Discovered back in 1942 by Koons, the immunofluorescence reaction is not a new research method. However, the emergence of hybridoma technologies that allowed the production of monoclonal antibodies, gave a "second life" to this reaction, since their use allowed several times to increase the sensitivity of this reaction and its specificity.
And today we will tell you in detail about the reaction of direct and indirect immunofluorescence( RIF) as a method for diagnosing Koons for adult men and women during pregnancy.
What is the immunofluorescence reaction
Presenting an excellent opportunity for rapid obtaining of an accurate diagnosis, the immunofluorescence reaction makes it possible to determine the presence of a pathogen in a pathological material. For this, a smear is used from a material that is specially treated with a labeled FITC( fluorescein isothiocyanate), and is studied as a heterogeneous assay.
To obtain the result, a fluorescent microscope is used, in its optical system there is a set of light filters to provide the preparation with blue-violet or ultraviolet light having a predetermined wavelength. This condition allows the fluorochrome to glow for a given range. The researcher evaluates the properties of luminescence, its nature, the size of objects and their mutual location.
To whom it is prescribed
Immunofluorescence may be administered in the diagnosis of a variety of viral diseases. In particular, it is prescribed in a comprehensive examination to identify the following factors:
- the presence of the papilloma virus in the body;
- infection with salmonella;
- existence in the body of certain antigens;
- reveals the probability of infection of the body with chlamydia, mycoplasmas and other microorganisms that have the ability to excite viral diseases of man;
- diagnosis of viral diseases in animals.
The above indications allow the use of the immunofluorescence reaction in the detection of viral diseases of various nature in humans and animals.
Objectives of the
Because this diagnostic method has many advantages, including its high effectiveness, speed and results, as well as the absence of a large number of contraindications, it determines the presence of viral infections in the body. Therefore, this analysis can be assigned both for setting and for clarifying the diagnosis on the basis of which a treatment regimen is prescribed.
Carrying out the procedure does not cause unpleasant sensations, for it it is necessary to obtain material for analysis, which is taken from any body fluid: saliva, sputum, scraping from the surface of the mucous membranes. Blood can also be taken for analysis. The frequency of carrying out the reaction of immunofluorescence is assigned by the attending physician, who needs to obtain data on the dynamics of the processes occurring in the body.
Types of this procedure
Today, several versions of this analysis are used, each of which has a number of specific features and allows you to get the most detailed picture of the processes occurring in the body.
To varieties of the reaction of immunofluorescence should be attributed:
- enzyme-linked immunosorbent assay is one of the most rapidly developing types of diagnostics, this analysis enables the production of quantitative data without the use of serial dilutions. Using the obtained measurements of the optical density of the liquid, it is possible to accurately determine the concentration level of the desired component. The wide possibilities of this type of analysis are used when using monoclonal antibodies for its implementation, which allows to determine the phase of the infectious process, its severity;
- DNA Diagnostics - this method is based on complementary binding of nucleotides, for which liquids such as saliva, blood, liquor, urine, sputum, biopsies, blood can be used. This method most effectively reveals the presence of papilloma viruses in the body, however many modern test systems can occasionally give false positive and false negative results. They can be caused by contamination of fluid samples for the analysis of specific DNA, the presence of which can be nested or total;
- immunochromotography - the specificity of this method for determining the presence of a pathological environment and viruses in an organism consists in the use of labeled antibodies during the reaction. This diagnostic method is used to identify and measure the activity of the group A streptococcal infection process, as well as chlamydia of the following species: ClamikitR Innotech International, ClearviewTM Chlamydia from Oxoid. Possessing the highest sensitivity, test-systems, which are based on this method of investigation.usually apply as an orientation test.
The listed varieties have specific features and specific characteristics of the results, but all of them are aimed at obtaining data on the presence of pathological microorganisms and viruses in the organism, as well as the degree of their multiplication and activity.
Indications for
The immunofluorescence reaction can be assigned to detect any pathological environment in the body.
Chlamydia, trichomonads, gonococci and streptococci, as well as lamblia of all types are determined when carrying out this type of diagnosis. Bullous dermatitis and dermatitis Duhringa, herpes on the lip and other diseases also require a RIF. The appointment of a physician for its implementation is mandatory.
Contraindications for carrying out
Since any kind of body fluid is required as the test material for carrying out this reaction, taking them usually does not constitute difficulties and there are no contraindications for carrying out the immunofluorescence reaction. However, in pregnancy and in children up to 6 months, taking the material for the study is carried out with the utmost caution.
The absence of contraindications allows carrying out this type of diagnosis when appointing a doctor to all patients. Its safety is guaranteed by the use of disinfected tools and disposable syringes.
Preparing for the
procedure There are no features of taking material for this analysis. Blood is taken for him on an empty stomach so that there is not a high content of substances in it that can change the true readings and give a false picture.
. As the sampling of the
assays is carried out. Since no special preparation is required for the analysis, only food intake is excluded 12 hours before the test and no medicines are taken, the material is taken as a normal process of taking body fluids for analysis.
Subjective sensations during the procedure may vary depending on the sensitivity.
Decoding of results
Application of modern test systems allows obtaining the most accurate results of analysis. The following data are used to decipher the result:
- fluorescence intensity degree;
- shade of fluorescence;
- peripheral character of the luminescence process of the object;
- characteristics of morphology, the location of the pathogen in the taken smear of the test material and its dimensions.
During the examination of objects that are large in size( for example, gardenerellas, trichomonads, cells that are already infected with viruses), the criteria listed above make it possible to obtain the most reliable results. However, the elementary bodies of mycoplasma and chlamydia have dimensions lying at the limit of the resolving powers of the luminescent microscope, which makes it difficult for the
to obtain an accurate result, since the peripheral luminescence loses some of its intensity. The remaining criteria are already insufficient to accurately identify the microorganisms under investigation. For this reason, special requirements are imposed on the specialists who conduct this type of research: the level of their qualification should be sufficient to operate with the available data.
For this reason, only the physician with the appropriate skill level can decipher the analysis obtained. About the price of research using the RIF method, read below.
Average cost of
The price of carrying out the immunofluorescence reaction depends on the place of its carrying out and the level of the medical institution, as well as the qualifications of the conducting expert. Today the cost ranges from 1280 to 2 160 rubles.
More details about immunological reactions will be revealed by the video below: