Methods for detection of Leishmania in smear from the skin infiltration

To identify Leishmania examined fixed and stained smear contents infiltration.

Reagents

• Methyl alcohol, or absolute ethyl alcohol, or a mixture of Nikiforov (a mixture of equal amounts of sulfur ether and absolute ethanol).
• Phosphate buffer pH 6.9–7.1, for the preparation of which is mixed 40 ml KH₂RO₄ (9,078 g dry matter KH₂RO₄ in 1 liter of solution) and 60 ml Na₂PHO₄ (11,876 g dry matter 1 liter of solution).
• Azureozin Romanovsky.

Methods of detection of Leishmania in smear from the skin infiltration

Material for the study taken from tubercle (at an early stage of the process) or from the regional infiltration ulcers (at a later stage), receded somewhat from its edge.

Rubbing alcohol with cotton bump or selected area of ​​infiltration, squeeze it between two fingers to bleeding and, without weakening the pressure of fingers, the end of a sharp scalpel (it is desirable to the eye) make the surface of the skin incision. When the blood should remove gauze. From the bottom and edges of the incision is made with the same scalpel scraping the affected tissue quickly and smear the contents of scraping a thin layer on a slide.

Prepare a few strokes. Smears were dried in air, fixed and stained with Romanovsky.

Evaluation of the results of detection of Leishmania in smear from the skin infiltration

The preparation should be clearly distinguishable macrophages, endothelial, plasma cells and lymphoid, fibroblasts and a small admixture of peripheral blood cells.

The presence in the preparation of epithelial cells, as well as unstructured clumps, painted uniformly lilac, means, that the scraping was taken too superficial and should be repeated. The smear should not be too much blood, and pus and bacteria.

Leishmania L. tropica found in macrophages, as well as outside the cells in the form of round, oval or elongated cells measuring 3 5 m. When stained with Romanovsky cytoplasm of Leishmania gets light blue or purple color, the nucleus becomes red-purple.

Clearly visible kinetoplasts, intensely colored, than the core.

In the initial stages of Leishmania ulceration are easily detected in the bumps and sores in the regional infiltration. In purulent sores can only be found deformed and destroy Leishmania, which are difficult to differentiate. At the stage of healing in the affected tissues Leishmania is rarely detected.