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.

SUBSTANCE: invention relates to medicine, namely to pediatrics, and can be applied for predicting reduction of child's health level at the age of 12-16. For this purpose an hour after meal and 15 minutes before mouth fluid sampling oral cavity is washed with 0.9% sodium chloride solution. Analysis of child's mouth fluid with determination of ratio of immunoglobulin A concentration to immunoglobulin G concentration is performed. Analysis is carried out twice with difference in 14 days. In case of 1.4 fold and higher reduction of immunoglobulin A concentration to immunoglobulin G concentration ratio during second analysis child is allocated to group of risk of health level reduction.

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2 ex

FIELD: medicine.

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4 ex

FIELD: medicine.

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28 cl, 5 ex, 23 dwg, 5 tbl

FIELD: medicine.

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EFFECT: method enables assessing the functional body reserves with the use of combined characteristics including the adaptation body potential, immune reactivity and immune cell interaction.

1 ex

FIELD: medicine.

SUBSTANCE: DNA is recovered from peripheral venous blood. Genetic polymorphisms of tumour necrosis factor α (-308 G/A TNFα), tumour necrosis factor 1 receptor (+36 A/G TNFR1), interferon-inducible T-cell chemoattractant (A/G I-TAC), interleukin 1A (-889 C/T IL-1A), lymphotoxin α (+250 A/G Ltα) are typed by polymerase chain reaction. A high risk of developing hyperplastic processes of endometrium is predicted if detecting a combination of alleles -308 G TNFα, +36 A TNFR1, A I-TAC, -889 T IL-1A and/or a combination of alleles +36 A TNFR1, A I-TAC, -889 T IL-1A and/or a combination of alleles -308 G TNFα,+250 G Ltα, -889 T IL-1A.

EFFECT: higher accuracy.

2 dwg, 1 tbl, 1 ex

FIELD: medicine.

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1 tbl, 3 ex

FIELD: medicine.

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3 dwg, 2 ex

FIELD: medicine.

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2 dwg, 2 tbl, 1 ex

FIELD: medicine.

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1 tbl, 2 ex

FIELD: medicine.

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3 ex, 1 tbl, 5 dwg

FIELD: medicine, ophthalmology.

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EFFECT: higher accuracy of prediction.

2 ex

FIELD: medicine, medicinal microbiology.

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EFFECT: improved assay method.

3 tbl, 3 ex

FIELD: medicine, biology.

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3 ex

FIELD: medicine, cardiology.

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EFFECT: higher accuracy of prediction.

FIELD: medicine, parasitology.

SUBSTANCE: one should carry out immunoenzymatic assay to detect diagnostic optic density and that of labeled immune complex in a plot's hole with tested serum measured in conventional units at wave length being 492 nm. One should calculate coefficient of antibodies concentration measured in conventional units by the following formula: CAC = (Odtsh - Odd) x 100, where CAC - coefficient of antibodies concentration, Odtsh - optic density of the hole with tested serum, Odd - diagnostic value of optic density, 100 - coefficient of serumal dilution. By CAC value one should detect the titer of antibodies to Lamblia intestinalis antigens to interpret results of the trial. The method enables to study the dynamics of disease flow.

EFFECT: higher efficiency and accuracy of diagnostics.

1 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: the present innovation deals with studying and treating diseases of inflammatory, autoimmune and degenerative genesis. One should perform sampling of heparinized blood followed by its sedimentation to obtain blood plasma with leukocytes and centrifuging to isolate the latter which are washed against erythrocytic and serumal admixtures, and, also, it deals with calculating the number of cells in samples out of leukocytic suspension after incubation (B) for 1.5 h at 37 C in holes of plastic microplotting board, out of leukocytic suspension one should additionally prepare two samples, one should be applied to calculate total number of leukocytes before incubation (A), the second sample undergoes incubation at the same mode at addition of autoserum to calculate the number of cells remained after incubation (C). One should state upon adhesive properties of leukocytes by the index of spontaneous adhesion (D), where D=(A-B)/B.100%, and effect for enhanced cellular adhesion under the impact of autoserum should be detected by the value of K=(B-C)/C.100% at K ≥ 30%, where B - C - the number of cells undergone additional adhesion after addition of autoserum. The present innovation widens functional possibilities of the suggested method due to obtaining additional values depicting adhesive properties of blood leukocytes.

EFFECT: higher accuracy of detection.

FIELD: medicine, immunology.

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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.

SUBSTANCE: method involves determination of heterophilic antibodies in human serum blood by the Paul-Bunnel's method relatively the level of circulating immune complexes, complement-activating properties of heterophilic antibodies by incubation of standardized ram erythrocytes with 0.8% serum for 30 ± 5 min and the following measurement of the erythrocytes lysis degree. The measurement of the effector function coefficient of heterophilic antibodies is carried out by the complement system Keff.f.h.a.-c.s. by the formula: Keff.f.h.a.-c.s. = Y/Tg.a. wherein Y means a lysis degree, %; Tg.a. means a reverse titer of heterophilic antibodies to ram erythrocytes. The damage assay is carried out by comparison of the immune status with the relative level of circulating immune complexes in serum. Method provides detection of preclinic from of immunodeficiency and autoimmune diseases that opens the possibility for their prophylaxis at most early stages of development. Invention can be used for assay of damage in the immune status in human serum blood.

EFFECT: improved method for assay.

5 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method involves concurrently examining anti-inflammatory IL-4 level in blood serum and lacrimal fluid. The value being within the limits of 60-70 pg/l in blood serum and 5-15 pg/l in lacrimal fluid, disease prognosis is considered to be unfavorable. The IL-4 concentration being within the limits of 90-100 pg/l in blood serum and 20-30 pg/l in lacrimal fluid, disease prognosis is considered to be favorable.

EFFECT: high accuracy of diagnosis.