Method for selecting donors of blood plasma applicable for treating patients with infectious diseases and postoperative, injury and burn complications

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to transfusion medicine, and is applicable in selecting blood plasma donors. That is ensured by sampling blood plasma to be analysed for antimicrobial peptides defensins. If the male defensin titre is from 180 to 545 ng/ml, and the female one is from 128 to 386 ng/ml, a donor is considered to be suitable for blood plasma drawing.

EFFECT: invention enables using the donor plasma in treating the patients suffering from infectious diseases and postoperative, injury and burn complications.

 

The invention relates to medicine, in particular to hemotransfusion, and can be used in the selection of donors, to obtain blood plasma, is effective in the treatment of patients with infectious diseases and complications after surgery, trauma and burns.

The known method of selection of donors by determining bacterial antibodies in their blood plasma, a widely used laboratory offices stations (offices) of blood transfusion, in accordance with the plan of acquisition and the anticipated therapeutic interventions in institutions following infection with significant therapeutic antibody titre:

- antistafilokokkovi in titer of 1:1280 and above;

- anticlimatically in titer of 1:160 and above;

- antisinegnoynym in titer of 1:160 and above;

- antiproteinase in titer of 1:160 and above;

- anticlericalism in titer of 1:160 and above (A. V. Chechetkin et al., 2008 / HOWTO "Receipt and clinical use of immune plasma in military medical facilities").

In practice, the production of transfusion was used two solutions to the issue of ensuring immune plasma. This is a targeted immunization of donors and screening of antibacterial antibodies in donors, allowing you to select the samples of fresh frozen plasma with antibodies for therapeutic titers (titers above).

The main pokazatelstva plasma samples of donors is the value or a meaningful therapeutic antibacterial antibodies titer (AT) for the provision of Transfusiology care. The percentage of detection AT the human donors ranged from 12 to 73% and depended on sex, age, time of year and seasonal work. The control activity of plasma donors with a standard immune serum was performed 1 time per 3-4 weeks. Plasma samples subsequently subjected to storage. If you are freezing for 6 hours after donating blood, the plasma is considered frozen (A. V. Chechetkin et al., 2007 / Method. the instructions - "Organization of karantinizatsii fresh frozen plasma in military medical facilities"), and may also be lyophilized in a timely manner and stored in a dry condition. Its application is important in the military field, but also in remote areas away from major medical institutions.

Since 2010, due to the lack of testing on the previously mentioned us infection prompted us to seek a new approach in donor screening to determine natural antibacterial antibodies in the plasma, it is necessary to provide medical assistance and targeted use of donor plasma a certain category of patients.

Finding a highly effective means of treatment and prevention of various infections to the blood service has now become particularly relevant and urgent task of practical health care.

The use palimony plasma advisable �ri verified nosocomial infections (Pseudomonas aeruginosa, Staphylococcus, Streptococcus, etc., pyogenic flora), burn disease, complicated by acceding secondary purulent infection, and in the clinic of internal diseases (severe bacterial pneumonia, septicemia, etc.).

The disadvantage of this method are the duration and the difficulty of identifying donors, and the lack of a legal framework practical implementation in the clinic to the above methods of treatment.

The aim of the invention is to improve the efficiency of treatment of patients with infectious diseases and complications after surgery, trauma and burns due to the use of donor blood plasma containing antimicrobial peptides (defensin), which have microbicides, chemotactic, immunomodulatory and cytotoxic activity without deliberate immunization of donors.

The objective is achieved by examining immunological parameters of blood, and reveal the content of antimicrobial peptides defensins in plasma and blood sampling was carried out in persons in the plasma which they are held at a titer of from 180 to 545 ng/ml in men, from 128 to 386 ng/ml in females.

The method is implemented as follows.

Conduct research on immunological parameters of blood donors, identify the content of antimicrobial peptides defensins and blood sampling for use with l�treatment of seriously ill patients is carried out in persons in the plasma which they are held at a titer of from 180 to 545 ng/ml in men, from 128 to 386 ng/ml in females.

Practical example 1. Donor - I. (male) titer of antimicrobial peptides amounted 550,6 ng/ml; blood plasma is directed to the clinic of military surgery for the treatment of seriously wounded, delivered from Dagestan

Practical example 2. Donor - K. (male) titer of antimicrobial peptides amounted to 33.1 ng/ml; blood plasma is directed to the clinic of faculty therapy.

Practical example 3. Donor - L. (woman) titer of antimicrobial peptides was at 228.9 ng/ml; blood plasma is directed to the clinic of military-field therapy for the treatment of a patient with severe poisoning.

Practical example 4. Donor - I. (woman) titer of antimicrobial peptides amounted 116,1 ng/ml; blood plasma is directed to the clinic of obstetrics and gynecology.

At present, antimicrobial peptides represent the focus of attention for a wide range of experts, including considering such connections, as an alternative to antibiotic drugs. To resist pathogenic microorganisms, scientists are developing new types of antibiotics, which are essentially derivatives of the old ones. Antimicrobial peptides are inferior to antibiotics in terms of efficacy, but are faster, destroying bacteria that are immune to antibiotics (Garsia A,E, Osaau G, Tran PA, Yuan J, Selsted ME, Infect Immun. 2008 Dec. 76 (12):5883-91).

Peptides are continuously formed even in a stable state of the body. A. E. Abaturov. The journal "child Health", 3 (38) 2012 indicates that even at low concentrations of defensins inhibit adenovirus infection.

Antimicrobial peptides-defensin - multifunctional cationic peptides of human rights and are short molecules with a length of 12 to 50 amino acids have an average molecular weight of 4 KD, perform complex functions in the maintenance and regulation of the immune response to viral and other microbial antigenic aggression through direct effects on the cell membrane of the microorganism that causes the destruction of cells and also participate in the regulation of the process of antigen presentation.

Medicines on the basis of antimicrobial antibodies has not yet been established. At the same time the content of defensins in plasma (a protein of human neutrophils) is interpreted as an indicator of the presence of high immunoprotective functions.

B. V. Pinegin, A. S. Budikhina in the journal "Immunology" 2008, No. 5, pp. 317-320 indicate that antimicrobial peptides (defensin) are components of the immune system and provide a first line of defense of an organism against various pathogens.

1. The advantages of the method:

- research time 3.5 hours;

- minimum measured concentration�tion 156 PG/ml;

- the measured range 156-10000 PG/ml;

- sample volume of 100 μl per well;

- the availability of test systems for research.

2. The principle of the method.

Based on the "sandwich" method, enzyme-linked immunosorbent assay with the use of a diagnostic kit HNP 1-3 (defensive neutrophils) by ELISA.

2. Preparation of samples of blood donors to the research demands strict implementation of all recommendations and methodical instructions.

We examined 88 human donors aged 21-34 years, of which 81 were male, 7 were female.

Obtained samples of blood donors were stored at +2-4°C. the plasma Separation was performed by centrifugation no later than 20 minutes after exposee blood at 1500 g at 4°C for 15 min and the Plasma was collected into clean polypropylene tubes, carefully, without touching the cell pellet. Re-centrifuged at the same conditions. The most reliable results were obtained when using as anticoagulant heparin or EDTA.

Samples before the study was stored at -70°C in polypropylene tubes. Storage at -20°C adversely affects the extraction defensins HNP 1-3.

Examination of samples of donated blood was performed 24 hours after thawing and prior dilution of the supplied buffer and blending in polypropylene tubes.

W�ka reagents conducting study procedures were performed in accordance with the instruction.

Out of the 81 surveyed men have identified 20 defensin HNP 1-3 from 180 to 545 ng/ml - 24,7%, 2 women - from 128 to 386 ng/ml - 28,5%.

Method of selection of donors for plasma blood by immunological studies of blood, characterized in that in the plasma reveal the content of antimicrobial peptides defensins and at a titer of defensins from 180 to 545 ng/ml in males and 128 to 386 ng/ml in females perform blood collection.



 

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FIELD: medicine.

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28 cl, 5 ex, 23 dwg, 5 tbl

FIELD: medicine.

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1 ex

FIELD: medicine.

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1 tbl, 3 ex

FIELD: medicine.

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3 tbl, 3 ex

FIELD: medicine, biology.

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3 ex

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1 ex, 1 tbl

FIELD: medicine.

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EFFECT: higher accuracy of detection.

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EFFECT: higher efficiency of detection.

2 ex

FIELD: medicine.

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EFFECT: enhanced accuracy of prediction.

FIELD: medicine, medicinal immunology.

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EFFECT: improved method for assay.

5 tbl, 1 ex

FIELD: medicine.

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EFFECT: high accuracy of diagnosis.

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