Method of obtaining standard quality sample of saliva facia for crystallography

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to clinical, laboratory diagnostics, microbiological methods of research, and is aimed at standardisation of saliva analysis by method of wedge dehydration/crystallography. Claimed is method of obtaining standard, quality sample of mixed saliva facia for crystallography with application of portable laboratory device with inbuilt levels on axes X and Y, legs, regulated by height and isolating cover. From 20 to 40 microscope slides are simultaneously placed on the surface of portable laboratory device after obtaining smooth horizontal without inclination angle surface. After that, one sample of mixed saliva in amount 0.02 ml, preliminarily centrifuged for 20 min at 3000 rev/min, is applied on each microscope slide by means of micropipette. As a result a round 1.0 mm high drop of mixed saliva with diameter from 4.0 to 5.0 mm, corresponding to standard parameters, is obtained. Then, standard in dimensions drop on microscope slide placed on the surface of portable laboratory device, adjusted by levels, is dried at room temperature +18…+25°C under isolating cover for 5 hours, with further performance of microscopy of standard quality sample of saliva facia.

EFFECT: obtained sample makes it possible to interpret indices of crystallography without distortions, as standard in volume, dimensions and shape drop of mixed saliva is obtained, and there is no displacement of crystallisation centre and impairment of figures of saliva facia (crystallography pattern) in drop, dried on ideal horizontal surface of the device, ratio of central and peripheral zones of facia exactly correspond to organism's condition, which reduces percentage of false results of crystallography.

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The invention relates to medicine, namely clinical laboratory diagnosis, microbiological methods, and aims at standardizing research saliva method wedge dehydration/crystallography.

Study of physico-chemical and metabolic parameters of saliva available biological fluids non-invasive methods, is becoming increasingly important. Currently increasingly important method wedge dehydration/crystallography saliva, which was opened by the researcher that is, rapes. Russian scientists Century. N. Shabalin and S. N. Shatokhina revealed the main regularities of the formation of structures of the solid phase bioliquids, classified and given an objective description of their system and local morphological features (barer, M., Denisov A. B., Evocative I. P., I. Mikhaleva N. / Crystallization oral fluid. The composition and frequency of the substrate surface //. Bulletin of experimental biology and medicine, 2007. - 12. - S. 693-696). Implementation of the method is that when drying a drop of biological fluid is formed dry film or facies, representing structural macropore dry residue, which is clearly distinguished by the shape, orientation, size, and other parameters figure facies (solid residue of saliva). The importance of the first factors determine the nature of structure formation of saliva, water and the total concentration of dissolved mineral and organic substances, as well as their relationship.

There is a way wedge dehydration/crystallography mixed with saliva, developed by the Russian scientists Century. N. Shabalina and S. N. Athinai (, Moscow 2001).

But the disadvantages of the prototype are:

- applying drops of investigated liquid on a glass slide uncertain size, making it difficult to identify and leads to bias data;

- drying drops of the liquid under study on the slide is held in thermostat, which leads to a rapid, uneven drying the droplets to the distortion figures facies saliva and, as a result, false result;

- no thermostat flat horizontal surface does not allow the drop evenly to dry, which leads to distortion ratio of the Central and peripheral zones, facies - the main parameter of the study;

- hit probability in a drying drop of foreign particles due to the absence of a protective insulating coating causes artifacts facies of saliva.

The goal is to get a quality standard sample facies mixed with saliva for studies wedge dehydration / crystallography.

The technical result consists in the fact that mo is to get a quality standard facies mixed with saliva for crystallography uses a portable laboratory with built-in levels on the X and Y axes, height-adjustable legs and an insulating cover.

The technical result is achieved by the fact that at the stage of preparation of saliva sample for deposition on a glass slide and further drying is used, the device is portable laboratory with an insulating cover. Pre-aligned horizontal surface of the device is portable laboratory using the adjustable legs under the control of the built-in surface levels on the X-axis and y After receiving a flat horizontal without error angle of inclination of the plane on the surface of the device portable laboratory are put slides from 20 to 40 pieces at a time. Then with the help of micropipette "Inpipe" on each glass slide is applied to one sample of mixed saliva in the amount of 0.02 ml, previously centrifuged for 20 min at 3000 rpm In the relevant standard settings drop mixed with saliva: height drops of 1.0 mm diameter drops from 4.0 to 5.0 mm, the shape of the drop rounded without distorting the form. Next standard size drop on a glass slide, placed on the surface of the portable device laboratory, configured, dried at room temperature +18...+25°C under an insulating lid for 5 hours, then is he who microscopy standard quality sample facies saliva. Thus, the resulting model allows us to interpret the indicators of crystallography without distortion, because the obtained standard volume, size and shape of a drop mixed with saliva, and drying the droplets is carried out on a perfect horizontal surface of the device, resulting in no displacement of the centre of crystallization, the violation of figures facies saliva (figure crystallography), and the ratio of the Central and peripheral zones facies correspond exactly to the condition of the body, which reduces the percentage of false positives crystallography.

Used portable laboratory is shown in Fig.1, where the index 1 - shelf extruded acrylic glass, the pointer 2 levels on the X-axis and Y-index 3 - adjustable legs, index 4 - slides, index 5 - insulating cover, which prevents the influence of the external environment and maintains a constant humidity and temperature, 6 - hole insulating cover.

Inside portable laboratory has two levels, located at right angles relative to each other. Using layers, you can estimate the inclination of the surface relative to the horizon along the X-axis and y-axis When placing the level on the X-axis and Y-axis bubble inside should be placed in a Central position. The scheme of obtaining high-quality with andariego sample facies saliva for crystallography is shown in Fig.2, where a is the standard sample facies mixed with saliva.

After fixing the parameters necessary to obtain a flat surface using a portable laboratory device on a glass slide by using the pipettor is applied to 0.02 ml of mixed saliva. Get a drop of mixed saliva with standard parameters: height drops of 1.0 mm, diameter 5.0 mm, the shape of the drop rounded without distorting the form, and when it is drying, get a sample of facies saliva without displacement of the center of crystallization and without breaking figures facies, the ratio of the Central and peripheral zones corresponds to the condition of the body.

In the case when the surface alignment is not carried out through a level, get a drop of mixed saliva with custom parameters: height drops of 1.0 mm, a diameter of 10.0 mm, the shape of the drop is deformed edge, distorted in shape, and when it is drying receive a custom sample facies saliva with offset center of crystallization and a violation of the texture picture facies without the ability to determine the ratio of the Central and peripheral zones, which results in a false result in the diagnosis of that shown in Fig.3, where B is a distorted sample facies of saliva, which is impossible to investigate the method of crystallography.

The advantage of the proposed method consists in the following:

<> - leveling horizontal surface by means of additional equipment levels on the X and Y axes in the portable laboratory allows to get a little bit mixed with saliva, the corresponding standard parameters: height drops of 1.0 mm, a diameter of from 4.0 to 5.0 mm, the shape of the drop rounded without distortion, and after drying receive the sample facies saliva without displacement of the center of crystallization and without damaging the texture of the picture facies saliva, where the ratio of the Central and peripheral zones visibile and corresponds to the state of the organism.

- the use of an insulating cover device of a portable laboratory during the drying drops mixed with saliva prevents the ingress of foreign particles into the drop, which avoids artifacts.

- reducing the number of false results if the instrument is portable laboratory reaches 50%. The results provided in table 1.

the method is technically available and requires no special skills and labor costs.

Example 1.

Child B., age 9, with a compensated form of caries, from the category of healthy children, with the index of the CPU 1, Hygienic index on green-Vermilion 0.2 points, with good oral hygiene. Using sterile swabs was extracted oral fluid in the amount of 2.0 ml and then receiving the receiving facies saliva from 0,02 ml drops mixed with saliva, in accordance with the proposed method applied on a glass slide and drying with the use of portable laboratory device. Received standard quality sample facies saliva height 1.0 mm, diameter 4.0 mm, form smooth rounded without distortion, and the standard sample facies saliva without displacement of the center of crystallization and without damaging the texture of the picture facies. For the Central zone is characterized by a clear illustration of the formation of large elongated kristallograficheskih structures coming from the center drops, fused among themselves and with the tree and pteridophyte form. The ratio of the following areas: the Central area is 70%, the peripheral zone is 30%. The obtained data indicate the absence of General pathology and pathology of the oral cavity that corresponds to the data records.

Example 2.

Child Of N., 12 years with decompensated form of caries, from the category of frequently ill children, the index of the CPU 9, hygienic index on the green-Vermilion 2.3 points with poor oral hygiene. Using sterile swabs was extracted mixed with saliva in the amount of 2.0 ml, and then the subsequent figures facies saliva from 0,02 ml drops mixed with saliva on the method proposed by: C. N. Shabalina, S. N. Athinai, 2001, without using the device as a portable laboratory. In R. the result obtained nonstandard saliva sample height 1.0 mm, diameter of 10.0 mm, the shape of the drop is deformed edge, distorted, and after drying in an incubator received a non-standard sample facies saliva with offset center of crystallization and a violation of the texture picture facies, which made it impossible to identify the important parameter in the diagnosis is the ratio of the Central and peripheral zones, which contributed to obtaining false results.

Example 3.

Child Of N., 12 years with decompensated form of caries, from the category of frequently ill children, the index of the CPU 9, hygienic index on the green-Vermilion 2.3 points with poor oral hygiene. Using sterile swabs was extracted oral fluid in the amount of 2.0 ml, and then the subsequent figures facies saliva from 0,02 ml drops of mixed saliva in accordance with the proposed method. At the beginning of the received standard quality saliva sample height 1.0 mm, diameter 5.0 mm, smooth round shape, and after drying using the device received the standard sample facies saliva without displacement of the center of crystallization and without damaging the texture of the picture facies. The Central zone consists of amorphous pattern over the entire area drops reviewed a large number isometrically located crystalline structures stellate and irregular shape. Identifies the self destruction of the crystals of the picture. The ratio of areas in percents: Central area 90%, the peripheral area is 10%. Indicating that somatic disorders in a patient.

Table 1
The method of obtaining standard quality sample facies saliva for crystallography
Method of crystallographyThe number of studied facies saliva (abs. numbers)The number of unsatisfactory results (abs. numbers)The number of unsatisfactory results (in percentages)The probability P
Crystallography saliva method prototype (C. N. Shabalin, S. N. Shatokhina, , Moscow, 2001)30019063%±m<0,05
Crystallography of saliva by the proposed method with the use of portable devices laboratory "Crystallina"3005016,6%±m<0,05

The method of obtaining standard quality sample facies of saliva in a crystal is ographie, includes a sampling of mixed saliva in a sterile tube with a volume of 2.0 ml, followed by the application of drops mixed with saliva on a glass slide and drying, characterized in that the mixed saliva pre-centrifuged for 20 minutes at 3000 rpm, it takes a set amount of 0.02 ml was centrifuged saliva using a micropipette and applied on a glass slide, placed on the surface of the portable device laboratory, established at levels X and Y, relative to the horizon, with the receipt of standard shape and size drops mixed with saliva height of 1.0 mm, a diameter of from 4.0 to 5.0 mm, round shape, in a further drop also dried on the surface of the portable device laboratory at room temperature +18...+25°C under an insulating lid for 5 hours to obtain figures facies saliva without distortion of the crystal pattern and without displacement of the center of crystallization.



 

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