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A kind of kit for detection of enterococcus faecalis indirect hemagglutination and its application

A detection kit and technology for Enterococcus faecalis, applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of misdiagnosis and missed diagnosis of Enterococcus faecalis infection, loss of animal husbandry production, lack of serological diagnostic methods, etc. Easy to store for a long time, inexpensive, avoids the effect of short storage time

Active Publication Date: 2017-12-12
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 faecalis infection at the grassroots level, thus causing unnecessary losses to animal husbandry production

Method used

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  • A kind of kit for detection of enterococcus faecalis indirect hemagglutination and its application
  • A kind of kit for detection of enterococcus faecalis indirect hemagglutination and its application
  • A kind of kit for detection of enterococcus faecalis indirect hemagglutination and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation of Enterococcus faecalis Indirect Hemagglutination Diagnostic Kit

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

[0028] (1) 1 bottle of Enterococcus faecalis 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 faecalis 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 pressu...

Embodiment 2

[0058] Apply the test kit of the present invention to detect the antigen 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 faecalis indirect haemagglutination diagnostic antigen with a concentration of 40 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 1.5-2h, judge the result.

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

Embodiment 3

[0061] Example 3 Specificity test of the Enterococcus faecalis indirect hemagglutination detection kit of the present invention

[0062] Use the Enterococcus faecalis indirect hemagglutination detection kit of the present invention to detect healthy rabbit serum, Enterococcus faecalis positive serum, Streptococcus positive serum, Staphylococcus positive serum, and Enterococcus faecium positive serum, and the results except Enterococcus faecalis, The rest were negative (see Table 1), indicating that the antigen has a very high specificity.

[0063] Table 2 The results of the specificity test of Enterococcus faecalis indirect hemagglutination diagnostic reagents

[0064]

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Abstract

The invention provides an enterococcus faecalis indirect hemagglutination detection kit which is characterized by comprising an enterococcus faecalis indirect hemagglutination antigen. The enterococcus faecalis indirect hemagglutination antigen is composed of the whole bacterial proteins of enterococcus faecalis CCTCC M 2016027 and hydroformylation-tanned sheep red blood cells. The enterococcus faecalis indirect hemagglutination detection kit has the following advantages: the operation is simple, convenient and quick; special equipment and instruments are not required; the entire operation is completed in 2-3 hours. The enterococcus faecalis indirect hemagglutination detection kit is suitable for promotion and application in grassroots institutions, and can be extensively applied in the research and study on enterococcus faecalis epidemiology. The enterococcus faecalis indirect hemagglutination detection kit delivers an excellently repeatable, stable and reliable result, and can accurately detect the antibody of enterococcus faecalis.

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 enterococcus faecalis indirect hemagglutination and an application thereof. Background technique [0002] Enterococcus faecalis ( Enterococcus faecalis ) is a Gram-positive, hydrogen peroxide-negative coccus, one of the main flora in the intestinal tract of humans and animals. Since the 1980s, Enterococcus faecalis has gradually become an important conditional pathogen of humans and animals, which can cause bacteremia, urinary tract infection, endocarditis and other diseases. At present, in terms of animal disease diagnosis, E. faecalis 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. faecalis. The lack of serological diagnostic methods may cause misdiagnosis and mis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/56916G01N33/56944G01N33/68G01N2333/315
Inventor 刘永生李学瑞周建华马丽娜刘永安王立燕
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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