CN107300622B - Prediction method of amniotic fluid embolism coagulation dysfunction - Google Patents
Prediction method of amniotic fluid embolism coagulation dysfunction Download PDFInfo
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/22—Haematology
- G01N2800/224—Haemostasis or coagulation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/36—Gynecology or obstetrics
- G01N2800/368—Pregnancy complicated by disease or abnormalities of pregnancy, e.g. preeclampsia, preterm labour
Abstract
The invention relates to a prediction method of amniotic fluid embolism coagulation disorder, which adopts a thromboelastogram capable of reflecting the coagulation process and fibrinolysis holomorphy to induce whole blood of a pregnant woman in vitro by adopting amniotic fluid and urokinase with proper concentration, so as to stimulate the blood to generate high coagulation and fibrinolysis processes, evaluate the sensitivity and tolerance of the pregnant woman to the amniotic fluid and urokinase through the change of indexes of the thromboelastogram, and predict the possibility of the occurrence of the coagulation disorder caused by the amniotic fluid; when LY30 value is not less than 7.5, the incidence rate of amniotic fluid embolism coagulation dysfunction of pregnant women is high, and the amniotic fluid embolism is screened and taken measures in advance to prevent the occurrence of the amniotic fluid embolism.
Description
Technical Field
The invention belongs to the technical field of blood coagulation detection, and particularly relates to a prediction method of amniotic fluid embolism blood coagulation dysfunction.
Background
Amniotic fluid embolism is a pregnancy anaphylaxis syndrome, and patients usually show anaphylactic shock and acute pulmonary embolism. It is also considered by researchers that amniotic fluid embolism is a consumptive coagulopathy, and patients show a large amount of consumption of blood coagulation substances and hyperfibrinolysis, and the whole body of puerpera bleeds, so that the puerpera bleeds and dies. At present, no test method for prevention and detection exists, and the establishment of a sensitive and reliable test method has important significance if high risk groups with amniotic fluid embolism and blood coagulation dysfunction can be predicted and screened prenatally. Miriam J.P.Harnet adopts thromboelastogram to observe the influence of amniotic fluid on the blood coagulation function of pregnant women in vitro, and researches suggest that the amniotic fluid further stimulates the occurrence of hypercoagulation, but experiments do not realize the change of fibrinolysis indexes.
Thromboelastography (TEG) is mainly used clinically for the detection of blood coagulation function in whole blood samples. The thromboelastogram sample is a whole blood sample of a patient, the whole blood sample of the patient is placed in a sample cup during detection, a detection rod connected with a motion sensing and conducting system is inserted into the sample cup, and the sample is activated by a kaolin solution at a constant temperature of 37 ℃ and then detected. The sample cup rotates at a constant speed of 4 degrees and 45 minutes, and the detection rod senses the resistance of the sample to move and records the resistance to generate a trace graph and parameters. The trace is roughly divided into two parts, the coagulation phase and the fibrinolysis phase. In the coagulation phase, the time R is the time required from the start of the assay to the rise in the amplitude of the curve to 2mm, corresponding to the process in which the gradual activation of various coagulation factors leads to the initial formation of fibrin. The K time is the time taken for the amplitude of the curve to rise from 2mm to 20mm, corresponding to fibrin cross-linking and its interaction with platelets. The angle A is the included angle between the tangent line of the maximum curve radian of the trace and the horizontal line, reflects the interaction between fibrin and platelets together with the K time, and becomes an important substitute index when the severe low coagulation K value of a patient cannot be measured. The Maximum Amplitude (MA) is the maximum trace amplitude, corresponding to the maximum intensity of the clot, and is related to platelet concentration, platelet function, and platelet-fibrin interaction. The clot enters the fibrinolytic stage after it reaches maximum intensity, with the more common indicators being Ly30 and Ly60, which represent the percentage of thrombolysis 30 minutes and 60 minutes after the trace reaches MA, respectively, as calculated from the decreasing area under the curve, reflecting the extent to which plasmin dissolves the thrombus.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a prediction method of the coagulation dysfunction of the amniotic fluid embolism. The method is based on the thromboelastogram, adopts the thromboelastogram which can reflect the blood coagulation process and the fibrinolysis physiognomy, carries out the in-vitro induction of amniotic fluid and urokinase on the whole blood of the pregnant woman, stimulates the blood to generate hypercoagulation (blood coagulation factor consumption) and fibrinolysis process (urokinase induction), evaluates the sensitivity and the tolerance of the pregnant woman to the amniotic fluid and the urokinase through the change of the indexes of the thromboelastogram, predicts the possibility of the occurrence of the coagulation disorder caused by the amniotic fluid, and screens and prevents the occurrence of the amniotic fluid embolism in advance.
The technical scheme adopted by the invention is as follows:
the application of the thromboelastogram in predicting the coagulation dysfunction of the amniotic fluid embolism.
A method for predicting amniotic fluid embolism coagulation dysfunction based on thromboelastography comprises the following steps:
(1) fully activating a blood sample by using a kaolin solution to obtain an activated blood sample;
(2) adding the activated blood sample into a sample cup containing calcium chloride solution, and slowly adding amniotic fluid and urokinase into the sample cup to obtain a blood sample to be detected; the volume ratio of the calcium chloride solution, the amniotic fluid, the urokinase and the blood sample after activation is 15-25:5-30:5-30: 300-400;
(3) and (3) placing the blood sample to be tested in a thrombelastogram instrument for testing to obtain a fibrinolysis index LY30 value.
The activity of the urokinase is 20-200U/ml.
In the step (1), the kaolin solution is fully mixed with the blood sample to activate the blood sample, and the activation time is 3-6 min.
In the step (1), the concentration of the kaolin solution is 0.2-0.5 mg/ml.
The volume ratio of the kaolin solution to the blood sample is 5:400-15: 300.
In the step (2), the adding amount of the amniotic fluid in the sample cup is 5-30 μ l.
In the step (2), the adding amount of the urokinase in the sample cup is 5-30 μ l.
In the step (2), the concentration of the calcium chloride solution in the sample cup is 0.1-0.5 mol/L.
When LY30 value ≧ 7.5, the incidence of amniotic fluid embolism blood coagulation dysfunction is high.
The invention has the beneficial effects that:
the prediction method of the amniotic fluid embolism coagulation disorder adopts the thromboelastogram which can reflect the coagulation process and the fibrinolysis holomorphy to carry out in-vitro induction on whole blood of a pregnant woman by adopting the amniotic fluid and the urokinase with proper concentrations, stimulates the blood to have high coagulation (blood coagulation factor consumption) and fibrinolysis process (urokinase induction), evaluates the sensitivity and the tolerance of the pregnant woman to the amniotic fluid and the urokinase through the change of indexes of the thromboelastogram, and predicts the possibility of the occurrence of the coagulation disorder caused by the amniotic fluid; when LY30 value is not less than 7.5, the incidence rate of amniotic fluid embolism coagulation dysfunction of pregnant women is high, and the amniotic fluid embolism is screened and taken measures in advance to prevent the occurrence of the amniotic fluid embolism.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention.
Example 1
The embodiment provides a method for predicting amniotic fluid embolism coagulation dysfunction based on thromboelastography, which comprises the following steps:
(1) fully mixing 5 mul of kaolin solution with the concentration of 0.5mg/ml with 300 mul of blood sample to activate the blood sample, wherein the activation time is 3min, and obtaining the activated blood sample;
(2) adding the activated blood sample into a sample cup containing 15 mu L of calcium chloride solution with the concentration of 0.5mol/L, and slowly adding 5 mu L of amniotic fluid and 5 mu L of urokinase (the enzyme activity of the urokinase is 20U/ml) into the sample cup to obtain a blood sample to be detected;
(3) and (3) placing the blood sample to be tested in a thromboelastography instrument for testing to obtain a fibrinolysis index LY30 value, wherein when the LY30 value is not less than 7.5, the incidence rate of the amniotic fluid embolism coagulation dysfunction of the pregnant woman is high, and intervention measures should be taken in advance.
Example 2
The embodiment provides a method for predicting amniotic fluid embolism coagulation dysfunction based on thromboelastography, which comprises the following steps:
(1) fully mixing 15 mu l of kaolin solution with the concentration of 0.2mg/ml with 400 mu l of blood sample to activate the blood sample, wherein the activation time is 6min, and obtaining the activated blood sample;
(2) adding the activated blood sample into a sample cup containing 25 mu L of calcium chloride solution with the concentration of 0.1mol/L, and slowly adding 30 mu L of amniotic fluid and 30 mu L of urokinase (the enzyme activity of the urokinase is 200U/ml) into the sample cup to obtain a blood sample to be detected;
(3) and (3) placing the blood sample to be tested in a thromboelastography instrument for testing to obtain a fibrinolysis index LY30 value, wherein when the LY30 value is not less than 7.5, the incidence rate of the amniotic fluid embolism coagulation dysfunction of the pregnant woman is high, and intervention measures should be taken in advance.
Example 3
The embodiment provides a method for predicting amniotic fluid embolism coagulation dysfunction based on thromboelastography, which comprises the following steps:
(1) fully mixing 10 mu l of kaolin solution with the concentration of 0.4mg/ml with 340 mu l of blood sample to activate the blood sample, wherein the activation time is 4min, and obtaining the activated blood sample;
(2) adding the activated blood sample into a sample cup containing 20 mu L of calcium chloride solution with the concentration of 0.3mol/L, and slowly adding 10 mu L of amniotic fluid and 10 mu L of urokinase (the enzyme activity of the urokinase is 58U/ml) into the sample cup to obtain a blood sample to be detected;
(3) and (3) placing the blood sample to be tested in a thromboelastography instrument for testing, wherein when the LY30 value is not less than 7.5, the incidence rate of the amniotic fluid embolism and blood coagulation dysfunction of the pregnant woman is high, and an intervention measure is required to be taken in advance.
Example 4
The embodiment provides a method for predicting amniotic fluid embolism coagulation dysfunction based on thromboelastography, which comprises the following steps:
(1) fully mixing 10 mu l of kaolin solution with the concentration of 0.2mg/ml with 340 mu l of blood sample to activate the blood sample, wherein the activation time is 5min, and obtaining the activated blood sample;
(2) adding the activated blood sample into a sample cup containing 20 mu L of calcium chloride solution with the concentration of 0.3mol/L, and slowly adding 10 mu L of amniotic fluid and 15 mu L of urokinase (the enzyme activity of the urokinase is 85U/ml) into the sample cup to obtain a blood sample to be detected;
(3) and (3) placing the blood sample to be tested in a thromboelastography instrument for testing, wherein when the LY30 value is not less than 7.5, the incidence rate of the amniotic fluid embolism and blood coagulation dysfunction of the pregnant woman is high, and an intervention measure is required to be taken in advance.
Example 5
The embodiment provides a method for predicting amniotic fluid embolism coagulation dysfunction based on thromboelastography, which comprises the following steps:
(1) fully mixing 10 mu L of kaolin solution with the concentration of 0.3mol/L with 340 mu L of blood sample to activate the blood sample, wherein the activation time is 5min, and obtaining the activated blood sample;
(2) adding the activated blood sample into a sample cup containing 20 mu L of calcium chloride solution with the concentration of 0.2mol/L, and slowly adding 10 mu L of amniotic fluid and 20 mu L of urokinase (the enzyme activity of the urokinase is 120U/ml) into the sample cup to obtain a blood sample to be detected;
(3) and (3) placing the blood sample to be tested in a thromboelastography instrument for testing, wherein when the LY30 value is not less than 7.5, the incidence rate of the amniotic fluid embolism and blood coagulation dysfunction of the pregnant woman is high, and an intervention measure is required to be taken in advance.
Example 6
The embodiment provides a method for predicting amniotic fluid embolism coagulation dysfunction based on thromboelastography, which comprises the following steps:
(1) fully mixing 10 mu L of kaolin solution with the concentration of 0.4mol/L with 340 mu L of blood sample to activate the blood sample, wherein the activation time is 5min, and obtaining the activated blood sample;
(2) adding the activated blood sample into a sample cup containing 20 mu L of calcium chloride solution with the concentration of 0.2mol/L, and slowly adding 10 mu L of amniotic fluid and 25 mu L of urokinase (the enzyme activity of the urokinase is 150U/ml) into the sample cup to obtain a blood sample to be detected;
(3) and (3) placing the blood sample to be tested in a thromboelastography instrument for testing, wherein when the LY30 value is not less than 7.5, the incidence rate of the amniotic fluid embolism and blood coagulation dysfunction of the pregnant woman is high, and an intervention measure is required to be taken in advance.
Examples of the experiments
The test method described in example 3 was used to perform a confirmatory experiment on a clinical application test group (75 pregnant women) and a clinical application control group (32 non-pregnant women). The results are shown in Table 1.
TABLE 1 LY30 Difference before and after Induction with amniotic fluid and urokinase
As can be seen from Table 1, before induction, the number of LY30 ≧ 7.5 (high incidence of amniotic fluid embolism coagulation dysfunction) in each of the experimental group and the control group was 1, and the ratio of difference p between the samples of the experimental group and the control group was 0.531, and χ20.393, indicating that the difference between the experimental group and the control group before induction is not significant; after the induction is carried out by adopting 10 mu l of amniotic fluid and 10 mu l of urokinase, the number of people with LY30 being more than or equal to 7.5 (the occurrence rate of the amniotic fluid embolism and blood coagulation dysfunction is high) in an experimental group is 22, the number of people with LY30 being more than or equal to 7.5 (the occurrence rate of the amniotic fluid embolism and blood coagulation dysfunction is high) in a control group is 31, and the difference ratio p between samples of the experimental group and the control group is 0.001 and chi is240.933, indicating that the difference between the experimental group and the control group after induction is very significant. Therefore, the method can be used for screening the pregnant women with higher incidence rate of the amniotic fluid embolism blood coagulation dysfunction so as to take intervention measures in advance.
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and all the changes or substitutions should be covered within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (1)
1. A construction method of a prediction model of amniotic fluid embolism coagulation dysfunction based on thromboelastography is characterized by comprising the following steps:
(1) fully mixing 10 mu l of kaolin solution with the concentration of 0.4mg/ml with 340 mu l of blood sample to activate the blood sample, wherein the activation time is 4min, and obtaining the activated blood sample;
(2) adding the activated blood sample into a sample cup containing 20 mul of calcium chloride solution with the concentration of 0.3mol/L, and slowly adding 10 mul of amniotic fluid and 10 mul of urokinase into the sample cup, wherein the enzymatic activity of the urokinase is 58U/ml, so as to obtain a blood sample to be detected;
(3) and (3) placing the blood sample to be tested in a thromboelastography instrument for testing to obtain a fibrinolysis index LY30 value, namely constructing a prediction model of the amniotic fluid embolism coagulation dysfunction, wherein when the LY30 value is not less than 7.5, the incidence rate of the amniotic fluid embolism coagulation dysfunction of the pregnant woman is high, and intervention measures should be taken in advance.
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Citations (3)
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| CN101587063A (en) * | 2009-06-19 | 2009-11-25 | 西安交通大学 | Method for detecting hemostatic capability |
| CN104661582A (en) * | 2012-05-14 | 2015-05-27 | 高斯外科公司 | System and methods for managing blood loss of a patient |
| CN106546743A (en) * | 2016-11-04 | 2017-03-29 | 中山大学 | A kind of real-time monitoring system and monitoring method of amniotic embolism serological index |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101587063A (en) * | 2009-06-19 | 2009-11-25 | 西安交通大学 | Method for detecting hemostatic capability |
| CN104661582A (en) * | 2012-05-14 | 2015-05-27 | 高斯外科公司 | System and methods for managing blood loss of a patient |
| CN106546743A (en) * | 2016-11-04 | 2017-03-29 | 中山大学 | A kind of real-time monitoring system and monitoring method of amniotic embolism serological index |
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