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Staphylococcus Warner indirect blood coagulation detection kit and application thereof

A detection kit and Staphylococcus detection technology, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as misdiagnosis and missed diagnosis of Staphylococcus warnerii infection, loss of animal husbandry production, lack of serological diagnostic methods, etc., to achieve Easy to store for a long time, cheap, easy to operate

Active Publication Date: 2016-08-17
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The lack of serological diagnostic methods may cause misdiagnosis and missed diagnosis of Staphylococcus warnerii infection at the grassroots level, thus causing unnecessary losses to animal husbandry production

Method used

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  • Staphylococcus Warner indirect blood coagulation detection kit and application thereof
  • Staphylococcus Warner indirect blood coagulation detection kit and application thereof
  • Staphylococcus Warner indirect blood coagulation detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation of Staphylococcus warnerii Indirect Hemagglutination Diagnostic Kit

[0027] The components of the Staphylococcus warnerii Indirect Hemagglutination Diagnostic Kit are as follows:

[0028] (1) 1 bottle of Staphylococcus warnerii indirect hemagglutination antigen, 10ml / bottle;

[0029] (2) 1 bottle of standard positive serum, 1ml / bottle;

[0030] (3) 1 bottle of standard negative serum, 1ml / bottle;

[0031] (4) 2 bottles of diluent, 10ml / bottle.

[0032] The shelf life of the kit is one year.

[0033] 1 Preparation of Staphylococcus warnerii indirect hemagglutination antigen

[0034] 1.1 Culture medium preparation

[0035] Prepare THB liquid medium 2000ml: 4.0g beef powder, 40g tryptone, 40g glucose, 6.0g sodium chloride, Na 2 HPO 4 5.8g, KH 2 PO 4 Add 0.6g, Yeast extracts 6.0g to deionized water, set the volume to 2000mL, fully stir to dissolve, and use anhydrous Na 2 CO 3 Adjust the pH to 7.4. Sterilize under high temperature and hig...

Embodiment 2

[0057] Apply the test kit of the present invention to detect the antigen to be tested, the specific method is as follows:

[0058] First add 75 μL of diluent to each well of 8 rows and 12 columns of a 96-well “∨” polystyrene titration plate; In column 1, add 25 μL of standard positive serum and standard negative serum to the first well of columns 10 and 11, respectively, and column 12 is a blank control. Use a row gun to thoroughly mix wells 1-11, then pipette 25 μL into well 2, serially dilute to well 8, discard 25 μL from well 8 after mixing; finally add 1% sensitized red blood cells to each well Suspension (i.e. Staphylococcus warnerii indirect hemagglutination diagnostic antigen with a concentration of 100 μg / ml) 25 μL, after adding the sample, place the ∨-shaped titer plate on a micro shaker for 1-2 minutes, cover with a glass plate, and set the temperature at 37 °C Box 1.5-2h, judge the result.

[0059] Judgment criteria: all erythrocytes agglutination (++++); 75% eryt...

Embodiment 3

[0060] Example 3 Specificity test of the Staphylococcus warnerii indirect hemagglutination detection kit of the present invention

[0061] With the Staphylococcus warnerii indirect hemagglutination detection kit of the present invention, healthy rabbit serum, Staphylococcus warnerii, Streptococcus positive serum, Staphylococcus positive serum, Enterococcus faecalis positive serum, Enterococcus faecium positive serum are detected, the result Except for Staphylococcus warnerii, the rest were negative (see Table 2), indicating that the antigen has a very high specificity.

[0062] Table 2 Specific test results of indirect hemagglutination diagnostic reagents for Staphylococcus warnerii

[0063]

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Abstract

The invention provides a staphylococcus Warner indirect blood coagulation detection kit and application thereof. The kit is characterized by comprising a staphylococcus Warner indirect blood coagulation antigen, wherein the staphylococcus Warner indirect blood coagulation antigen is composed of whole bacterial protein of staphylococcus Warner CCTCC M2016048 and hydroformylated-tanned sheep red blood cells. The kit provided by the invention is simple to operate and is convenient and rapid, and special equipment and instruments are not needed; and a whole process is finished within 2-3 hours, so that the kit is suitable for being popularized and applied to a basic level and can be widely applied to investigation and study of staphylococcus Warner disease epidemiology. A detection result has good stability and is stable and reliable, and an antibody of staphylococcus Warner can be accurately detected.

Description

technical field [0001] The invention belongs to the technical field of detection reagents for microorganisms, and in particular relates to a staphylococcus warnerii indirect hemagglutination detection kit and application thereof. Background technique [0002] Staphylococcus warnerii is a Gram-positive, catalase-negative coccus that is the main flora in the intestinal tract of humans and animals. Since the 1990s, Staphylococcus warnerii has gradually become an important opportunistic pathogenic bacteria for humans and animals. At present, Staphylococcus warnerii infection mainly relies on laboratory bacterial isolation and PCR molecular biology identification, but there is a lack of rapid and effective serological diagnostic methods for Staphylococcus warnerii at the grassroots level. The lack of serological diagnostic methods may cause misdiagnosis and missed diagnosis of Staphylococcus warnerii infection at the grassroots level, thus causing unnecessary losses to animal hu...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/56938G01N33/68
Inventor 马丽娜李学瑞周建华刘永安王立燕刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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