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RECOMMENDATIONS FOR PRACTICAL WORK

Task 1. When you do the venereal disease research laboratory (VDRL) you must put 0.1-0.2 ml the serum from patients in the tubes, then add carefully the dilution of the antigen. If you do all the correct, the measure between serum and antigen will be clear. The tubes are incubated by 37°Ñ for 30 minutes. In work time, you must be careful because the serum from patients may contain spirochetes. If the result will be positive, white ring will be at the measure between serum and antigen.

Task 2. The Wassermann’s test with patient’s sera.

N tubes, contain 1 (Test) 2 (Control of the serum) 3 (Control of the antigen)
Serum from patients 0,5 0,5 --
Àantigen 0,5 -- 0,5
Complement 0,5 0,5 0,5
0,9 % NaCl -- 0,5 0,5
37°Ñ, 30 minutes
Hemolytic mixed 1,0 1,0 1,0
37°Ñ, 60 minutes
Result ? -- --

Task 3.

T. pallidum is in IFT. Tissue fluid or exudate is spread on a glass slide, air dried, and sent to the laboratory. It is fixing, staining with a fluorescein-labeled antitreponeme serum, and exam­ined by means of immunofluorescence microscopy for typical fluorescent spirochetes.

T. pallidum is a slender spiral measuring about 0.2 μm in width and 5-15 μm in length; it has 10-12 wave curls and actively moves. It will green colore shining in IFT test.

 

 

T. pallidum is in the The Dark field examination.1-2 drops of tissue fluid or exudates is placed on a slide and cover slip pressed over it to make a thin layer. The preparation then examined under oil immersion with dark field illumination for typical motile spirochetes.

 

 

 

T. pallidum. Silver impregnation method.

 

Borelia persica stained by Giemsa’s are purple irregular spirals 10-30 μm long and 0.3 μm wide. The distance between turns varies from 2 to 4 μm.

 

 

 

Borelia persica in the dark field.

 

 

 

Leptospira interrogans in the dark field. It does not stain with aniline dyes. Leptospirae have 18-20 prime waves and two secondary (one of the each ends).

 

 

Task 4.

Cardiolipin antigen for Wassermann’s test is extract of lipids from meat hart of bull with addition of 0.25-0.3% cholesterin and lecithin. It has permanent composition and does the standard resultants.

Specific antigen for Wassermann’s test is making from spirochetes, which are growing in the rabbit testicles and are destroyed by the supersonic.

Cardiolipin antigen for venereal disease research laboratory on the glass is a solution of the cardiolipin, cholesterin and lecithin.

Wassermann’s test is because regain-containing sera fix complement in the presence of cardiolipin antigen. If the test is positive, we will see the sediment of the erythrocytes at the bottom of the tube.

Killed leptospirosis vaccine contains killed by heat and conserved by phenol leptospires. It is using by epidemiological indications.



Addition 1


Date: 2016-01-14; view: 267


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Laboratory diagnostics of the anthrax and brucellosis | Microbiology of the Spirochetosis (the syphilis, borreliosis and leptospirosis).
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