CN112023071A - A method for inactivating pathogenic microorganisms in blood products - Google Patents

Abstract

The invention belongs to the technical field of blood product pathogen inactivation, and discloses a method for inactivating pathogenic microorganisms in a blood product, which comprises the following steps: pre-treating the blood product before irradiation; irradiating the pretreated blood product with X-rays and gamma-rays; blood products were infused after irradiation. According to the method, the blood product is irradiated by combining X rays and gamma rays, so that pathogenic microorganisms containing DNA such as bacteria in the blood product can be inactivated, and transfusion transmission infection is avoided.

Description

A method for inactivating pathogenic microorganisms in blood products

technical field

The present disclosure belongs to the technical field of pathogen inactivation in blood products, and in particular relates to a method for inactivating pathogenic microorganisms in blood products.

Background technique

Blood transfusion therapy is an important and irreplaceable clinical treatment method. Transfusion of blood products not only saves the patient's life, but also has many adverse blood transfusion reactions. Among them, transfusion-transmitted infection is one of the serious adverse blood transfusion reactions, which often causes irreversible consequences to the recipient. Although WHO has put forward a blood safety strategy and a series of recommendations, including questionnaires before blood donation, detection of some pathogens, and suspension of blood collection in the event of a local outbreak of infectious diseases. However, due to the long time range and subjectivity of the questionnaire before blood donation, some asymptomatic infected persons cannot be found through questionnaire screening; due to the detection range, sensitivity and window period of blood products pathogenic microorganisms, some of them carry pathogenic microorganisms. The blood cannot be detected, which affects the safety of blood transfusion.

Therefore, pathogenic microorganism infection caused by blood transfusion has been accompanied by blood transfusion treatment worldwide and has become an important blood transfusion adverse event.

SUMMARY OF THE INVENTION

In view of the deficiencies of the prior art, the purpose of the present disclosure is to provide a method for inactivating pathogenic microorganisms in blood products, which solves the problems mentioned in the background art.

The purpose of the present disclosure can be achieved through the following technical solutions:

A method for inactivating pathogenic microorganisms in blood products, comprising the steps of:

Step 1. Pre-treatment of blood products before irradiation;

Step 2, irradiating the pretreated blood products with X-rays and γ-rays in combination;

Step 3: All blood products are infused after irradiation.

Further, the total irradiation dose of the X-rays and the γ-rays is 25Gy˜50Gy.

Further, the X-ray irradiation dose is 25Gy, and the γ-ray irradiation dose is 25Gy.

Further, the X-ray irradiation dose is 15 Gy, and the γ-ray irradiation dose is 35 Gy.

Further, the X-ray irradiation dose is 35 Gy, and the γ-ray irradiation dose is 15 Gy.

Further, the X-ray irradiation dose is 12.5Gy, and the γ-ray irradiation dose is 12.5Gy.

Beneficial effects of the present disclosure:

The present disclosure can inactivate DNA-containing pathogenic microorganisms such as bacteria in blood products by irradiating blood products in combination with X-rays and γ-rays, thereby avoiding transfusion-transmitted infections.

Detailed ways

The following will clearly and completely describe the technical solutions in the embodiments of the present disclosure. Obviously, the described embodiments are only a part of the embodiments of the present disclosure, rather than all the embodiments. Based on the embodiments in the present disclosure, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present disclosure.

A method for inactivating pathogenic microorganisms in blood products, comprising the steps of:

Step 1. Pretreatment of blood products before irradiation

In general, blood products are stored at normal storage temperature, red blood cells are stored at 2°C to 6°C, platelets are stored at 20°C to 24°C, and cryoprecipitate and plasma are stored below -20°C;

Thaw plasma and cryoprecipitate products at 37°C;

Carry out a cross-matching test on red blood cell products, and perform a cross-matching test on platelets. If the cross-matching blood matches, then blood can be transfused. Select the blood products that match the blood and wait for the next step of irradiation treatment;

Step 2. X-ray and γ-ray irradiation of blood products

Irradiate pretreated blood products with a combination of X-rays and gamma rays;

There is a certain difference in the dose rate and irradiation time of different instruments. The irradiation time and dose rate are determined according to the actual situation, and the final total irradiation dose can reach 25Gy~50Gy;

X-rays and γ-rays are rays with strong penetrating ability, and the ionizing radiation generated by them can cause permanent damage to cellular DNA, resulting in changes in molecular structure and loss of biological activity, which can inactivate pathogenic microorganisms;

According to the characteristics of X-rays and γ-rays, and taking into account the differences in wavelength, frequency, and penetrating power, two kinds of rays are used in combination to irradiate blood products. The inactivation effect of pathogenic microorganisms;

At the same time, since the active components in blood products such as red blood cells, platelets, fibrinogen, and coagulation factors do not contain DNA, their sensitivity to ionizing radiation generated by X-rays and γ-rays is much lower than that of pathogenic microorganisms containing DNA. Low-dose irradiation has less effect on the active components of blood products. Therefore, by irradiating a suitable dose of radiation, the pathogenic microorganisms in blood products can be inactivated to a large extent, and the effective components of the blood can be hardly affected, which can ensure the safety and effectiveness of blood transfusion to a large extent.

The present disclosure can inactivate DNA-containing pathogenic microorganisms such as bacteria in blood products by irradiating blood products together with X-rays and γ-rays, avoid transfusion-transmitted infections, and solve the problem that blood products such as red blood cells, platelets, and cryoprecipitates cannot be inactivated. The technical blank of live pathogenic microorganisms is of great significance to ensure the safety of blood transfusion; at the same time, the functions of red blood cells, platelets, fibrinogen, coagulation factors and other active ingredients in blood products are basically unchanged, ensuring the effectiveness of blood transfusion.

Step 3. Blood products are infused after irradiation

The method for inactivating pathogens in blood products of the present disclosure has simple steps, short time required, and high work efficiency, and no other substances need to be added to the blood products, thereby avoiding artificial pollution and residual toxic and harmful substances.

Example 1

A method for inactivating pathogenic microorganisms in blood products, comprising the steps of:

Step 1. Pretreatment of blood products before irradiation

In general, blood products are stored at normal storage temperature, red blood cells are stored at 2°C to 6°C, platelets are stored at 20°C to 24°C, and cryoprecipitate and plasma are stored below -20°C;

Thaw plasma and cryoprecipitate products at 37°C;

Carry out a cross-matching test on red blood cell products, and perform a cross-matching test on platelets. If the cross-matching blood matches, then blood can be transfused. Select the blood products that match the blood and wait for the next step of irradiation treatment;

Step 2. X-ray and gamma irradiation of blood products

The pretreated blood products are irradiated under X-rays and γ-rays, and the X-ray irradiation dose is 25Gy, and the γ-ray irradiation dose is 25Gy;

Step 3. Infuse the irradiated blood product.

Embodiment 2

A method for inactivating pathogenic microorganisms in blood products, comprising the steps of:

Step 1. Pretreatment of blood products before irradiation

In general, blood products are stored at normal storage temperature, red blood cells are stored at 2°C to 6°C, platelets are stored at 20°C to 24°C, and cryoprecipitate and plasma are stored below -20°C;

Thaw plasma and cryoprecipitate products at 37°C;

Carry out a cross-matching test on red blood cell products, and perform a cross-matching test on platelets. If the cross-matching blood matches, then blood can be transfused. Select the blood products that match the blood and wait for the next step of irradiation treatment;

Step 2. X-ray and gamma irradiation of blood products

The pretreated blood products are irradiated under X-rays and γ-rays, the X-ray irradiation dose is 35Gy, and the γ-ray irradiation dose is 15Gy;

Step 3. Infuse the irradiated blood product.

Embodiment 3

A method for inactivating pathogenic microorganisms in blood products, comprising the steps of:

Step 1. Pretreatment of blood products before irradiation

In general, blood products are stored at normal storage temperature, red blood cells are stored at 2°C to 6°C, platelets are stored at 20°C to 24°C, and cryoprecipitate and plasma are stored below -20°C;

Thaw plasma and cryoprecipitate products at 37°C;

Carry out a cross-matching test on red blood cell products, and perform a cross-matching test on platelets. If the cross-matching blood matches, then blood can be transfused. Select the blood products that match the blood and wait for the next step of irradiation treatment;

Step 2. X-ray and gamma irradiation of blood products

The pretreated blood products are irradiated under X-rays and γ-rays, the X-ray irradiation dose is 15Gy, and the γ-ray irradiation dose is 35Gy;

Step 3. Infuse the irradiated blood product.

Embodiment 4

A method for inactivating pathogenic microorganisms in blood products, comprising the steps of:

Step 1. Pretreatment of blood products before irradiation

In general, blood products are stored at normal storage temperature, red blood cells are stored at 2°C to 6°C, platelets are stored at 20°C to 24°C, and cryoprecipitate and plasma are stored below -20°C;

Thaw plasma and cryoprecipitate products at 37°C;

Carry out a cross-matching test on red blood cell products, and perform a cross-matching test on platelets. If the cross-matching blood matches, then blood can be transfused. Select the blood products that match the blood and wait for the next step of irradiation treatment;

Step 2. X-ray and gamma irradiation of blood products

The pretreated blood products are irradiated under X-rays and γ-rays, the X-ray irradiation dose is 12.5Gy, and the γ-ray irradiation dose is 12.5Gy;

Step 3. Infuse the irradiated blood product.

Test Example 1

In this test example, the bacterial inactivation effect in plasma was verified;

Add 9 ml of plasma to the Staphylococcus epidermidis suspension, mix well, and add 9 sterilized tubes of 1 ml each. Plasma containing bacterial liquid was randomly divided into 9 groups, including control group, X15γ35, X25γ25, X35γ15, γ50, X50, X25, X12.5γ12.5, γ25, and the irradiation dose was the same as the group name;

The X-ray dose rate is 1.246Gy/min, and the irradiation time of X15γ35, X25γ25, X35γ15, X50, X25, X12.5γ12.5 groups is 722 seconds, 1203 seconds, 1685 seconds, 2408 seconds, 1203 seconds, and 602 seconds, respectively. The radiation doses of rays are 15Gy, 25Gy, 35Gy, 50Gy, 25Gy, 12.5Gy;

The γ-ray dose rate is 4Gy/min, and the irradiation time of X15γ35, X25γ25, X35γ15, γ50, X12.5γ12.5, and γ25 groups is 525 seconds, 375 seconds, 225 seconds, 750 seconds, 187 seconds, and 375 seconds, respectively. The radiation doses are 35Gy, 25Gy, 15Gy, 50Gy, 12.5Gy, 25Gy;

The control group and the irradiation group were subjected to bacterial counts immediately after irradiation, 24 hours, 48 hours, and 72 hours, respectively, and the specimens to be tested were stored at 4 °C. The test results are shown in Table 1;

The results showed that the bacterial count in the irradiation group was less than that in the control group, indicating that radiation irradiation had the effect of killing pathogenic microorganisms;

The irradiation dose of X12.5γ12.5 group has the effect of killing pathogenic microorganisms; the sterilization effect of X15γ35, X25γ25 and X35γ15 is better than that of X12.5γ12.5X, that is, the sterilization effect is enhanced with the increase of the irradiation dose;

The bacterial count in the combined X-ray and γ-ray group (x25γ25, x12.5γ12.5) was less than that in the same dose of X-ray and γ-ray group alone. Combined X-ray and γ-ray irradiation of plasma has better inactivation effect on bacteria in plasma;

Studies have shown that the number of initially contaminated bacteria in blood is relatively small, rarely exceeding 10 CFU/ml, and the number of bacteria added in this experiment far exceeds the number of possible contamination. The effect will be better.

Table 1: Plasma bacterial counts before and after irradiation

Test Example 2

In this test case, the function test of plasma after irradiation;

Divide 20 ml of plasma into 4 tubes, each with a capacity of 5 ml, and randomly assign them to 4 groups, including control group, X25γ25, X50, and γ50, and the irradiation dose is the same as the group name;

The X-ray dose rate was 1.267 Gy/min, and the γ-ray dose rate was 1.704 Gy/min. The irradiation time of the control group, X25γ25, X50, and γ50 groups was X-ray 1185 seconds combined with γ-ray 880 seconds, 2370 seconds, and 1760 seconds, respectively, and the irradiation dose was X-ray 25Gy combined with γ-ray 25Gy, X-ray 50Gy, and γ-ray 50Gy. ;

The control group and the irradiation group were tested for blood coagulation function and thrombelastography immediately and 24 hours after irradiation, respectively, and the plasma to be tested was stored at 4°C. The test results are shown in Table 2;

The results showed that there was no significant difference in plasma coagulation function between the control group and the irradiation group, and there was no significant difference in plasma coagulation function before and after irradiation in the same dose of single radiation and combined radiation groups, and low dose radiation had no significant effect on coagulation factors.

Table 2: Plasma coagulation function and thromboelastography before and after irradiation

Test Example 3

In this test example, the hemolysis-related indexes before and after the irradiation of red blood cells were detected;

Add 36 ml of red blood cell products that have been stored for about 10 days into 9 sterilized tubes on average, 4 ml per tube. Randomly assigned to 9 groups, including control group, X15γ35, X25γ25, X35γ15, γ50, X50, X25, X12.5γ12.5, γ25, and the irradiation dose has the same group name;

The X-ray dose rate is 1.246Gy/min, and the irradiation time of X15γ35, X25γ25, X35γ15, X50, X25, X12.5γ12.5 groups is 722 seconds, 1203 seconds, 1685 seconds, 2408 seconds, 1203 seconds, and 602 seconds, respectively. The radiation doses of rays are 15Gy, 25Gy, 35Gy, 50Gy, 25Gy, 12.5Gy;

The γ-ray dose rate is 4Gy/min, and the irradiation time of X15γ35, X25γ25, X35γ15, γ50, X12.5γ12.5, and γ25 groups is 525 seconds, 375 seconds, 225 seconds, 750 seconds, 187 seconds, and 375 seconds, respectively. The radiation doses are 35Gy, 25Gy, 15Gy, 50Gy, 12.5Gy, 25Gy;

The control group and the irradiation group were tested for plasma potassium, bilirubin, aspartate aminotransferase, lactate dehydrogenase, creatine kinase, glucose and free hemoglobin immediately after irradiation, 24 hours, 48 hours, and 72 hours, respectively. For testing, the samples to be tested were stored at 4°C, and the test results were shown in Table 3;

The results showed that there was no significant change in bilirubin, aspartate aminotransferase, lactate dehydrogenase, creatine kinase, glucose and free hemoglobin before and after irradiation; there was no significant change in K ions in each group immediately after irradiation; Potassium ions in the 24-hour, 48-hour and 72-hour irradiation groups increased; at 48 hours and 72 hours after irradiation, the increase in K ions in the γ25 and γ50 groups was correspondingly higher than that in the X25 and X50 groups; X15γ35X, X25γ25, X35γ15 groups The increase range of K ion was basically between the γ50 and X50 groups, and the increase range of the X12.5γ12.5 group was between the γ25 and X25 groups; the difference in potassium ions between the irradiation group and the control group increased with the prolongation of red blood cell storage time.

Table 3: Detection of erythrocyte hemolysis-related indicators before and after irradiation

In the description of this specification, description with reference to the terms "one embodiment," "example," "specific example," etc. means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one aspect of the present disclosure. in one embodiment or example. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

The foregoing has shown and described the basic principles, key features, and advantages of the present disclosure. Those skilled in the art should understand that the present disclosure is not limited by the above-mentioned embodiments. The above-mentioned embodiments and descriptions only illustrate the principles of the present disclosure. Without departing from the spirit and scope of the present disclosure, the present disclosure will also include Various changes and modifications are intended to fall within the scope of the claimed disclosure.

Claims (6)

1. A method of inactivating a pathogenic microorganism in a blood product, comprising the steps of:

step one, performing pretreatment before irradiation on a blood product;

step two, irradiating the pretreated blood product by jointly using X rays and gamma rays;

and step three, infusing the blood products after irradiation.

2. The method of claim 1, wherein the total dose of X-rays and gamma-rays is from 25Gy to 50 Gy.

3. The method of claim 1, wherein the X-ray radiation dose is 25Gy and the gamma-ray radiation dose is 25 Gy.

4. The method of claim 1, wherein the X-ray radiation dose is 15Gy and the gamma radiation dose is 35 Gy.

5. The method of claim 1, wherein the X-ray radiation dose is 35Gy and the gamma-ray radiation dose is 15 Gy.

6. The method of claim 1, wherein the X-ray radiation dose is 12.5Gy and the gamma-ray radiation dose is 12.5 Gy.