CN115792198B - 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 32
- 230000002776 aggregation Effects 0.000 title claims abstract description 20
- 238000004220 aggregation Methods 0.000 title claims abstract description 20
- 208000013544 Platelet disease Diseases 0.000 title claims abstract description 18
- 208000010110 spontaneous platelet aggregation Diseases 0.000 claims abstract description 42
- 210000004369 blood Anatomy 0.000 claims abstract description 28
- 239000008280 blood Substances 0.000 claims abstract description 28
- 230000005856 abnormality Effects 0.000 claims abstract description 4
- 238000004519 manufacturing process Methods 0.000 claims abstract description 3
- 230000035945 sensitivity Effects 0.000 claims description 6
- 208000001490 Dengue Diseases 0.000 claims description 4
- 206010012310 Dengue fever Diseases 0.000 claims description 4
- 208000025729 dengue disease Diseases 0.000 claims description 4
- 238000012360 testing method Methods 0.000 abstract description 7
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 6
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- 210000001772 blood platelet Anatomy 0.000 description 37
- 238000001514 detection method Methods 0.000 description 8
- 239000003146 anticoagulant agent Substances 0.000 description 4
- 229940127219 anticoagulant drug Drugs 0.000 description 4
- 238000003745 diagnosis Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- 230000002159 abnormal effect Effects 0.000 description 2
- 230000008859 change Effects 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
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000013642 negative control 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
- 239000000243 solution Substances 0.000 description 2
- 208000007536 Thrombosis Diseases 0.000 description 1
- 230000004931 aggregating effect Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 210000003989 endothelium vascular Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000002947 procoagulating effect Effects 0.000 description 1
- 230000003578 releasing effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000009666 routine test Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
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- 239000002699 waste material Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/49—Blood
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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 platelet counting; s2, alarming abnormality of a blood cell analyzer, and reading each increment value b of the platelet count of the analyzer; s3, taking an alarm sample to manufacture a blood smear and performing microscopic examination, and reading a microscopic examination platelet aggregation value a and a field number n under a microscopic oil mirror field; s4, calculating a 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 value of c is less than or equal to the threshold value, judging that the sample is negative. Compared with the prior art, the invention has the remarkable advantages that: 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 beneficial 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 an important component of blood cells, have the functions of adhering, aggregating, releasing, procoagulating and contracting blood clots, and can maintain the integrity of vascular endothelium, thereby achieving the effect of stopping blood coagulation. The number of platelets directly affects the anticoagulant function of the patient. Platelet count is the basis of platelet parameter reliability, and accuracy of detection results is crucial. Platelet count accuracy is affected by many factors, platelet aggregation is one of the more common interfering factors, leading to a pseudo-decrease in platelets (PTCP). PTCP may occur in healthy or diseased individuals of different ages and sexes. Existing automatic blood cell analyzers tend to have higher sensitivity indicators, but relatively lower specificity indicators.
Document 1: meng Lingzhang the sensitivity of the Sysmex NX-1000 blood cell analyzer platelet aggregation alarm was 98.15% and the specificity was 63.5% for the platelet aggregation alarm, medical research and education, volume 38, 2, month 4, 2021.
When the automatic blood cell analyzer prompts abnormal platelet aggregation alarm, an inspector often contacts a patient to re-collect blood by using different anticoagulant blood collection tubes so as to correct the influence of platelet aggregation on platelet count. However, since the existing blood cell analyzer has relatively low specificity, the detection result is negative after the blood is collected again. Therefore, the waste of the consumable of the test reagent is caused, and the time for the patient to acquire the report and seek medical attention is greatly delayed.
Therefore, there is a need in the clinic to establish a set of methods for assisting in determining abnormal platelet aggregation by an automated blood cell analyzer.
Disclosure of Invention
The invention aims to provide a method and a system for assisting a blood cell analyzer in judging abnormal platelet aggregation, so as to solve the problem of low specific index of the existing automatic blood cell analyzer.
The technical solution for achieving the purpose of the invention is as follows:
a method of assisting a blood cell analyzer in determining abnormal platelet aggregation, comprising the steps of:
s1, taking a blood sample, and placing the blood sample into a blood cell analyzer for platelet counting;
s2, alarming abnormality of a blood cell analyzer, and reading each increment value b of the platelet count of the analyzer;
s3, taking an alarm sample to manufacture a blood smear and performing microscopic examination, and reading a microscopic examination platelet aggregation value a and a field number n under a microscopic oil mirror field;
s4, calculating a 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 value of c is less than or equal to the threshold value, judging that the sample is negative.
Further, the c threshold is obtained by the following method: the area under the working curve of the subject is obtained through the working characteristic curve of the platelet aggregation index c, the maximum point of the about dengue index is found through calculating the about dengue index= (sensitivity + specificity) -1, and the threshold value of c is calculated according to the maximum point. Preferably, the threshold value of the platelet aggregation index c is 21.36.
Further, the number n of the fields of view under the microscope oil mirror field is more than or equal to 100.
Further, the blood cell analyzer is an electrical impedance method blood cell analyzer.
Correspondingly, the invention also provides a system for judging abnormal platelet aggregation by using the auxiliary blood cell analyzer, 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 an instrument platelet count value a; the microscopic examination module is used for microscopic examination of the platelet abnormal aggregation alarming sample, and reading a microscopic platelet aggregation value a and a field number n of a microscope under the field of view; 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 remarkable advantages that: 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 beneficial to the current platelet counting method.
Drawings
FIG. 1 is a schematic flow chart of the working method of the invention.
FIG. 2 is a graph of platelet count versus platelet aggregation for the present invention.
FIG. 3 is a graph showing the comparison of the platelet aggregation index c after correction according to the present invention in the positive and negative control groups.
Fig. 4 is a graph of a diagnosis of abnormal platelet aggregation.
Fig. 5 is a platelet reporting alert intent of a hematology analyzer:
wherein FIG. 5A is a platelet reporting alert intent of Sysmex XN; fig. 5B is a platelet reporting intent of BC 7500.
Detailed Description
The preferred embodiments of the present invention will be described below with reference to the accompanying drawings, it being understood that the preferred embodiments described herein are for illustration and explanation of the present invention only, and are not intended to limit the present invention.
1. Object(s)
Example 1 blood routine test platelet aggregation alarm samples from first hospital hospitalization in south kyo city were collected 55 cases within 190 days of the first random choice from 2022, 2 months to 2022, 7 months.
Example 2 platelet aggregation alarm samples were collected 20 cases for clinical Sysmex XN and Michael BC7500 blood cell analyzer of patients hospitalized in first hospital in Nanj, city at the beginning of month 2022.
Sample basic information and related blood routine data are collected, including patient age, sex, diagnosis, blood routine detection platelet count value, and re-collection of corrected platelet count value.
2. Instrument and reagent
The method comprises the steps of detecting by using a Michaelis blood cell analyzer BC-7500 or a Heiden blood cell analyzer XN-2800 full-automatic blood cell analyzer, and using reagents and calibrator matched with the instrument.
Microscope model OLMPUS CX21, company of olympus corporation, japan
BDEDTA-K2 vacuum anticoagulation tube in U.S.
Jimssa stain (Pearl sea Soy Biotechnology Co., china)).
3. Detection method
(1) Platelet detection after the LIS system number is checked by an operation program hospital on an instrument, the platelet detection is directly put into a test tube rack and then is detected by a blood cell analyzer BC-7500 of Michael company or a blood cell analyzer XN-2800 of Hessenkel company. As shown in fig. 5, when platelet aggregation is detected in the blood, the instrument automatically alarms for platelet aggregation (PLT Clumps).
(2) Step study of blood smears and swiss staining follow the national clinical test protocol (fourth edition) with reference to manual microscopy, the specific steps are as follows:
(1) thoroughly mixing suspected platelet aggregation specimen, and adding a drop of blood sample (about 40-50 ul) at 1/3 of the proximal end of the slide;
(2) the blood drop is rapidly dispersed along the push piece at an angle of 30-45 degrees, and the push piece is rapidly and stably pushed to the other end of the glass slide;
(3) after the blood sheet is dried, the following steps are carried out according to 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 dye liquor is washed away by running water, and the platelet aggregation number is counted under a microscope after the dye liquor is dried.
4. Positive and negative judgment criteria:
as shown in fig. 5, the platelet aggregation or abnormal platelet histogram alarm sample is re-detected by the same instrument after blood collection by using other anticoagulants (such as sodium citrate) blood collection tubes, and the significant change (> 20%) of the platelet count is determined to be a normal platelet abnormal aggregation sample, positive. Otherwise, if the platelet count and the alarm clock value after blood collection are less than or equal to 20%, the normal physiological aggregation count sample of the platelets is judged to be negative.
Example 1: formula reasoning
Manual microscopic examination was performed on a normal physiological aggregate count sample of 20 blood platelets. When manual microscopic examination is performed, at least 100 visual fields are counted according to the sequence from left to right and from top to bottom, so that the accuracy and reliability of results are ensured. Otherwise, the view field is insufficient, and the platelet aggregation number may not be accurately counted, resulting in deviation of the result. Meanwhile, in the manual microscopic examination, the platelet aggregation number a under the field of n oil mirrors (objective lens multiplied by 100) is uniformly corrected to be the platelet aggregation number n' under 1000 fields. And carrying out 20 sample position marks by taking the microscopic platelet aggregation value a as an ordinate and taking each increment value b of the platelet count of the instrument as an abscissa, and finally obtaining the result of the figure 2, wherein a and b are positively correlated.
According to the existing standard, the value b of the platelet count per increment of the instrument is less than 100X 10 9 Is often used as a low platelet sample and is proposed to clinically bleed windRisk indicator (less than 100X 10) 9 L is a critical value for clinical intervention or diagnosis of a patient), so that the positive and negative samples of each increment b of the platelet count of the instrument are uniformly corrected to be 100X 10 in platelet count 9 The platelet aggregation index c, the final formula is as follows:
( n is the view count (n is more than or equal to 100) under the microscope oil-scope view (object lens multiplied by 100); platelet aggregation number; and b, counting platelets of the instrument. )
The platelet aggregation index c required for final statistical verification is calculated according to the above formula, and by comparing the positive sample with the negative control, the positive and negative platelet aggregation indexes c are found to be significantly increased as shown in fig. 3 (p < 0.05).
The platelet aggregation index c subject work characteristic curve (receiver operating characteristic curve, ROC) was plotted with 29 cases (including the above 20 normal physiological platelet aggregation count samples) as a negative group and 26 positive samples, and as shown in fig. 4, the c value had a statistical significance, and the area under the subject work curve (area under the receiver operating characteristic curve, AUC) was obtained, and the critical (cut-off) value was calculated by calculating the jouden index (you den' sindex, YI), yi= (sensitivity + specificity) -1, finding the YI maximum point, and calculating the critical (cut-off) value from 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 platelet aggregation index c threshold, the sensitivity was 92.3% and the specificity was 86.2%.
Example 2: method implementation and effect verification
The effect verification of the invention is carried out by using a specimen with platelet alarm of an electrical impedance method blood cell analyzer. The electrical impedance method blood cell analyzer used in this example was a Sysmex XN and a Michael BC7500 blood cell analyzer. The 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 platelet counting;
s2, alarming abnormality of a blood cell analyzer, and reading each increment value b of the platelet count of the analyzer;
s3, taking an alarm sample, making a blood smear, performing microscopic examination, and reading a microscopic examination platelet aggregation value a and the number n of fields under the microscopic oil microscopic field (at least 100 fields are counted in the order from left to right and from top to bottom during microscopic examination);
s4, calculating a 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 value of c is less than or equal to the threshold value, judging that the sample is negative.
The specific results are as follows:
meanwhile, the 20 cases of alarm samples are detected by adopting a prior art method, sodium citrate is taken as an anticoagulant, blood of a patient is collected again, the change condition of the platelet count of the patient is analyzed by using a Sysmex XN or a MiceBC 7500 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 steps:
taking sample number 1,3,5,9,12 as an example, the number of platelets obtained in the first blood cell analyzer was all lower than 100×10 9 L belongs to clinical intervention values, and the result obtained by the method is consistent with the existing clinical re-determination standard.
Taking sample number 17 as an example, the number of platelets obtained in the first blood cell analyzer is higher than300×10 9 L is a clinical intervention value, and the results obtained by the method are consistent with the existing clinical re-determination standard.
This means that the method provided by the invention is not only suitable for (100-300) x 10 with clinical count value in the normal range 9 the/L instrument alarm detection is also suitable for instrument alarm detection of clinical numerical values needing intervention.
The method for judging abnormal platelet aggregation by the auxiliary blood cell analyzer provided by the invention can judge the alarm of the existing blood cell analyzer, has a wide trial range, is beneficial to supplement to the existing reinspection rule, saves reagent cost, relieves the pain of patients and improves the laboratory test efficiency.
It will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention also include such modifications and alterations insofar as they come within the scope of the appended claims or the equivalents thereof.
Claims (1)
1. A method of assisting a hematology 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 platelet counting;
s2, alarming abnormality of a blood cell analyzer, and reading each increment value b of the platelet count of the analyzer;
s3, taking an alarm sample to manufacture a blood smear, performing microscopic examination, and reading a microscopic examination platelet aggregation value a
And a field number n under the microscope oil field;
s4, calculating a 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; when the value c is less than or equal to the threshold value, judging that the sample is negative;
platelet aggregation index c the subject working characteristic curve obtains the area under the subject working curve, the maximum point of the about dengue index is found by calculating the about dengue index= (sensitivity + specificity) -1, and the threshold value of c is calculated by the point;
the threshold value of the platelet aggregation index c was 21.36;
the number n of the fields of view of the microscope oil mirror is more than or equal to 100;
the blood cell analyzer in the step S1 is an electrical impedance method blood cell analyzer;
the electrical impedance method blood cell analyzer 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 an instrument platelet count value a;
the microscopic examination module is used for microscopic examination of a sample with abnormal platelet aggregation alarm and reading a microscopic platelet aggregation value a and a field number n of a microscope under the field of view;
the threshold judgment module is used for calculating the platelet aggregation index c and judging the threshold.
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| PCT/CN2022/118419 WO2024050856A1 (en) | 2022-09-06 | 2022-09-13 | Method and system for assisting blood cell analyzer in determining abnormal platelet aggregation |
| 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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