the method, false positive and false negative results are not detected.

Description

Technical field [0001] The present invention involves a method of immunohistocytics and kit preparing methods of Baberz. It specially involves a method of using an immunohistic test test to detect Babesia Caballi (Babesia Caballi) serum antibody and its diagnostic kitIt belongs to the field of raw horseshoma of horses. Background technique [0002] Babesia Caballi (Babesia Caballi) is a hemorrhoid disease caused by the red blood cells in the red blood cells of the macanols in the horse. The clinical manifestations are symptoms such as high fever, anemia, jaundice, and edema.The most acute cases are relatively rare, and the diseased horses have died or died when they are discovered.The disease is widely distributed in the world. In Europe, France, Italy, and southern and most parts of the eastern parts, it is reported by Baberz insect disease in 58 degrees north latitude.The epidemic survey of Baberz Worms in southern France was 11.3%, and there were reports of Baberz worm disease...

Claims

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Application Information

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IPC IPC(8): G01N33/569C12N1/10
Inventor 王玉玲左锋王建华柴宏森王乃福赵林立侯艳梅赵祥平
Owner 中华人民共和国天津出入境检验检疫局
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