Serum Legionella, 2 ml, DENKA SEIKEN, Japan

Legionella is a gram-negative bacterium that is mobile because it has flagella and causes legionella.

Serotypes used for serotyping of Legionella are liquid products with type specific agglutinins.

When using this product, the group is identified by agglutination on the glass.

Serums are prepared by hyperimmunization of rabbits to reference strains of ATCC. For immunization, strains are mixed with B-CYE (Buffer Coal and Dried Extract) and inactivated by heating up to 100 ° C. After blood collection, the sera are separated, inactivated by heating to 56 ° C. for 30 minutes, and the nonspecific agglutinins are removed by absorption and sterilized by filtration. Use 0.08% sodium azide as a preservative.

PRODUCT DESCRIPTION

Legionella serums are produced from the blood of rabbits and contain 0.08% sodium azide as a preservative. Relevant serotypes are packed in vials of 2 ml with droppers and ready for use.

1) L.pneumophilia Serogroup 1

2) L.pneumophilia Serogroup 2

3) L.pneumophilia Serogroup 3

4) L.pneumophilia Serogroup 4

5) L.pneumophilia Serogroup 5

6) L.pneumophilia Serogroup 6

7) L. Boszemanii

8) L.dumoffi

9) Gormani

10) L. micdadei

SCOPE OF APPLICATION

Determination of Legionella serogroups.

PRINCIPLE OF ACTION

When mixed with the corresponding serums containing antibodies to the Legionella strains in the serum of the patient, agglutinate is formed. The reaction is macroscopic and allows you to identify each serotype.

PROCEDURE FOR IMPLEMENTATION

1. The material is needed to perform the analysis that is missing from the set.

Object glasses, glass pencil, pipettes and micropeak, microbiological loop, physiological solution, autoclave (121ºC), water bath (100ºC), centrifuge.

2. Preparation of reagents

Whey is ready for use.

3. Sample

The microbiological culture is derived from pure culture and identified as Legionella by biochemical tests and should be serotyped. If the sample consists of a large number of strains, the serotypes must be properly identified.

1. Procedure

1) Dissolve a certain amount of bacterial mass (3-5 times a microbiological loop (match head) in 3 ml of physiological solution and warm at 121 ° C for 15 minutes or at 100 ° C. for 1 hour. Centrifuge at 900 revolutions for 20 minutes, remove the surface layer, dissolve the precipitate at 0 , 5 ml of saline solution and used as a suspension of antigen

2) Provide a drop of each serum and physiological solution-like control (30μl) on a clean, substantive glass of it to separate particles using a pencil to write on the glass

3) Provide a suspension of antigen (5-10μl) in the serum and a physiological solution on the lens.

4) Mix the reagents by careful movement of the glass back and forth for 1 minute and observe the formation of particles of agglutinates. Agglutination is very noticeable if the glass is viewed through a stream of fluorescent light. The primary evidence of agglutination confirmation is its absence between the suspension of the antigen and the physiological solution. If within 1 minute there is pronounced agglutination, the reaction is considered positive. Prolonged in time or poor agglutination is considered negative.

WARNINGS IN THE INTERPRETATION OF RESULTS

1. Annealing on the glass, which lasts more than 1 minute after the reaction between the anti-gene and the antibody, is evaluated as negative.

2.If there is positive agglutination, then a certain antigen having specific properties.

3. A cross-reaction between two or more serums, requiring warming up of a bacterial suspension for repeated testing

4. For serogroups 4 and 5, a complex serotype may be isolated in rare cases.

5. When agglutination is present in the reaction of the antigenic suspension and the physiological solution, the test must be performed again by isolating another colony.

TESTING

1. Sensitivity test

1) When one drop of a product reacts to a reference glass with a reference strain of known serotype, granular agglutination is very noticeable.

2. Specificity of the test:

The test is carried out by a simple method for determining the sensitivity of the test, when the serum is agglutinated only with the reference strain, and in response to an inappropriate reference, macroscopic agglutination is not observed.

Delivery time: 5 days

Information updated: 02.08.2018