Ureaplasma fluorogen

Diagnosticum for detecting Ureaplasma urealyticum antigens in the immunofluorescence reaction

The diagnosticum is designed to detect the direct immunofluorescence reaction of Ureaplasma urealyticum in smear preparations from clinical materials obtained from patients with inflammatory diseases of the urogenital tract.

COMPOSITION OF THE SET

FITC antibodies to Ureaplasma urealyticum - lyophilized rabbit specific polyclonal antibodies labeled with fluorescein isothiocyanate (FITC) containing Evans blue and a preservative (sodium azide); loose amorphous mass of blue.

Antibody Dilution Solution (PPA) - phosphate-buffered saline containing a preservative (sodium azide); clear colorless liquid.

The set also includes glass slides decorated with 10 holes.

OPERATING PRINCIPLE

In the presence of the corresponding pathogen in the test sample, FITC-labeled antibodies specifically bind to its antigens, which is detected by luminescence microscopy due to the intense green glow of these complexes, which contrasts with the red color of the epithelial cells stained with Evans blue.

 RESEARCH SAMPLES

Smear preparations prepared from the separated urethral mucosa, vagina, cervical canal, prostate secrets, ejaculates, cell sediment from centrifuged urine samples are subject to investigation for the presence of Ureaplasma urealyticum.

Apply the test material with a thin layer to the surface of a clean fat-free glass slide, air dry and fix in ethanol (96%) for 15 minutes, then dry it again in air. The drug should contain as many epithelial cells as possible.

Shelf life of fixed smear preparations at 4 ºC is no more than three days. Longer storage (up to 1 month) is allowed only at minus 70 ° C, repeated freezing-thawing of drugs is not allowed.

 ANALYTICAL AND DIAGNOSTIC CHARACTERISTICS

Diagnostic sensitivity and diagnostic specificity of the kit-100%

The basic kit is designed to study 50 samples, including control. At the request of the consumer, the basic set of the kit (the number of individual packages with reagents and their volumes) can be changed.

 MODE OF APPLICATION

Equipment and materials

The chamber is wet for incubation at a temperature of 37 C.

Luminescent microscope with light filters FS 1-2, BS 8-2, SZS 7-2, ZhS-18.

Immersion oil (non-fluorescent).

Water is tap.

Purified water.

Rectified ethyl alcohol (96%).

Preparation of reagents and materials

To withstand all components of the kit at a temperature of 18 to 25 C for at least 30 minutes.

In the vial with FITZ antibodies, add the contents of the vial with PPA, dissolve for 3-5 minutes with stirring.

Store the antibody solution at a temperature of 2 to 8 ° C in a dark place for no more than 14 days.

If necessary, longer storage (up to 1 month). Pour the antibody solution into small portions and store at a temperature of minus 20 C.

Analysis

1. Apply 30 μl of antibody solution to a fixed smear preparation and incubate for 20 min in a humid chamber at 37 ° C.

Attention! It is not allowed to dry the drug with a solution of antibodies.

2. Rinse smear with tap water for 15 minutes, rinse with distilled water and air dry at room temperature.

Attention! A thorough washing of the drug is necessary to eliminate the non-specific glow during microscopy, the source of which is the presence of a large amount of mucus in the test material.

3. To investigate the smear preparation under a luminescent microscope with a set of light filters: FS 1-2, BS 8-2, SZS 7-2, ZhS-18 and with an increase of 600-1000x.

For microscopy, use oil immersion or a water-immersion system.

Attention! When using oil immersion, only special (non-fluorescent) immersion oil should be used.

 Profitability Analysis

When luminescent microscopy, ureaplasmas are detected in the form of an intense green granular glow on the membranes of epithelial cells stained in reddish color and / or in the intercellular space.

The result is considered positive if 10 or more bright green granules are clearly identified in the whole preparation, which clearly stand out against the reddish background of the preparation.

In the absence of a specific (intense green) glow in the preparation, the result is considered negative.

If there are no epithelial cells in the preparation or less than 10 bright green granules are found, the study must be repeated, paying particular attention to the correct sampling of the test material.

Yellow or greenish yellow fluorescence under microscopy is considered an artifact. In this case, the study also needs to be repeated.

ISSUE FORM

Available in the kit:

FITZ antibodies to Ureaplasma urealyticum 0.25 ml - 1 bottle.

PPA 1.5 ml - 1 bottle.

Slides - 5 pcs.

Each set