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Detection kit for enterococcus faecium indirect blood clotting and application thereof

A detection kit and technology for Enterococcus faecium, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as lack of serological diagnostic methods, loss of animal husbandry production, misdiagnosis and missed diagnosis of Enterococcus faecium infection, etc., to achieve Easy to store for a long time, low price, avoid the effect of short storage time

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 Enterococcus faecium infection at the grassroots level, thus causing unnecessary losses to animal husbandry production

Method used

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  • Detection kit for enterococcus faecium indirect blood clotting and application thereof
  • Detection kit for enterococcus faecium indirect blood clotting and application thereof
  • Detection kit for enterococcus faecium indirect blood clotting and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation of Enterococcus faecium indirect hemagglutination diagnostic kit

[0027] The components of the Enterococcus faecium indirect hemagglutination diagnostic kit are as follows:

[0028] (1) 1 bottle of Enterococcus faecium 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 Enterococcus faecium 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 high pressure. ...

Embodiment 2

[0058] Apply the test kit of the present invention to detect the serum to be tested, the specific method is as follows:

[0059] 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 (that is, Enterococcus faecium indirect hemagglutination 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 place it in a 37°C incubator for 1.5 -2h, judgment result.

[0060] Judgment criteria: all erythrocytes agglutination (++++); 75% erythrocyte aggl...

Embodiment 3

[0061] Example 3 Specificity experiment of the Enterococcus faecium indirect hemagglutination detection kit of the present invention

[0062] Use the Enterococcus faecium indirect hemagglutination detection kit of the present invention to detect healthy rabbit serum, streptococcus positive serum, Staphylococcus positive serum, Enterococcus faecalis positive serum and Enterococcus faecium, the results except for Enterococcus faecium, the rest are all Negative (see Table 2), indicating that the antigen has a very high specificity.

[0063] Table 2 Specific test results of Enterococcus faecium indirect hemagglutination diagnostic reagents

[0064]

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Abstract

The invention provides a detection kit for enterococcus faecium indirect blood clotting. The detection kit is characterized by comprising an enterococcus faecium indirect blood clotting antigen, wherein the enterococcus faecium indirect blood clotting antigen is composed of enterococcus faecium CCTCC M 2016047 whole bacterial proteins and hydroformylation-tannin sheep red blood cells. The kit provided by the invention is simple, convenient and quick in operation; special equipment and instrument are not required; the whole process can be completed within 2-3 hours; the detection kit is fit for popularization and application in basic level; the detection kit can be widely applied to the investigation of enterococcus faecium epidemiology; the repeatability of the result is excellent; the result is stable and reliable; the antibody of the enterococcus faecium 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 detection kit for indirect hemagglutination of Enterococcus faecium. Background technique [0002] Enterococcus Faecium is a Gram-positive, catalase-negative coccus that is the main flora in the intestinal tract of humans and animals. Since the 1990s, Enterococcus faecium has gradually become an important conditional pathogen of humans and animals. At present, in terms of animal research, E. faecium infection mainly relies on laboratory bacterial isolation and PCR molecular biology identification. At the grassroots level, there is a lack of rapid and effective serological diagnostic methods for E. faecium. The lack of serological diagnostic methods may cause misdiagnosis and missed diagnosis of Enterococcus faecium infection at the grassroots level, thus causing unnecessary losses to animal husbandry production. Contents of the inven...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/56944G01N33/68
Inventor 刘永安李学瑞王立燕马丽娜周建华刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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