阅读说明：本技术 全自动rh系统血型检测仪检测方法 (Detection method of full-automatic RH system blood type detector ) 是由 张志飞 常宗瑜 向川 沙俊宏 于 2021-06-28 设计创作，主要内容包括：本发明适用于血型检测技术领域,提供了一种全自动RH系统血型检测仪检测方法,包括如下步骤：将血液样本(EDTA抗凝全血)上机；上机后对红细胞悬液进行稀释,得到稀释液；再将稀释液加入微孔板中,微孔板中已经事先加入抗体,抗原抗体震荡混匀后离心,使抗原抗体充分凝集,再次震荡,最后拍照判读。借此,本发明能够实现血液的全自动检测,无需人工操作,检测时间短,效率高,且能够对阳性和弱阳性的血型进行精确的判读,避免由于检测不出弱阳性血型而耽误患者输血,造成患者无法救治的现象。(The invention is suitable for the technical field of blood type detection, and provides a detection method of a full-automatic RH system blood type detector, which comprises the following steps: processing a blood sample (EDTA anticoagulated whole blood) on a machine; diluting the erythrocyte suspension after the machine is operated to obtain a diluent; and adding the diluent into a microporous plate, adding the antibody into the microporous plate in advance, shaking and uniformly mixing the antigen and the antibody, centrifuging to ensure that the antigen and the antibody are fully agglutinated, shaking again, and finally photographing and interpreting. Therefore, the blood detection device can realize full-automatic detection of blood, does not need manual operation, has short detection time and high efficiency, can accurately judge the blood types of positive and weak positive, and avoids the phenomenon that the blood transfusion of a patient is delayed due to the fact that the blood type of the weak positive cannot be detected, so that the patient cannot be cured.)

1. A full-automatic RH system blood type detector detection method is characterized by comprising the following steps:

step one, blood sample is put on machine

Placing the centrifuged blood test tubes in the step one on a sample plate of a blood type detector, and scanning the two-dimensional codes on the blood test tubes through a control system of the blood type detector to record the patient information on each blood test tube into a system;

step two, adding the antibody into the microporous plate

Then adding the RH typing antibody into the microporous plate respectively;

step three blood sample dilution

Dripping normal saline into the deep hole plate through a control system of the blood type detector; then transferring the erythrocyte suspension in the blood test tube, and adding the erythrocyte suspension into the deep hole plate to which the normal saline is dripped; diluting the erythrocyte suspension by using normal saline to obtain a diluent;

step four antigen antibody mixing

Adding the diluent into a microplate dropwise added with the antibodies, and mixing the diluent with the five antibodies respectively; controlling the oscillator to work through a control system of the blood type detector, so that the diluent and the antibody are uniformly mixed;

step five centrifugation

Placing the micro-porous plate vibrated in the step four into a centrifuge of a blood type detector, and centrifuging the fully mixed antigen-antibody;

step six oscillation dispersion

Shaking the micro-porous plate subjected to centrifugation in the fifth step again to disperse the weak positive agglutinates;

step seven photographing interpretation

And taking out the micro-porous plate vibrated in the step six, then taking a picture, and judging the blood type of the blood according to the agglutination condition of the blood in the picture.

2. The method for testing a full automatic RH system blood type detecting instrument according to claim 1, wherein the micro well plate is a 96 well plate.

3. The method as claimed in claim 1, wherein the volume of each well of the microplate is 100-.

4. The method for testing a full automatic RH system blood type detecting instrument according to claim 1, wherein the amount of the five antibodies added to the microplate is 10-50 ul.

5. The method for testing a full automatic RH system blood type detecting instrument according to claim 1, wherein the volume of the red blood cell suspension in the diluent is 2-4%.

6. The method for testing the fully automatic RH system blood type detecting instrument according to claim 1, wherein the amount of the diluent added to the micro well plate is 10-50 ul.

7. The method as claimed in claim 1, wherein the oscillation time of the oscillator is 25-35s, the amplitude is 1-3mm, and the frequency is 500-3000 RPM.

8. The method as claimed in claim 1, wherein the relative centrifugal force of the centrifuge is 500g and the time is 0.8-1.5 min.

9. The method for testing the fully automatic RH system blood type detecting instrument according to claim 2, wherein the bottom of each well of the micro-well plate is flat bottom, U-shaped bottom or UV-shaped bottom.

Technical Field

The invention relates to the technical field of blood type detection methods, in particular to a detection method of a full-automatic RH system blood type detector.

Background

Blood type is the classification of blood, usually red blood cells, based on the presence of some inheritable antigenic material on the red blood cell membrane. Among the human blood group systems, there are 36 blood group systems that have been found and recognized by the international blood transfusion association, including ABO blood group system, Rh blood group system, MNS blood group system, P blood group system, etc., of which ABO blood group system and Rh blood group system are the most commonly used.

At present, the blood type detection mainly depends on manual detection of workers, and the detection efficiency is low, the time is long, and the labor cost is high; the ABO blood group positive typing and the Rh blood group detection are based on the agglutination reaction principle of erythrocyte antigen antibodies, the detection result is judged and read through a negative reaction pattern and a positive reaction pattern, the positive reaction is that the erythrocyte agglutination occurs, otherwise, the negative reaction is that the erythrocyte agglutination does not occur, and the detection method mainly comprises the following steps: centrifuging blood, separating out red blood cells, adding an antibody into the red blood cells, shaking to fully mix the red blood cells and the antibody, waiting for agglutination of the red blood cells and the antibody, and judging the blood type according to the agglutination condition; although the method is more conventional and more common in application, the method can only detect the positive blood type, but can not detect the weak positive blood type, and if the patient is in a critical state, the blood type judgment deviation can directly influence the life of the patient; in addition, the existing method generally takes more than 20min in the process of detecting blood type, the time is long, and the working efficiency is low.

The patent with publication number CN110146711A discloses a full-automatic blood type testing device and a blood type testing method thereof, which also apply the above conventional method, the precision of the testing result is not high, and the testing time is long.

In view of the above, the prior art is obviously inconvenient and disadvantageous in practical use, and needs to be improved.

Disclosure of Invention

Aiming at the defects, the invention aims to provide a full-automatic RH system blood type detector detection method which can realize full-automatic detection of blood, does not need manual operation, has short detection time and high efficiency, can accurately judge the blood types of positive and weak positive, and avoids the phenomenon that the transfusion of a patient is delayed due to the fact that the blood type of the weak positive cannot be detected, so that the patient cannot be cured.

In order to achieve the aim, the invention provides a full-automatic RH system blood type detector detection method, which comprises the following steps:

step one, blood sample is put on machine

And (3) placing the centrifuged blood test tube in the step one on a sample plate of the blood type detector, and scanning the two-dimensional code on the blood test tube through a control system of the blood type detector to enable the patient information on each blood test tube to be input into the system.

Step two, adding the antibody into the microporous plate

The microplate is now placed on a shaker, and the RH typing antibodies are then added to the microplate separately.

Step three blood sample dilution

Dripping normal saline into the deep hole plate through a control system of the blood type detector; then transferring the erythrocyte suspension in the blood test tube, and adding the erythrocyte suspension into the deep hole plate to which the normal saline is dripped; diluting the erythrocyte suspension by using normal saline to obtain a diluent.

Step four antigen antibody mixing

Adding the diluent into a microplate dropwise added with the antibodies, and mixing the diluent with the five antibodies respectively; and controlling the oscillator to work through a control system of the blood type detector, so that the diluent and the antibody are uniformly mixed.

Step five centrifugation

And (4) putting the micro-porous plate vibrated in the step four into a centrifuge of a blood type detector, and centrifuging the fully mixed antigen-antibody.

Step six oscillation dispersion

And (4) oscillating the micro-porous plate subjected to centrifugation in the fifth step again to disperse the weak positive agglutinate blocks.

Step seven photographing interpretation

And taking out the micro-porous plate vibrated in the step six, then taking a picture, and judging the blood type of the blood according to the agglutination condition of the blood in the picture.

According to the detection method of the full-automatic RH system blood type detector, the microporous plate is a 96-pore plate.

According to the detection method of the full-automatic RH system blood type detector, the bottom of each hole in the micropore plate is flat bottom, U-shaped bottom or UV-shaped bottom.

According to the detection method of the full-automatic RH system blood type detector, the volume of each hole in the micropore plate is 100-.

According to the detection method of the full-automatic RH system blood type detector, the adding amount of the five antibodies in the micropore plate is 10-50ul respectively.

According to the detection method of the full-automatic RH system blood type detector, the volume of the erythrocyte suspension in the diluent is 2-4%.

According to the detection method of the full-automatic RH system blood type detector, the addition amount of the diluent in the micropore plate is 10-50 ul.

According to the detection method of the full-automatic RH system blood type detector, the oscillation time of the oscillator is 25-35s, the amplitude is 1-3mm, and the frequency is 500-3000 RPM.

According to the detection method of the full-automatic RH system blood type detector, in the centrifugation process, the relative centrifugal force of the centrifuge is 250-500g, and the centrifugation time is 0.8-1.5 min.

The invention aims to provide a detection method of a full-automatic RH system blood type detector, which comprises the steps of firstly carrying out vibration mixing on a mixture of antigen and antibody on the full-automatic blood type detector, improving the mixing uniformity of the antigen and antibody, carrying out centrifugal operation after the antigen and antibody are uniformly mixed, leading positive and weak positive samples to be agglutinated under the action of centrifugal force, then carrying out vibration, leading the agglutination of the weak positive samples to be vibrated and dispersed, leading the positive agglutination to be still gathered together, and further distinguishing the blood of the positive and the weak positive samples. In conclusion, the invention can realize full-automatic detection of blood, does not need manual operation, has short detection time and high efficiency, can accurately judge the blood types of positive and weak positive, and avoids the phenomenon that the transfusion of a patient is delayed due to the fact that the blood type of the weak positive cannot be detected, so that the patient cannot be cured.

Drawings

FIG. 1 is a schematic view of the detection process of the present invention;

FIG. 2 is a photograph interpretation of the present invention;

FIG. 3 is a schematic view of the blood testing apparatus of the present invention;

in the figure, 1-sample plate, 2-shaker, 3-microwell plate, 4-antibody, 5-deep well plate, 6-centrifuge, 7-centrifuge.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to the accompanying drawings. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

The invention provides a full-automatic RH system blood type detector detection method, which comprises the following steps:

step one, blood sample is put on machine

On the artifical sample board 1 of placing blood type detector with the blood test tube after above-mentioned centrifugation, push sample board 1 manual work in the blood type detector, the control system through the blood type detector scans the two-dimensional code on the blood test tube, makes the patient information entry system on every blood test tube.

Step two, adding the antibody into the microporous plate

Through the control system of the blood type detector, the micropore plate 3 is placed on the oscillator 2, and then the RH typing antibody 4 is respectively added into the micropore plate 3. The micropore plate 3 is a 96-pore plate, the volume of each pore in the micropore plate 3 is 100-. The adding amount of the five antibodies 4 in the microporous plate 3 is 10-50ul respectively; 18 wells were added for each antibody 4, i.e. five antibodies 4 occupied 90 wells in total. The RH typing antibody 4 includes five types of D, C, C, E, E.

Step three blood sample dilution

Dripping physiological saline into the deep hole plate 5 through a control system of the blood type detector; then transferring the red blood cell suspension in the blood test tube through a control system of the blood type detector, and adding the red blood cell suspension into the deep hole plate 5 which is dripped with the physiological saline; diluting the erythrocyte suspension by using normal saline to obtain a diluent.

In the diluent, the volume of the erythrocyte suspension accounts for 2-4%, for example, 500ul of normal saline is firstly added into the deep-hole plate 5, then 20ul of erythrocyte suspension is added, and the mixture is blown and beaten evenly.

Step four antigen antibody mixing

Adding the diluent into the microporous plate 3 in which the antibodies 4 are dripped through a control system of the blood type detector, and mixing the diluent with the five antibodies 4 respectively; the control system of the blood type detector controls the oscillator 2 to work, so that the diluent and the antibody 4 are uniformly mixed, and the agglutination of the erythrocyte antigen and the antibody 4 is promoted. The addition amount of the diluent in the microporous plate 3 is 10-50 ul. The oscillation time of the oscillator is 25-35s, the amplitude is 1-3mm, and the frequency is 500-3000 RPM.

Step five centrifugation

Through the control system of the blood type detector, the micro-porous plate 3 which is vibrated in the step four is put into a centrifuge 6 of the blood type detector, and the fully mixed antigen and antibody 4 is centrifuged. Through the centrifugal action, the agglutinated antigen-antibody 4 can be fully gathered, the agglutination effect can be conveniently judged, and the blood type can be judged. In the process of centrifugation, the relative centrifugal force of the centrifuge 6 is 250-500g, and the centrifugation time is 0.8-1.5 min.

Step six oscillation dispersion

And (4) oscillating the micro-porous plate subjected to centrifugation in the fifth step again to disperse the weak positive agglutinate blocks. Because positive and weak positive clumps are aggregated in the centrifugation process, the positive and weak positive clumps cannot be distinguished only by centrifugation; the invention can shake apart the weak positive agglutinate after the centrifugation and shake again, and the positive agglutinate is always gathered, therefore, the positive and the weak positive agglutinate can be distinguished by the action of shaking again.

Step seven interpretation

And taking out the micro-porous plate 3 vibrated in the step six, then placing the micro-porous plate in a centrifuge 7 for photographing, and judging the blood type of the blood according to the agglutination condition of the blood in the photograph. According to the invention, the agglutination strength is judged according to the size of the erythrocyte clot in the micropores, and the agglutinated block which is gathered together can be dispersed by the action of concussion for the agglutination which shows weak positive. Therefore, the operation of centrifuging and then shaking can lead the method to detect the positive and weak positive blood types and the detection result to be accurate. When a RH blood type patient is in an emergency, if a negative blood type is in short supply, a small amount of weak positive blood types can be input for the patient, the life of the patient is temporarily kept, a certain time is won for the patient, and the patient waits for the negative blood type.

Referring to fig. 3, aiming at the detection method, the invention further provides a full-automatic RH system blood type detector, which comprises a sample plate 1, a vibrator 2, a micro-porous plate 3, an antibody 4, a deep-hole plate 5, a centrifuge 6 and a centrifuge 7.

The blood type detector adopts the method for detection, and generally takes 3-8 min; the detection time is short, and the efficiency is high.

In order to verify the detection method, the invention is provided with the following embodiments, and the blood type pictures detected in each embodiment are counted to verify the detection precision of the method. Since the photo interpretation process of each example results in several pictures, the invention only lists one group of pictures in example 1 as reference, see fig. 2.

Example 1

Step one blood sample pretreatment

Step one, blood sample is put on machine

Step two, adding the antibody into the microporous plate

The microplate 3 is now placed on the oscillator 2, and the five antibodies 4 are then added to the microplate 3, respectively. The microplate 3 is a 96-well plate, and the volume of each well in the microplate 3 is 120 ul. The addition amount of the five antibodies 4 in the microplate 3 was 13ul, respectively.

Step three blood sample dilution

Dripping normal saline into the deep hole plate 5, transferring the red blood cell suspension in the blood test tube through a control system of the blood type detector, and adding the red blood cell suspension into the deep hole plate 5 in which the normal saline is dripped; diluting the erythrocyte suspension by using normal saline to obtain a diluent.

In the dilution, the volume of the erythrocyte suspension was 2.4%.

Step four antigen antibody mixing

And adding the diluent into the microporous plate 3 to which the antibody 4 is added dropwise, and mixing the diluent with the five antibodies 4 respectively, wherein the addition amount of the diluent in the microporous plate 3 is 25 ul. The control system of the blood type detector controls the oscillator 2 to work, so that the diluent and the antibody 4 are uniformly mixed, the oscillation time of the oscillator is 26s, the amplitude is 1.5mm, and the frequency is 679 RPM.

Step five centrifugation

And (3) putting the vibrated microporous plate 3 in the step four into a centrifuge 6 of a blood type detector, and centrifuging the fully mixed antigen antibody 4. In the process of centrifugation, the relative centrifugal force of the centrifuge 6 is 271g, and the centrifugation time is 0.9 min.

Step six oscillation dispersion

Step seven photographing interpretation

In weakly positive blood types, incomplete clots can be seen.

Example 2

Step one, blood sample is put on machine

Step two, adding the antibody into the microporous plate

The microplate 3 is now placed on the oscillator 2, and the five antibodies 4 are then added to the microplate 3, respectively. The microplate 3 is a 96-well plate, and the volume of each well in the microplate 3 is 120 ul. The addition amount of the five antibodies 4 in the microplate 3 was 32ul, respectively.

Step three blood sample dilution

Dripping normal saline into the deep hole plate 5, transferring the red blood cell suspension in the blood test tube through a control system of the blood type detector, and adding the red blood cell suspension into the deep hole plate 5 in which the normal saline is dripped; diluting the erythrocyte suspension by using normal saline to obtain a diluent.

In the dilution, the volume of the erythrocyte suspension was 3.4%.

Step four antigen antibody mixing

And adding the diluent into the microporous plate 3 to which the antibody 4 is added dropwise, and mixing the diluent with the five antibodies 4 respectively, wherein the addition amount of the diluent in the microporous plate 3 is 32 ul. The control system through the blood type detector controls oscillator 2 to work, makes diluent and antibody 4 misce bene, and the oscillation time of oscillator is 29s, and amplitude 2mm, frequency 2100 RPM.

Step five centrifugation

And (3) putting the vibrated microporous plate 3 in the step four into a centrifuge 6 of a blood type detector, and centrifuging the fully mixed antigen antibody 4. During the centrifugation, the relative centrifugal force of the centrifuge 6 was 410g, and the centrifugation time was 1 min.

Step six oscillation dispersion

Step seven photographing interpretation

In weakly positive blood types, incomplete clots can be seen.

Example 3

Step one, blood sample is put on machine

Step two, adding the antibody into the microporous plate

The microplate 3 is now placed on the oscillator 2, and the five antibodies 4 are then added to the microplate 3, respectively. The microplate 3 is a 96-well plate, and the volume of each well in the microplate 3 is 120 ul. The addition amount of the five antibodies 4 in the microplate 3 was 48ul, respectively.

Step three blood sample dilution

Dripping normal saline into the deep hole plate 5, transferring the red blood cell suspension in the blood test tube through a control system of the blood type detector, and adding the red blood cell suspension into the deep hole plate 5 in which the normal saline is dripped; diluting the erythrocyte suspension by using normal saline to obtain a diluent.

In the dilution, the volume of the red blood cell suspension was 3.7%.

Step four antigen antibody mixing

The diluent is added into the micropore plate 3 to which the antibody 4 is already dripped, the diluent and the five antibodies 4 are respectively mixed, and the addition amount of the diluent in the micropore plate 3 is 47 ul. The control system through the blood type detector controls 2 work of oscillators, makes diluent and antibody 4 misce bene, and the oscillation time of oscillator is 35s, and amplitude 3mm, frequency 2640 RPM.

Step five centrifugation

And (3) putting the vibrated microporous plate 3 in the step four into a centrifuge 6 of a blood type detector, and centrifuging the fully mixed antigen antibody 4. In the centrifugation process, the relative centrifugal force of the centrifuge 6 is 473g, and the centrifugation time is 1.3 min.

Step six oscillation dispersion

Step seven photographing interpretation

In weakly positive blood types, incomplete clots can be seen.

In summary, in the fully automatic blood type detector, the mixture of the antigen and the antibody is first subjected to vibration mixing, so that the mixing uniformity of the antigen and the antibody can be improved, the mixture of the antigen and the antibody is subjected to centrifugal operation after being uniformly mixed, positive and weak positive samples are agglutinated under the action of centrifugal force, and then vibration is performed, so that the agglutination of the weak positive samples is vibrated away, and the agglutination of the positive samples is still gathered together, so that the positive and the weak positive blood can be distinguished. In conclusion, the invention can realize full-automatic detection of blood, does not need manual operation, has short detection time and high efficiency, can accurately judge the blood types of positive and weak positive, and avoids the phenomenon that the transfusion of a patient is delayed due to the fact that the blood type of the weak positive cannot be detected, so that the patient cannot be cured.

The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof, and it should be understood that various changes and modifications can be effected therein by one skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.