CN109406461B - Method for judging platelet aggregation rate measurement result and method for calibrating platelet aggregation instrument - Google Patents
Method for judging platelet aggregation rate measurement result and method for calibrating platelet aggregation instrument Download PDFInfo
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Abstract
The invention provides a method for judging a platelet aggregation rate measurement result, which comprises the following steps: selecting a normal whole blood sample, measuring the platelet poor plasma transmittance PPP0 and the platelet rich plasma transmittance PRP0, and calculating the transmittance difference P0 of the whole blood sample to be PPP0-PRP 0; selecting a whole blood sample to be detected, measuring the transmittance PPP and the transmittance PRP of platelet-poor plasma, and calculating the transmittance difference P of the sample to be detected as PPP-PRP; calculating the platelet separation index DI ═ P/P0 ═ (PPP-PRP)/(PPP0-PRP0) of the tested sample; measuring the platelet aggregation rate of the platelet-rich plasma by using a platelet aggregation instrument; and judging whether the measured platelet aggregation rate is reliable or not according to the platelet separation index DI of the tested sample. The invention can reliably judge the measurement result of the platelet aggregation rate of the tested sample by referring to the platelet separation degree index of the tested sample.
Description
Technical Field
The invention relates to the technical field of medical treatment, in particular to a method for judging a platelet aggregation rate measurement result and a calibration method suitable for a platelet aggregation instrument.
Background
The optical turbidimetry is a practical method which is easy to popularize in clinic and is used for detecting the platelet aggregation function, the method is simple to operate, low in price and good in correlation with clinical events, and the method is a gold standard for detecting the platelet aggregation function.
Platelet Rich Plasma (PRP) is a suspension of platelets in plasma, the platelets are uniformly dispersed, the turbidity is in direct proportion to the number of platelets contained in the plasma, after a platelet aggregation inducer is added, the inducer is combined with a receptor on the platelets under the condition of stirring, the platelets are activated, the platelets deform to further release endogenous substances, aggregation is caused, the transmittance is increased, the higher the degree of platelet aggregation in the PRP is, the lower the turbidity is, the higher the transmittance is, optical signals are converted into electric signals, and a platelet aggregation instrument automatically carries out dynamic measurement and recording, so that a platelet aggregation curve is drawn. When actually calculating the platelet aggregation rate, assuming that the transmittance of Platelet Poor Plasma (PPP) is PPP, which indicates the light flux when all platelets aggregate, i.e., the aggregation rate is 100%, assuming that the transmittance of Platelet Rich Plasma (PRP) is PRP, which indicates the light flux when platelets do not aggregate, i.e., the aggregation rate is 0%, adding an inducer to PRP to cause aggregation reaction, assuming that the light flux value at time i is r (i), the platelet aggregation rate calculation formula is as follows:
platelet aggregation rate = MAX ((r (i) -PRP)/(PPP-PRP)).
The platelet aggregation rate measured by the existing method is stable after the platelet concentration in a blood sample reaches a certain degree, but if the platelet concentration is less than a certain value, the aggregation rate is found to be reduced, so that the aggregation rate reported by an instrument is unreliable, and the platelet aggregation function cannot be accurately reflected. The method is complicated to operate, and needs extra instruments such as a blood analyzer and the like, so that a result cannot be obtained at one time when the platelet aggregation rate is measured.
Disclosure of Invention
In view of the above, it is a main object of the present invention to provide a method for determining a result of measuring a platelet aggregation rate, which is simple in operation and capable of more reliably determining a result of measuring a platelet aggregation rate by comparing a difference in transmittance of a platelet poor plasma in a sample to be measured with a difference in transmittance of a platelet poor plasma in a normal sample without using an extra blood analyzer or the like to evaluate a platelet concentration in the sample.
The technical scheme adopted by the invention is that the method for judging the platelet aggregation rate measurement result comprises the following steps:
A. selecting a normal whole blood sample, separating platelet poor plasma and platelet rich plasma according to a standard operation procedure, respectively measuring the transmittance of the platelet poor plasma PPP0 and the transmittance of the platelet rich plasma PRP0, and then calculating the transmittance difference P0= PPP0-PRP0 of the normal whole blood sample;
B. selecting a whole blood sample to be detected, separating platelet poor plasma and platelet rich plasma, measuring the transmittance PPP and the transmittance PRP of the platelet poor plasma, and calculating the transmittance difference P = PPP-PRP of the whole blood sample to be detected;
C. calculating a platelet separation index DI = P/P0= (PPP-PRP)/(PPP0-PRP0) of the measured whole blood sample from a ratio of a difference P in transmittance of the measured whole blood sample and a difference P0 in transmittance of the normal whole blood sample;
D. selecting platelet-rich plasma in a whole blood sample to be detected, adding a platelet inducer, mixing and stirring, and measuring the platelet aggregation rate of the platelet-rich plasma by using a platelet aggregation instrument;
E. and determining the reliability of the platelet aggregation rate of the platelet rich plasma measured by the platelet aggregation instrument according to the calculated platelet separation index DI of the whole blood sample to be measured.
Therefore, the platelet separation degree parameter of the detected sample is obtained by comparing the difference of the light transmittance of the poor platelet rich in the detected sample with the light transmittance of the platelet rich in the normal sample, so that the platelet aggregation rate of the platelet rich plasma in the measured sample is effectively judged, and whether the measured platelet aggregation function is a correct result is obtained. Compared with the traditional method, the platelet aggregation function detection method has the advantages that a blood analyzer is additionally introduced to count platelets before platelet aggregation function detection is carried out, the platelet aggregation rate result is judged according to the platelet concentration obtained by platelet counting, equipment resources are saved, and the test flow is simplified.
Wherein the step E comprises:
if the DI is more than or equal to 1, the result of the measured platelet aggregation rate is judged to be reliable;
if the DI is not less than 1/3 and less than 1, the result of the measured platelet aggregation rate is judged to be reliable after calibration;
if DI < 1/3, the platelet concentration of the whole blood sample is too low, and the result of the platelet aggregation rate is determined to be suspicious.
Therefore, whether the result of the measured platelet aggregation rate is reliable or not can be quickly judged according to the calculated platelet separation index of the measured sample, if the DI index defined above is more than 1, the difference of the poor and rich platelet concentration of the sample is more than that of a normal sample which is normally separated, in principle, the situation has no adverse effect on the detection result, and the result of the platelet aggregation rate obtained by the measurement of an instrument can be directly reported; if the DI index is less than 1 and greater than 1/3, i.e. the difference between the lean and rich of the sample is 1/3 or more of that of the normal sample, the aggregation result can be calibrated by a calibration function, and the calibrated result is a reportable platelet aggregation rate result; if the DI index is less than 1/3, it should be suggested that the sample platelet concentration is too low and the result of the platelet aggregation rate is questionable, and the result of the measured platelet aggregation rate cannot accurately and reliably reflect the platelet aggregation function.
In a further refinement, when the calculated platelet separation index DI is 1/3 ≦ DI < 1, the calibrating comprises:
the measured platelet aggregation rate results are calibrated by a mapping function comprising a linear, polynomial or other form of function, the calibration aimed at minimizing the error between the measured results and those measured at DI ≧ 1.
From the above, it is proved by experimental data that when the platelet separation index DI is gradually adjusted downward from 1, the measured platelet aggregation rate is correspondingly decreased, and the decrease trend continues until the platelet separation index is 1/3, so that the measured result of the platelet aggregation rate can be calibrated by using a mapping function to obtain an expected normal value, whereas when the platelet separation index DI is continuously adjusted downward, the platelet aggregation rate is greatly decreased, and at this time, the decrease trend does not meet the decrease trend reflected by the mapping function, and a normal value cannot be obtained by means of function calibration, so that the measured result of the platelet aggregation rate needs to be reported as an in-doubt result, which indicates that the result cannot directly reflect the platelet aggregation function.
The invention also provides a calibration method of the platelet aggregation instrument, which comprises the following steps: the method for determining the result of measurement of platelet aggregation rate;
and calibrating whether the measurement result of the platelet aggregation instrument is reliable or not according to the accuracy of the platelet aggregation rate determined by the platelet aggregation rate measurement result determination method.
Therefore, the aggregation rate measuring method can be applied to most of the existing platelet aggregation instruments, and the function test of the existing platelet aggregation instruments is realized so as to verify whether the measurement results of the aggregation instruments are reliable or not.
Drawings
Fig. 1 is a flowchart of the method for determining the result of measurement of the platelet aggregation rate according to the present invention.
FIG. 2 is a schematic diagram showing platelet aggregation of the platelet rich plasma of the test sample.
Detailed Description
The main object of the present invention is to provide a method for determining a result of measuring a platelet aggregation rate, which realizes detection and calibration of a platelet concentration by referring to a platelet separation degree parameter when separating platelets, thereby determining a measurement result more accurately and reliably when measuring a platelet aggregation rate.
The present invention will be described in detail below with reference to fig. 1.
As shown in the flowchart of the method for determining a result of measuring a platelet aggregation rate in fig. 1, the present invention provides a method for determining a result of measuring a platelet aggregation rate, comprising the steps of:
s01: selecting a normal whole blood sample, separating platelet poor plasma and platelet rich plasma according to a standard operation procedure, respectively measuring the transmittance of the platelet poor plasma PPP0 and the transmittance of the platelet rich plasma PRP0, and calculating the transmittance difference P0= PPP0-PRP0 of the normal whole blood sample;
in step S01, a blood collection test tube is used to collect a normal whole blood sample, the whole blood sample is subjected to anticoagulation treatment to generate anticoagulated whole blood, the anticoagulated whole blood is centrifuged at a low centrifugation speed (usually 500 rpm-1000 rpm) for five minutes, the supernatant is taken to prepare a platelet-rich plasma sample, the residual blood is centrifuged at a high centrifugation speed (usually 2000 rpm-3000 rpm) for ten minutes, and the supernatant is taken to prepare a platelet-poor plasma sample;
measuring the concentrations of the platelet poor plasma and the platelet rich plasma by using a blood cell counter, storing the platelet rich plasma sample into a platelet rich plasma sample cup of a sample disc device after confirming that the concentrations meet the centrifugal standard, and measuring to obtain the transmittance PRP0 of the platelet rich plasma; and (4) storing the platelet poor plasma sample in a platelet poor plasma sample cup of the sample plate device, and measuring to obtain the platelet poor plasma transmittance PPP 0.
S02: selecting a whole blood sample to be detected, separating platelet poor plasma and platelet rich plasma, measuring the transmittance PPP and the transmittance PRP of the platelet poor plasma, and calculating the transmittance difference P = PPP-PRP of the whole blood sample to be detected;
in step S02, another blood collection test tube is used to collect a whole blood sample to be tested, the whole blood sample is subjected to anticoagulation treatment to generate anticoagulated whole blood, the anticoagulated whole blood is centrifuged at a low centrifugation speed (usually 500 rpm-1000 rpm) for five minutes, the supernatant is taken to prepare a platelet-rich plasma sample, the residual blood is centrifuged at a high centrifugation speed (usually 2000 rpm-3000 rpm) for ten minutes, and the supernatant is taken to prepare a platelet-poor plasma sample;
storing the platelet-rich plasma sample in a platelet-rich plasma sample cup of a sample disc device, and measuring to obtain the platelet-rich plasma transmittance PRP; and (4) storing the platelet poor plasma sample into a platelet poor plasma sample cup of the sample plate device, and measuring to obtain the platelet poor plasma transmittance PPP.
S03: calculating a platelet separation index DI = P/P0= (PPP-PRP)/(PPP0-PRP0) of the measured whole blood sample from a ratio of a difference P in transmittance of the measured whole blood sample and a difference P0 in transmittance of the normal whole blood sample;
s04: selecting platelet-rich plasma in a whole blood sample to be detected, adding a platelet inducer, mixing and stirring, and measuring the platelet aggregation rate of the platelet-rich plasma by using a platelet aggregation instrument;
as shown in fig. 2, after the whole blood sample to be tested is centrifuged, the transmittance PPP of platelet poor plasma is 100%, and the transmittance PRP of platelet rich plasma is 0%, at this time, a certain amount of platelet rich plasma is added into a new test tube, a platelet inducer and a magnetic stirrer are added for stirring, the platelet inducer is combined with a receptor on the platelets in the platelet rich plasma, the platelets are activated, deformation occurs to further release endogenous substances, aggregation is caused, the transmittance is increased, at this time, optical signals are converted into electrical signals, and a platelet aggregation instrument performs dynamic measurement and recording, thereby depicting the aggregation curve of the platelets.
S05: and (3) judging whether the platelet aggregation rate of the platelet rich plasma measured by a platelet aggregation instrument is reliable or not according to the calculated platelet separation index DI of the measured whole blood sample:
if the DI is more than or equal to 1, judging that the result of the measured platelet aggregation rate is a correct value;
if 1/3 is less than or equal to DI < 1, the measured platelet aggregation rate result is determined to be a correct value after calibration;
if DI < 1/3, the platelet concentration of the whole blood sample is too low, and the measured platelet aggregation rate is determined as a suspected value;
in step S05, the result of the platelet aggregation rate of the platelet-rich plasma in the whole blood sample to be tested measured by the platelet aggregation meter is determined according to the calculated platelet separation index:
if the platelet separation index DI is larger than 1, it indicates that the difference between the poor and rich platelet concentrations of the tested sample is larger than that of the normal sample separated normally, and in principle, such a situation should not have any adverse effect on the detection result, and the platelet aggregation rate obtained by the measurement of the instrument can be directly reported to generate the detection result.
If the platelet separation index DI is less than 1 and greater than 1/3, i.e. the difference between the poverty and richness of the sample is 1/3 or more of that of the normal sample, the aggregation result can be calibrated by a mapping function, the mapping function includes a linear function, a polynomial function or other forms of functions, and the result after calibration by the mapping function is the result of the platelet aggregation rate that can generate a detection report;
based on the above platelet separation index, a large number of related experiments are performed, and the experimental results prove that when the platelet separation index DI is gradually adjusted from 1 to downward, the measured platelet aggregation rate is correspondingly decreased, and the decrease trend continues to 1/3, and the decrease trend can be summarized as a mapping function formula according to the experimental results, and the specific steps are as follows:
(1): taking a blood sample of a normal human, and measuring the platelet aggregation rate to be R0;
(2): performing contrast dilution on the blood sample by using normal saline, obtaining a sample with the platelet concentration being half of the original sample, namely the separation degree being 1/2, and performing platelet aggregation rate detection to obtain the aggregation rate being R1;
(3): the calibration factor K, K is also 1 in case of a degree of separation of 1, K is R0/R1 in case of a degree of separation of 1/2;
(4): therefore, assuming that K = a × D + b and D is the actually measured platelet aggregation rate, the relationship function between the separation degree and the calibration function is substituted with the calibration coefficients corresponding to the separation degree of 1 and the separation degree of 1/2, so as to obtain two parameters, i.e., a =2 × 2 (1-R0/R1) and b =2 × R0/R1-1;
therefore, the measured platelet aggregation rate results may be calibrated using the mapping function described above to obtain the desired normal value.
If the platelet separation index DI is less than 1/3, the user should be prompted that the sample platelet concentration is too low, the result of the platelet aggregation rate is questioned, and the result of the measured platelet aggregation rate cannot visually reflect the aggregation function of the platelets;
similarly, in the above experiment, when the platelet separation index DI is smaller than 1/3 and the adjustment is continued, the platelet aggregation rate will be greatly decreased, and at this time, the trend reflected by the mapping function is not met, and a normal value cannot be obtained by means of function calibration, so that the measured platelet aggregation rate needs to be reported as an in-doubt result, which indicates that the result cannot intuitively reflect the platelet aggregation function.
Based on the method for judging the platelet aggregation rate measurement result, the invention also provides a method for calibrating the platelet aggregation instrument, which is suitable for most of the existing platelet aggregation instruments;
and calibrating whether the measurement result of the platelet aggregation instrument is reliable or not according to the accuracy of the platelet aggregation rate determined by the platelet aggregation rate measurement result determination method, so as to realize the function test of the conventional platelet aggregation instrument and verify the measurement reliability of the platelet aggregation instrument.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (3)
1. A method for determining a result of measurement of a platelet aggregation rate, comprising the steps of:
A. selecting a normal whole blood sample, separating platelet poor plasma and platelet rich plasma according to a standard operation procedure, respectively measuring the transmittance of the platelet poor plasma PPP0 and the transmittance of the platelet rich plasma PRP0, and then calculating the transmittance difference P0= PPP0-PRP0 of the normal whole blood sample;
B. selecting a whole blood sample to be detected, separating platelet poor plasma and platelet rich plasma, measuring the transmittance PPP and the transmittance PRP of the platelet poor plasma, and calculating the transmittance difference P = PPP-PRP of the whole blood sample to be detected;
C. calculating a platelet separation index DI = P/P0= (PPP-PRP)/(PPP0-PRP0) of the measured whole blood sample from a ratio of a difference P in transmittance of the measured whole blood sample and a difference P0 in transmittance of the normal whole blood sample;
D. selecting platelet-rich plasma in a whole blood sample to be detected, adding a platelet inducer, mixing and stirring, and measuring the platelet aggregation rate of the platelet-rich plasma by using a platelet aggregation instrument;
E. and (3) judging the reliability of the platelet aggregation rate of the platelet rich plasma measured by a platelet aggregation instrument according to the calculated platelet separation index DI of the whole blood sample to be measured:
if the DI is more than or equal to 1, the result of the measured platelet aggregation rate is judged to be reliable;
if the DI is not less than 1/3 and less than 1, the result of the measured platelet aggregation rate is judged to be reliable after calibration;
if DI < 1/3, the platelet concentration of the whole blood sample is too low, and the result of the platelet aggregation rate is determined to be suspicious.
2. The method of claim 1, wherein when the calculated platelet separation index DI is 1/3 ≦ DI < 1, the calibrating comprises:
the measured platelet aggregation rate results are calibrated by a mapping function comprising a polynomial function, the calibration aimed at minimizing the error between the measured results and those measured when DI ≧ 1.
3. A calibration method of a platelet aggregation meter is characterized by comprising the following steps: the method for determining a result of measurement of platelet aggregation rate according to any one of claims 1 to 2;
and calibrating whether the measurement result of the platelet aggregation instrument is reliable or not according to the accuracy of the platelet aggregation rate determined by the platelet aggregation rate measurement result determination method.
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