CN115792198A - Method and system for assisting blood cell analyzer in judging abnormal platelet aggregation - Google Patents
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- 210000000601 blood cell Anatomy 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 35
- 230000002776 aggregation Effects 0.000 title claims abstract description 24
- 238000004220 aggregation Methods 0.000 title claims abstract description 24
- 208000013544 Platelet disease Diseases 0.000 title claims abstract description 20
- 210000001772 blood platelet Anatomy 0.000 claims abstract description 47
- 208000010110 spontaneous platelet aggregation Diseases 0.000 claims abstract description 41
- 210000004369 blood Anatomy 0.000 claims abstract description 28
- 239000008280 blood Substances 0.000 claims abstract description 28
- 230000000007 visual effect Effects 0.000 claims abstract description 20
- 230000035945 sensitivity Effects 0.000 claims description 6
- 238000000386 microscopy Methods 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 10
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 6
- 230000002159 abnormal effect Effects 0.000 abstract description 4
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- 239000013589 supplement Substances 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract 1
- 238000003745 diagnosis Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
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- 230000000694 effects Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 2
- RKQKLZMMOQWTGB-HYBUGGRVSA-N diphenyl-[(1R,2S)-2-(phenylsulfanylmethyl)cyclopentyl]phosphane Chemical compound C([C@@H]1[C@@H](CCC1)P(C=1C=CC=CC=1)C=1C=CC=CC=1)SC1=CC=CC=C1 RKQKLZMMOQWTGB-HYBUGGRVSA-N 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 239000003805 procoagulant Substances 0.000 description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
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Abstract
The invention relates to a method and a system for assisting a blood cell analyzer in judging abnormal platelet aggregation. The method comprises the following steps: s1, taking a blood sample, and placing the blood sample into a blood cell analyzer for counting blood platelets; s2, alarming by the blood cell analyzer in an abnormal way, and reading the value b of each liter of platelet count of the analyzer; s3, taking an alarm sample, making a blood smear, performing microscopic examination, and reading a microscopic platelet aggregation value a and a visual field number n under a microscopic oil microscope visual field; s4, calculating the platelet aggregation index c according to the following formula:
s5, setting a threshold value of c, and judging that the sample is positive when c is larger than the threshold value; and when the c value is less than or equal to the threshold value, judging the sample to be negative. Compared with the prior art, the invention has the following remarkable advantages: by calculating the platelet aggregation index c, abnormal platelets of the blood cell analyzer can be remarkably reducedThe error rate of alarm is gathered, the material consumption of the detection reagent is saved, the detection efficiency is improved, and the method is a beneficial supplement to the current platelet counting method.
Description
Technical Field
The invention relates to the technical field of in-vitro diagnosis, in particular to a method and a system for assisting a blood cell analyzer in judging abnormal platelet aggregation.
Background
Platelets are important components of blood cells, have the functions of adhesion, aggregation, release, procoagulant and clot contraction, and can maintain the integrity of vascular endothelium, thereby achieving the effect of stopping blood coagulation. The number of platelets directly affects the procoagulant function of the patient. The platelet count is the basis of the reliability of platelet parameters, and the accuracy of the detection result is very important. Platelet count accuracy is influenced by a number of factors, and platelet aggregation is one of the more common interfering factors, leading to false reduction of platelets (PTCP). PTCP can occur in healthy or diseased individuals of different ages and sexes. The existing automatic blood cell analyzer is high in sensitivity index, but low in specificity index.
Document 1: sysmex NX-1000 blood cell analyzer platelet aggregation alarm information analysis, medical research and education, no. 38, no. 2, no. 2021, 4.15% of platelet aggregation alarm sensitivity, 63.5% of specificity.
When the automatic blood cell analyzer prompts an alarm of abnormal platelet aggregation, the inspector often contacts the patient to take blood again using different anticoagulant blood collection tubes to correct the influence of platelet aggregation on platelet counting. However, the existing blood cell analyzer has low specificity, so the detection result is negative after blood is collected again. Thereby causing the waste of the test reagent consumables and greatly delaying the time for patients to obtain reports and seek medical advice in time.
Therefore, there is a need to establish a set of automatic blood cell analyzer for assisting in determining abnormal platelet aggregation.
Disclosure of Invention
The invention aims to provide a method and a system for assisting a blood cell analyzer in judging abnormal platelet aggregation, which aim to solve the problem of low specificity index of the existing automatic blood cell analyzer.
The technical solution for realizing the purpose of the invention is as follows:
a method for assisting a blood cell analyzer in judging abnormal platelet aggregation comprises the following steps:
s1, taking a blood sample, and placing the blood sample into a blood cell analyzer for counting blood platelets;
s2, alarming abnormally by the blood cell analyzer, and reading a value b of each liter of platelet count of the analyzer;
s3, taking an alarm sample, making a blood smear, performing microscopic examination, and reading a microscopic platelet aggregation value a and a visual field number n under a microscopic oil microscope visual field;
s4, calculating the platelet aggregation index c according to the following formula:
s5, setting a threshold value of c, and judging that the sample is positive when c is larger than the threshold value; and when the c value is less than or equal to the threshold value, judging the sample to be negative.
Further, the c threshold is obtained by the following method: and (3) obtaining the area under the working curve of the subject according to the platelet aggregation index c and the working characteristic curve of the subject, and calculating the point of the maximum Johnson index by calculating the Johnson index = (sensitivity + specificity) -1, so as to calculate the threshold value of c. Preferably, the threshold value for the platelet aggregation index c is 21.36.
Furthermore, the number n of visual fields under the oil microscope visual field is more than or equal to 100.
Furthermore, the blood cell analyzer is an electrical impedance method blood cell analyzer.
Correspondingly, the invention also provides a system for applying the method for assisting the blood cell analyzer in judging the abnormal platelet aggregation, which comprises a blood cell analyzer module, a microscopic examination module and a threshold judgment module; the blood cell analyzer module is used for alarming abnormal platelet aggregation and reading a platelet count value a of the analyzer; the microscopic examination module is used for performing microscopic examination on the sample for alarming abnormal platelet aggregation and reading a microscopic platelet aggregation value a and a visual field number n under a microscopic oil microscope visual field; the threshold judgment module is used for calculating the platelet aggregation index c and judging the threshold.
Compared with the prior art, the invention has the following remarkable advantages: by calculating the platelet aggregation index c, the error rate of abnormal platelet aggregation alarm of the blood cell analyzer can be obviously reduced, the consumption of test reagents is saved, the test efficiency is improved, and the method is a beneficial supplement to the current platelet counting method.
Drawings
FIG. 1 is a schematic flow chart of the working method of the present invention.
FIG. 2 is a graph of the relationship between platelet count and platelet aggregation according to the present invention.
FIG. 3 is a graph comparing the corrected platelet aggregation index c of the present invention in positive and negative controls.
Fig. 4 is a graph showing a diagnosis of abnormal platelet aggregation.
FIG. 5 is a schematic of a platelet alarm for a blood cell analyzer:
wherein fig. 5A is a schematic platelet alarm diagram of Sysmex XN; fig. 5B is a schematic diagram of a platelet alarm for BC 7500.
Detailed Description
The preferred embodiments of the present invention will be described in conjunction with the accompanying drawings, and it will be understood that they are described herein for the purpose of illustration and explanation and not limitation.
1. Object
Example 1 platelet aggregation alarm samples 55 were routinely tested to collect 55 platelet aggregation alarm samples from inpatients at the first hospital in Nanjing City within 190 days randomly selected from 2 months at 2022 to 7 months at 2022.
Example 2 to collect 20 clinical Sysmex XN and mreiy BC7500 platelet aggregation alarm samples of inpatients at first hospital in Nanjing City at 8 months of 2022.
Sample basic information and associated blood routine data including patient age, gender, diagnosis, blood routine test platelet count values and re-collection of corrected platelet count values are collected.
2. Instruments and reagents
The method adopts a Mry company blood cell analyzer BC-7500 or a Hesimecon company blood cell analyzer XN-2800 full-automatic blood cell analyzer for detection, and uses a reagent and a calibrator matched with the analyzer.
Microscope model OLMPUS CX21, olympus corporation, japan
U.S. BDEDTA-K2 vacuum anticoagulation tube.
Giemsa rui (sekkiso biotechnology, sekkiso, china)).
3. Detection method
(1) The platelet detection is carried out on an instrument, after the serial number of an LIS system is detected by a hospital according to an operation program, the platelet is directly placed into a test tube rack and then is detected by a blood cell analyzer BC-7500 of Mirui company or a blood cell analyzer XN-2800 of Hesimecon company. As shown in fig. 5, the instrument automatically alarms for platelet aggregation (PLT platelets) when the detection reveals platelet aggregation in the blood.
(2) The step study of blood smear and Swiss staining follows the national operating rules of clinical examination (fourth edition) with reference to artificial microscopic examination, and comprises the following specific steps:
(1) mixing the suspected platelet aggregation specimen, adding a drop of blood sample (about 40-50 ul) at 1/3 of the near end of the slide;
(2) the blood drops are quickly dispersed along the push sheet at an angle of 30-45 degrees, and the push sheet is quickly and stably pushed to the other end of the glass slide;
(3) after the blood slices are dried, the method comprises the following steps of 1:1 adding a plurality of drops of Swiss dye solution and PH6.8 phosphate buffer solution to cover the whole blood membrane;
(4) after 5 minutes, the staining solution is flushed by running water, and the number of platelet aggregates is counted under a microscope after the staining solution is dried.
4. Positive and negative judgment criteria:
as shown in fig. 5, the platelet aggregation or platelet histogram abnormal alarm specimen is re-detected by the same instrument after blood collection by using other anticoagulant (such as sodium citrate) blood collection tubes, and the platelet count is significantly changed (> 20%) and is determined to be positive in the routine platelet abnormal aggregation specimen. And otherwise, if the counting value of the blood platelets after blood collection and the counting value of the alarm timer are less than or equal to 20 percent, the normal physiological aggregation counting sample of the blood conventional blood platelets is judged to be negative.
Example 1: formula reasoning
20 samples of normal physiological platelet aggregation counts from normal platelets were manually microscopically examined. When counting is carried out during manual microscopic examination, at least 100 visual fields need to be counted in a city wall manner from left to right and from top to bottom so as to ensure the accuracy and reliability of results. Otherwise, the visual field is not enough, and the aggregation number of the platelets can not be accurately counted, so that the result is deviated. Meanwhile, in the manual microscopic examination, the platelet aggregation number a under n oil-immersed mirrors (objective lens × 100) visual fields is uniformly corrected to be the platelet aggregation number n' under 1000 visual fields. And marking the positions of 20 samples by taking the platelet aggregation value a of microscopic examination as a vertical coordinate and taking the platelet counting value b of the instrument as a horizontal coordinate, and finally obtaining the result shown in the figure 2, wherein a is positively correlated with b.
The platelet count of the instrument is less than 100X 10 per liter according to the existing standard 9 Is often used as a low platelet sample and as an index (less than 100X 10) for clinical bleeding risk 9 L is a critical value for clinical intervention or diagnosis of the patient), therefore the positive and negative samples per liter of b of the platelet count of the instrument are corrected to a platelet count of 100 x 10 9 Platelet aggregation index c, the final formula is as follows:
( n is the view count under the microscope oil-scope view (object lens multiplied by 100) (n is more than or equal to 100); a is platelet aggregation number; and b, counting platelets by using the instrument. )
The platelet aggregation index c required for the final statistical validation is calculated according to the formula, and the platelet aggregation index c which is positive and negative is found to be remarkably increased by comparing the positive sample with the negative control as shown in figure 3 (p < 0.05).
Using 29 cases (including 20 cases of the above-mentioned blood conventional platelet normal physiological aggregation count samples) as a negative group and 26 cases of positive samples to draw a platelet aggregation index c subject working characteristic curve (ROC), as shown in fig. 4, the c value has a statistical significance, the area under the subject working curve (AUC) is obtained, and by calculating the approximate index (Youden's index, YI), YI = (sensitivity + specificity) -1, the YI maximum point is found, and the cut-off value is calculated at this point. Through calculation, the optimal cut off value is 21.36, and the area under the curve is 99.4%. As shown in fig. 4, when 21.36 was used as the threshold of the platelet aggregation index cghreshold, the sensitivity was 92.3% and the specificity was 86.2%.
Example 2: method implementation and effect verification
The effect of the invention is verified by using a sample for platelet alarm of an electrical impedance blood cell analyzer. The electrical impedance method blood cell analyzer used in this example was a Sysmex XN and Merrill BC7500 blood cell analyzer. Other manufacturers and models of blood cell analyzers can be judged by the method.
The method steps shown in fig. 1 are as follows:
s1, taking a blood sample, and placing the blood sample into a blood cell analyzer for counting blood platelets;
s2, alarming by the blood cell analyzer in an abnormal way, and reading the value b of each liter of platelet count of the analyzer;
s3, taking the alarm sample to prepare a blood smear and performing microscopic examination, and reading a microscopic platelet aggregation value a and a visual field number n under a microscopic oil-scope visual field (at least 100 visual fields are counted in a city wall manner according to the sequence from left to right and from top to bottom during microscopic examination);
s4, calculating the platelet aggregation index c according to the following formula:
s5, setting a threshold value (21.36) of c, and judging that the sample is positive when c is greater than the threshold value; and when the c value is less than or equal to the threshold value, judging the sample to be negative.
The specific results are as follows:
and simultaneously, the 20 alarm samples are detected by adopting the prior art method, sodium citrate is taken as anticoagulant, the blood of the patient is collected again, the platelet count change condition of the patient is analyzed by a Sysmex XN or Merrill BC7500 full-automatic blood cell analyzer with the same model, and finally the experimental result is consistent with the result of the judging method adopted by the invention. The method comprises the following specific steps:
for example, sample numbers 1,3,5,9,12, platelet numbers obtained in the first time hematology analyzer were all below 100X 10 9 the/L belongs to clinical intervention values, and the results obtained by the method adopted by the invention are consistent with the existing clinical re-determination standards.
Taking sample number 17 as an example, the number of platelets obtained in the primary blood cell analyzer is higher than 300 × 10 9 the/L belongs to clinical intervention values, and the results obtained by the method adopted by the invention are consistent with the existing clinical re-determination standards.
This demonstrates that the method provided by the present invention is not only applicable to (100-300). Times.10 clinical counts in the normal range 9 the/L instrument alarm detection is also suitable for the instrument alarm detection of which clinical values need to intervene.
The method for assisting the blood cell analyzer in judging abnormal platelet aggregation can be used for judging the alarm of the conventional blood cell analyzer, has a wide trial range, is beneficial to supplement the conventional reinspection rule, saves the reagent cost, relieves the pain of patients and improves the laboratory inspection efficiency.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (6)
1. A method of assisting a blood cell analyzer in determining abnormal platelet aggregation, the method comprising the steps of:
s1, taking a blood sample, and placing the blood sample into a blood cell analyzer for counting blood platelets;
s2, alarming abnormally by the blood cell analyzer, and reading a value b of each liter of platelet count of the analyzer;
s3, taking an alarm sample, making a blood smear, performing microscopic examination, and reading a microscopic platelet aggregation value a and a visual field number n under a microscopic oil microscope visual field;
s4, calculating the platelet aggregation index c according to the following formula:
s5, setting a threshold value of c, and judging that the sample is positive when c is larger than the threshold value; and when the c value is less than or equal to the threshold value, judging the sample to be negative.
2. The method of claim 1, wherein the platelet aggregation index c is an area under a subject operation curve obtained from a subject operation characteristic curve, and the threshold value c is calculated by calculating a jotans index = (sensitivity + specificity) -1, and finding a point of maximum jotans index, based on the point.
3. The method of claim 2, wherein the threshold value of the platelet aggregation index c is 21.36.
4. The method for determining abnormal platelet aggregation according to claim 1, wherein the number of visual fields n in the oil-scope field of the microscope is not less than 100.
5. The method as claimed in claim 1, wherein the blood cell analyzer in step S1 is an electrical impedance blood cell analyzer.
6. A system for applying the method of assisting a blood cell analyzer in determining abnormal platelet aggregation according to any one of the preceding claims, comprising a blood cell analyzer module, a microscopy module and a threshold determination module;
the blood cell analyzer module is used for alarming abnormal platelet aggregation and reading a platelet count value a of the analyzer;
the microscopic examination module is used for performing microscopic examination on the sample for alarming abnormal platelet aggregation and reading a microscopic platelet aggregation value a and a visual field number n under a microscope oil microscope visual field;
the threshold judgment module is used for calculating the platelet aggregation index c and judging the threshold.
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| CN202211083734.1A CN115792198B (en) | 2022-12-01 | 2022-12-01 | Method and system for assisting blood cell analyzer in judging abnormal platelet aggregation |
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