Method for keeping viable blood cells for purposes of chemoluminescent analysis

FIELD: medicine.

SUBSTANCE: method for keeping viable blood cells for purposes of chemoluminescence analysis involves blood sampling and preserving with the blood sampled in disposable plastic test tubes and Faglucide® used as a haemopreserving agent in the ratio 1:6; the prepared blood is kept at temperature 2 to 6°C.

EFFECT: prolonged period of keeping viable leukocytes for purposes of chemoluminescence analysis, maintained the morphofunctional integrity and the rheological blood properties in cattle for 168 hours and the pH value at a physiological level that enables blood transportation for diagnostic researches from distant regions to laboratories.

2 tbl

 

The invention relates to biology, namely, immunology, and can be used for chemiluminescent analysis in veterinary medicine.

For preservation of blood is applied to False®, which is used in medicine only for the preservation of donor blood, but not used in veterinary medicine. False® solution geoconservation issued of JSC "Joint-Kurgan society of medical drugs and products "Synthesis". In 1 liter of solution contains: adenine 0.34 g; dextrose (calculated on anhydrous) 30.0 g; citric acid monohydrate 1,365 g; sodium orthophosphate dodecahydrate 7.5 g; sodium citrate (disubstituted) 14.0 g; water for injection up to 1 L.

As described in the literature closest to the invention is patent RU 2280689 C2, 05.05.2004 "Method of preservation of viable leukocytes". The method is as follows. Pre make the blood from 2.7% solution of Trilon B (solution of sodium salt of ethylenediaminetetraacetic acid) in tubes treated with 3% solution polymethylsiloxane fluid in chloroform. The destruction of red blood cells is carried out by hypotonic lysis. Centrifuged at 1200 rpm for 15 minutes Additional leukocytes washed with 0.02% Trilon B. Centrifuged under the above mode, the supernatant liquid is poured. Sediment suspension cells resuspended in abilitybased the solution of the following composition; concentrate Hanks without phenol red in the ratio with water 1:9.

However, this method has the following disadvantages: the ratio of volume of 2.7% solution of trylon B and blood of cattle 1:10 is required to research the concentration of viable cells (90-95%) within four hours of taking blood samples (see table 1); Trilon B lowers the calcium level in the blood sample, which leads to a distortion of results.

The aim of the invention is the need to preserve the constancy of the parameters of the red blood cells of cattle and to increase the retention of viable cells (90-95%) for conducting a chemiluminescent analysis.

This objective is achieved in that for preserving blood is geoconsultants False®, which ensures the preservation of the functional usefulness of the blood of cattle within 7 days by maintaining physiological levels of ATP in erythrocytes, blood rheological properties and pH on the physiological level. Geoconsultants False® is mixed with blood in a ratio of 1:6.

The method is as follows. Produce blood in a disposable plastic tubes with geoconservation Perlucidum® in a ratio of 1:6, which are then placed in an insulated pin is iner with headelement and delivered to the laboratory. Harvested blood is stored at a temperature of from 2 to 6°C.

Using this method allows you to transfer blood from remote regions for diagnostic studies in the laboratory. Using Fallside® as geoconservation in the ratio of 1:6 allows to increase the retention of viable cells in the concentration of 90-95% for 168 hours after blood collection (see table 2), which is 42 times greater than when using the trylon B.

The method of preserving a viable blood cells for chemiluminescent studies, including taking blood samples and preservation, characterized in that the blood sampling is carried out in disposable plastic tubes and as geoconservation use False®in the ratio of 1:6, harvested blood is stored at a temperature of from 2 to 6°C.



 

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