WO2019015611A1 - 提高白细胞分类结果准确性和计数结果重复性的方法及设备 - Google Patents

提高白细胞分类结果准确性和计数结果重复性的方法及设备 Download PDF

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WO2019015611A1
WO2019015611A1 PCT/CN2018/096143 CN2018096143W WO2019015611A1 WO 2019015611 A1 WO2019015611 A1 WO 2019015611A1 CN 2018096143 W CN2018096143 W CN 2018096143W WO 2019015611 A1 WO2019015611 A1 WO 2019015611A1
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blood cell
white blood
sample
added
counting
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习武佳
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深圳市帝迈生物技术有限公司
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/00584—Control arrangements for automatic analysers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1009—Characterised by arrangements for controlling the aspiration or dispense of liquids

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  • the present invention relates to the field of blood analyzers, and more particularly to a method and apparatus for improving the accuracy of white blood cell classification results and the repeatability of counting results.
  • the three-class animal blood cell analyzer which is sold on the market by the electrical impedance method is widely used in animal clinical and laboratory fields.
  • the accuracy and repeatability are evaluated by blood cell analysis.
  • An important indicator of instrument performance For the white blood cell count and classification of animal blood samples such as cats and dogs, the following three methods are mainly used:
  • the first method is to treat the blood sample of the animal according to the same hemolysis dose of the human blood sample. This method can ensure that the blood sample of cats, dogs and other animals does not affect the white blood cell count after the hemolytic agent, but it is easy to cause the white blood cell classification. The exact problem is that the lymphocyte test results are high.
  • the second method is to dissolve the blood samples of cats, dogs and other animals, that is, to reduce the hemolysis dose.
  • the red blood cells do not affect the white blood cell count after the hemolytic agent, and the classification can be improved by comparing with the microscopic examination classification data.
  • the accuracy of the results Due to the reduction of the amount of hemolytic agent added, a certain proportion of samples will be interfered with by red blood cell debris, resulting in inaccurate counting results and classification results.
  • the red blood cell fragments are filtered out by fixed landmarks, and the classification of dogs, cats, etc. is not classified.
  • the quasi-problem can improve white blood cell classification performance by this method.
  • the use of fixed landmarks results in a large proportion of white blood cell counts for a significant proportion of animal samples.
  • the use of fixed landmarks to filter out red blood cell debris interferes with inaccurate white blood cell count results.
  • the third method adds the hemolytic agent twice in the white blood cell counting process, and tests the first white blood cell count value and the first white blood cell histogram after the first addition of the hemolytic agent, and the second time.
  • the second white blood cell count value after the addition of the hemolytic agent; the first added hemolysis dose, after the action, the blood sample still has a certain amount of red blood cell debris affecting the white blood cell count value, and the second added hemolysis dose, after the action There is no effect of red blood cell debris in the white blood cell count value; according to the ratio of the first white blood cell count value to the second white blood cell count value, the histogram of white blood cell classification is determined, and white blood cell classification is performed, which ensures the accuracy of the white blood cell count value and the hemoglobin test result.
  • Sex but also improve the accuracy of white blood cell classification results in blood samples of dogs, cats and other animals.
  • This method uses a split blood, and separately adds hemolytic agent to white blood cell count twice. Finally, the ratio of the first count and the second count result is used as a judgment basis, and the white blood cell count and classification result are output, although most samples can be guaranteed. The accuracy of the classification and counting results, but the reproducibility of the blood cytometer remains to be further improved.
  • the object of the present invention is to provide a method and a device for improving the accuracy of white blood cell classification results and the repeatability of counting results, aiming at solving the problem that the detection results of the existing three-class animal blood cell analyzer are inaccurate and the repeatability is poor.
  • An apparatus for improving the accuracy of white blood cell classification results and repeatability of counting results comprising: a white blood cell counting device for white blood cell counting;
  • a sample blood pushing device for dispensing a blood sample reagent to a white blood cell counting device
  • a hemolytic agent pushing device for dispensing a hemolysis reagent to a white blood cell counting device
  • An automatic control system for separately controlling the sample blood pushing device and the hemolytic agent pushing device to push the sample blood and the hemolytic agent to the white blood cell counting device in batches;
  • a cleaning and draining device for cleaning the white blood cell counting device
  • An accuracy comparison module for comparing the results of each classification count
  • the automatic control system controls the sample blood pushing device and the hemolytic agent pushing device to push the reagent to the white blood cell counting device twice, and the amount of the blood sample pushed for the first time is the same as the amount of the second blood sample, and the second hemolysis The dose is less than the first hemolysis dose.
  • a method for improving the accuracy of white blood cell classification results and the repeatability of counting results comprising the following steps:
  • Step S1 first dividing the blood to the white blood cell counting device and adding the first hemolytic agent, and the first hemolysis dose is recorded as L1;
  • Step S2 the white blood cell technology device performs the first white blood cell count, and is recorded as WBC1;
  • Step S3 emptying and cleaning the white blood cell counting device
  • Step S4 dividing the blood to the white blood cell counting device for the second time and adding a second hemolytic agent, the second hemolytic dose is recorded as L2, and controlling L1 is greater than L2;
  • Step S5 the white blood cell technology device performs a second white blood cell count, which is recorded as WBC2;
  • Step S6 taking the ratio of the first counting result WBC1 and the second result WBC2 as a judgment basis, confirming the area of the red blood cell fragment area by the area ratio, and the total area removing the red blood cell fragment area area is the second white blood cell counting classification result.
  • Step S7 The first count and the second count of the number of particles are added together to statistically output the white blood cell count result.
  • the ratio of the first counting result WBC1 to the second result WBC2 is used as a basis for judging, and the specific method for confirming the area of the red blood cell fragment area by the area ratio is:
  • WBC2/WBC1 (Sr+Sw)/Sw
  • the value of the second hemolysis dose L2 is determined by the following steps:
  • Step a1 taking the first hemolysis dose L1 value as a reference, and testing the WBC1i with 50 samples;
  • Step a2 taking a plurality of values of 40%-70% of the above L1 value and evaluating 50 samples of the same standard, first observing the classification result of the white blood cells, and if the value of the white blood cell three classification cannot be achieved, the elimination is possible; The value of the apparent classification effect is used as the candidate value of L2.
  • the 50 sets of the same standard are respectively counted for the multiple sets of L2 alternative values, and the WBC2i of each candidate value is obtained, and the ratio of WBC2i and WBC1i is distributed between 1 and 1.6.
  • the most alternative value is used as the value of the second blood dose L2;
  • i is rounded from 1 to 50.
  • the invention has the beneficial effects that the invention determines the white blood cell count result and the classification result by two methods of dividing blood and adding hemolytic agent into the WBC pool (white blood cell counting pool), and the second counting ensures the accuracy of the WBC classification result. Then, the result of the first white blood cell count and the result of the second white blood cell count are added together, and the number of two-stage particles is counted to obtain the counting result of the white blood cells, the number of statistical particles of the sample is increased, and the WBC counting result of the blood cell analyzer is improved. Repeatability.
  • the program not only ensures the accuracy of the white blood cell classification results of the blood cell analyzer, but also improves the repeatability of the white blood cell count results.
  • FIG. 1 is a block diagram of a device module provided by the present invention.
  • FIG. 2 is a schematic flow chart of the method provided by the present invention.
  • Fig. 3 is a schematic diagram showing the histogram of the first white blood cell count in the present invention.
  • Fig. 4 is a schematic diagram showing the histogram of the second white blood cell count result in the present invention.
  • Fig. 5 is a view showing the area of the red blood cell fragment area and the area of white blood cells in the histogram of the second white blood cell count result in the present invention.
  • Fig. 6 is a schematic diagram showing a histogram of white blood cell counts after treatment in the present invention.
  • the object of the present invention is to provide a method for improving the accuracy of white blood cell classification results of blood cell analyzers and the repeatability of counting results, which not only ensures the accuracy of white blood cell classification results of blood cell analyzers, but also improves the repeatability of white blood cell counting results.
  • the main modules include:
  • a white blood cell counting device for white blood cell counting
  • a sample blood pushing device for dispensing a blood sample reagent to a white blood cell counting device
  • a hemolytic agent pushing device for dispensing a hemolysis reagent to a white blood cell counting device
  • An automatic control system for separately controlling the sample blood pushing device and the hemolytic agent pushing device to push the sample blood and the hemolytic agent to the white blood cell counting device in batches;
  • a cleaning and draining device for cleaning the white blood cell counting device
  • Accuracy comparison module for comparing the accuracy of each classification result.
  • the automatic control system controls the sample blood pushing device and the hemolytic agent pushing device to push the reagent to the white blood cell counting device twice.
  • the amount of blood sample pushed for the first time is the same as the amount of blood sample for the second time.
  • the first hemolysis dose is sufficient, and the second hemolysis dose is less than the first hemolysis dose.
  • the method flow provided by the present invention is as shown in FIG. 2, and includes the following steps:
  • Step S1 first dividing the blood to the white blood cell counting device and adding the first hemolytic agent, and the first hemolysis dose is recorded as L1;
  • Step S2 the white blood cell technology device performs the first white blood cell count, and is recorded as WBC1;
  • Step S3 emptying and cleaning the white blood cell counting device
  • Step S4 dividing the blood to the white blood cell counting device for the second time and adding a second hemolytic agent, the second hemolytic dose is recorded as L2, and controlling L1 is greater than L2;
  • Step S5 the white blood cell technology device performs a second white blood cell count, which is recorded as WBC2;
  • Step S6 using the ratio of the first counting result to the second result as the judgment basis, confirming the area of the red blood cell fragment area by the area ratio, that is, the first landmark value, and the total area removing the red blood cell fragment area is the second white blood cell. Counting the classification results;
  • Step S7 The first count and the second count of the number of particles are statistically outputted to the white blood cell count result.
  • the first WBC count is the result of counting the completely dissolved red blood cells, which can be directly output as the WBC count result.
  • the white blood cell particle count formula :
  • White blood cell count results total number of white blood cell effective particles / (dilution ratio * acquisition time)
  • the present invention adds the total number of effective particles of the first counting WBC1 and the second counting WBC2 (after considering the red blood cell debris), thereby outputting the counting result of the white blood cells, aiming at ensuring the accuracy of the result and the systematic deviation. Under the statistic increase, it is equivalent to reducing the influence of system deviation on the sample test results, and can improve the repeatability of white blood cell count results.
  • the first added hemolysis dose L1 is sufficient to ensure that more than 98% of the first blood samples of the animal are not interfered with by the red blood cell debris, and the first white blood cell count result is WBC1.
  • the lower white blood cell histogram as shown in Fig. 3, the value of the hemolysis dose L1 can be appropriately adjusted according to the dilution ratio. Among them, the sample blood sample input amount at each test is preset, and the same every time.
  • the selection of the first hemolysis dose L1 is also the optimal value obtained by a plurality of experimental adjustments in advance, and the hemolysis dose has been set in the detection application phase. Of course, the present invention does not exclude providing the first hemolysis dose.
  • L1 optional blood analyzer. The selectable range is also a prerequisite for ensuring that more than 98% of the animal blood samples added for the first time are white blood cell counts without interference with red blood cell debris.
  • the second hemolysis dose added to the white blood cell counting device is L2.
  • the second hemolysis dose L2 is smaller than the first hemolysis dose L1, that is, in the white blood cell counting device.
  • Animal blood samples were treated with weak solution, so that some red blood cell debris areas still exist, and the second white blood cell count result was WBC2, and a second white blood cell count histogram was obtained, as shown in Fig. 4.
  • L2 The value of L2 is determined during the experimental development stage. Each time we take the L2 optimal value (fixed value) that has been determined in the development stage, it can be input into the analyzer system for testing. It is not necessary to conduct an L2 value experiment for each test.
  • the specific value method is as follows:
  • the value of the second hemolysis dose L2 is determined by the following steps:
  • Step a1 taking the first hemolysis dose L1 value as a reference, and testing XBC samples to obtain WBC11, WBC12, WBC13...WBC1X;
  • Step a2 taking multiple values of 40%-70% of the above L1 value, and evaluating the X-sample of the same standard, first observing the classification result of white blood cells, and if the value of the three-class classification of white blood cells cannot be achieved, it can be eliminated; The value of the apparent classification effect is used as the candidate value of L2.
  • the multiple sets of L2 candidate values are counted by the same standard X-branch samples, and the WBC2i of each candidate value is obtained, and the ratio of WBC2i and WBC1i is distributed between 1 and 1.6.
  • the most alternative value is taken as the value of the second blood dose L2, i is taken to be 1 to X. Among them, X takes no less than 50.
  • WBC2/WBC1 (Sr+Sw)/Sw
  • Sr is the area where the red blood cell fragment area is counted for the second time
  • Sw is the area of the white blood cell counted for the second time
  • (Sr+Sw) is the area of the white blood cell histogram for the second time count.
  • the second counting result can output the classification result of the white blood cells, and the processed WBC histogram is as shown in FIG. 6. Compared with the actual microscopic examination results, the accuracy of white blood cell classification results is guaranteed.
  • Correction of the second white blood cell count result that is, after filtering out the interference of red blood cell debris, counting the number of white blood cells and the collection time of the first and second counts, the result of the first white blood cell count and the second white blood cell Counting the results of the number of particles added, the two-stage particle count to obtain the white blood cell count results, increase the sample statistical particle number, thereby improving the repeatability of the sample WBC count results.
  • the invention determines the white blood cell count result and the classification result by dividing the blood into the WBC pool twice and adding the hemolytic agent method, and the second counting ensures the accuracy of the WBC classification result. Then, the result of the first white blood cell count and the result of the second white blood cell count are added together, and the number of two-stage particles is counted to obtain the counting result of the white blood cells, the number of statistical particles of the sample is increased, and the WBC counting result of the blood cell analyzer is improved. Repeatability.

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Abstract

本发明公开了一种提高白细胞分类结果准确性和计数结果重复性的方法与设备，包括：白细胞计数的白细胞计数装置；向白细胞计数装置分配血样试剂的样本血推送装置；向白细胞计数装置分配溶血试剂的溶血剂推送装置；分别控制样本血推送装置和溶血剂推送装置分批次向白细胞计数装置推送样本血和溶血剂的自动控制系统；对白细胞计数装置进行清洗的清洗排液装置；对比每次分类计数结果的准确性的准确性对比模块；所述自动控制系统先后两次控制样本血推送装置和溶血剂推送装置向白细胞计数装置推送试剂，第二次溶血剂量小于第一次溶血剂量。采用本发明可提高了血液细胞分析仪WBC计数结果的准确性及重复性。

Description

提高白细胞分类结果准确性和计数结果重复性的方法及设备 【技术领域】
本发明涉及一种血液分析仪的领域，尤其涉及的是一种提高白细胞分类结果准确性和计数结果重复性的方法与设备。
【背景技术】
目前，市场上销售的以电阻抗法为检测原理的三分类动物血液细胞分析仪在动物临床及实验室领域应用广泛，对于三分类动物血液细胞分析仪，准确性以及重复性是评价血液细胞分析仪性能的重要指标。针对猫、狗等动物血液样本白细胞计数及分类，主要采用以下三种方法：
第一种方法是按照人血样本同样的溶血剂量对动物血样本进行处理，这种方法能够保证猫、狗等动物血液样本在溶血剂作用后红细胞碎片不影响白细胞计数，但容易造成白细胞分类不准确的问题，即淋巴细胞测试结果偏高。
第二种方法对猫、狗等动物血样本弱溶处理，即减少溶血剂量，对一定比例的样本，红细胞在溶血剂作用后不影响白细胞计数，通过与镜检分类数据的对比，可提高分类结果的准确性。由于减少溶血剂加入量，会造成一定比例的样本存在红细胞碎片干扰，导致计数结果和分类结果的不准确，目前通过固定界标，对红细胞碎片进行滤除，解决狗、猫等样本测试时分类不准的问题，通过这种方法可提高白细胞分类性能。但由于动物样本自身的特异性，用固定界标的方法，会导致有相当比例的动物样本的白细胞计数值有较大的偏差。用固定界标滤除红细胞碎片干扰后，会造成白细胞计数结果不准确。
第三种方法如专利CN 101470109 B，在白细胞计数过程中分先后两次加入溶血剂，并测试获得第一次加入溶血剂后的第一白细胞计数值和第一白细胞直方图，以及第二次加入溶血剂后的第二白细胞计数值；第一次加入的溶血剂量，其作用后使血液样本中仍有一定量影响白细胞计数值的红细胞碎片存在，第二次加入的溶血剂量，其作用后使白细胞计数值中没有红细胞碎片的影响；根据第一白细胞计数值和第二白细胞计数值之比，确定进行白细胞分类的直方图，并进行白细胞分类，既保证了白细胞计数值和血红蛋白测试结果的准确性，同时也提高了狗、猫等动物血液样本白细胞分类结果准确性。这种方法采用一次分血，并分别两次加溶血剂进行白细胞计数，最终以第一次计数及第二次计数 结果的比值作为判断依据，输出白细胞计数及分类结果，虽能保证大部分样本分类及计数结果的准确性，但对于血液细胞仪的重复性还有待进一步改进。
因此，现有技术还有待于改进和发展。
【发明内容】
本发明的目的在于提供一种提高白细胞分类结果准确性和计数结果重复性的方法与设备，旨在解决现有的三分类动物血液细胞分析仪检测结果不准确，重复性差的问题。
本发明的技术方案如下：
一种提高白细胞分类结果准确性和计数结果重复性的设备，其包括：用于白细胞计数的白细胞计数装置；
用于向白细胞计数装置分配血样试剂的样本血推送装置；
用于向白细胞计数装置分配溶血试剂的溶血剂推送装置；
用于分别控制样本血推送装置和溶血剂推送装置分批次向白细胞计数装置推送样本血和溶血剂的自动控制系统；
用于对白细胞计数装置进行清洗的清洗排液装置；
用于对比每次分类计数结果的准确性对比模块；
其中，所述自动控制系统先后两次控制样本血推送装置和溶血剂推送装置向白细胞计数装置推送试剂，且第一次推送的血液样本量和第二次的血液样本量相同，第二次溶血剂量小于第一次溶血剂量。
一种提高白细胞分类结果准确性和计数结果重复性的方法，其包括以下步骤：
步骤S1：向白细胞计数装置第一次分血及加入第一次溶血剂，第一次溶血剂量记作L1；
步骤S2：白细胞技术装置进行第一次白细胞计数，记作WBC1；
步骤S3：排空及清洗白细胞计数装置；
步骤S4：向白细胞计数装置第二次分血及加入第二次溶血剂，第二次溶血剂量记作L2，控制L1大于L2；
步骤S5：白细胞技术装置进行第二次白细胞计数，记作WBC2；
步骤S6：以第一次计数结果WBC1与第二次结果WBC2的比值作为判断依据，通过面积比的方式确认红细胞碎片区面积，总面积去除红细胞碎片区面积 即为第二次白细胞计数分类结果。
步骤S7：第一次计数以及第二次计数粒子数相加统计输出白细胞计数结果。
所述的方法，其以第一次计数结果WBC1与第二次结果WBC2的比值作为判断依据，通过面积比的方式确认红细胞碎片区面积的具体方法为：
WBC2/WBC1＝(Sr+Sw)/Sw
其中，Sr为第二次计数红细胞碎片区的面积，Sr/Sw＝WBC2/WBC1-1；Sw为第二次计数白细胞的面积；(Sr+Sw)为第二次计数白细胞直方图的面积。
所述的方法，其所述第二次溶血剂量L2的取值用以下步骤确定：
步骤a1：取第一溶血剂量L1值为参考，用50支样本测试得到WBC1i；
步骤a2：取上述L1值的40％-70％中的多个取值用相同标准的50支样本进行评估，先观察白细胞的分类结果，若不能实现白细胞三分类的取值则排除；能实现明显分类效果的取值作为L2的备选值，对多组L2备选值分别采用相同标准的50支样本进行计数，获取各备选值的WBC2i，将WBC2i与WBC1i比值分布在1至1.6之间最多的备选值作为第二血剂量L2的取值；
其中，i取整1至50。
本发明的有益效果：本发明通过两次向WBC池(白细胞计数池)分血及加溶血剂方法确定白细胞计数结果及分类结果，第二次计数保证了WBC分类结果的准确性。然后，将第一次白细胞计数的结果和第二次白细胞计数的结果粒子数相加，两段粒子数统计得到白细胞的计数结果,增加样本统计粒子数,提高了血液细胞分析仪WBC计数结果的重复性。该方案既保证了血液细胞分析仪白细胞分类结果准确性，又提高白细胞计数结果的重复性。
【附图说明】
图1是本发明提供的设备模块框图。
图2是本发明提供的方法流程示意图。
图3是本发明中第一次白细胞计数结果直方图示意图。
图4是本发明中第二次白细胞计数结果直方图示意图。
图5是本发明中第二次白细胞计数结果直方图中红细胞碎片区的面积和白细胞的面积的示意图。
图6是本发明中处理后的白细胞计数直方图示意图。
【具体实施方式】
为使本发明的目的、技术方案及优点更加清楚、明确，以下参照附图并举实施例对本发明进一步详细说明。
本发明的目的是提供一种提高血液细胞分析仪白细胞分类结果准确性和计数结果重复性的方法，该方案既保证了血液细胞分析仪白细胞分类结果准确性，又提高白细胞计数结果的重复性。
参见图1，本发明提供的提高血液细胞分析仪白细胞分类结果准确性和计数结果重复性的设备，主要包括模块有：
用于白细胞计数的白细胞计数装置；
用于向白细胞计数装置分配血样试剂的样本血推送装置；
用于向白细胞计数装置分配溶血试剂的溶血剂推送装置；
用于分别控制样本血推送装置和溶血剂推送装置分批次向白细胞计数装置推送样本血和溶血剂的自动控制系统；
用于对白细胞计数装置进行清洗的清洗排液装置；
用于对比每次分类结果的准确性的准确性对比模块。
其中：在本发明实施例中，所述自动控制系统先后两次控制样本血推送装置和溶血剂推送装置向白细胞计数装置推送试剂。
且，第一次推送的血液样本量和第二次的血液样本量相同，第一次的溶血剂量要足量，第二次溶血剂量小于第一次溶血剂量。
本发明提供的方法流程如图2所示，包括以下步骤：
步骤S1：向白细胞计数装置第一次分血及加入第一次溶血剂，第一次溶血剂量记作L1；
步骤S2：白细胞技术装置进行第一次白细胞计数，记作WBC1；
步骤S3：排空及清洗白细胞计数装置；
步骤S4：向白细胞计数装置第二次分血及加入第二次溶血剂，第二次溶血剂量记作L2，控制L1大于L2；
步骤S5：白细胞技术装置进行第二次白细胞计数，记作WBC2；
步骤S6：以第一次计数结果与第二次结果的比值作为判断依据，通过面积比的方式确认红细胞碎片区面积，即第一界标值，总面积去除红细胞碎片区面积即为第二次白细胞计数分类结果；
步骤S7：第一次计数以及第二次计数粒子数统计输出白细胞计数结果。
通常第一次WBC计数是完全溶解红细胞的计数结果，可直接作为WBC计数结果准确输出，根据白细胞粒子数计数公式：
白细胞计数结果白细胞有效粒子总数/(稀释比*采集时间)
其中两次白细胞计数需保证稀释比一致，采集时间可保持一致。由上述公式可知白细胞计数结果与白细胞有效粒子总数正相关。
本发明将第一次计数WBC1和第二次计数WBC2(虑除红细胞碎片后)的有效粒子总数相加，以此输出白细胞的计数结果，旨在在保证结果准确性和系统偏差不变的情况下，统计量增大，相当于减少了系统偏差对样本测试结果的影响，可以提高白细胞计数结果的重复性。
在上述实施流程中，第一次加入的溶血剂量L1足以保证使得98％以上第一次加入的动物血液样本白细胞计数结果无红细胞碎片干扰，得到第一次白细胞计数结果为WBC1，在这种状态下的白细胞直方图，如图3所示，溶血剂量L1的值可根据稀释比进行适当调整。其中，每次检测时的样本血样输入量是预先设定好的，每次相同。第一次的溶血剂量L1的选择也是预先经过多次实验调试获取的最优取值，在检测应用阶段都是已经设定好的溶血剂量，当然，本发明不排除提供具有第一次溶血剂量L1可选择的的血液分析仪。该可选择的范围也是保证使得98％以上第一次加入的动物血液样本白细胞计数结果无红细胞碎片干扰为前提的。
为了保证白细胞分类结果的准确性，第二次向白细胞计数装置中加入的溶血剂量为L2，本发明实施例中第二次溶血剂量L2小于第一次溶血剂量L1，即对白细胞计数装置中的动物血液样本采用弱溶处理的方式，使其仍存在部分的红细胞碎片区，得到第二次白细胞计数结果为WBC2，并得到第二次白细胞计数直方图，如图4所示。
L2的取值是研发实验阶段确定下来的，每次检测时我们直接取研发阶段已经确定下来的L2的最优值(定值)输入分析仪系统中进行检测即可。并不是每次检测都要进行L2的取值实验。其具体的取值方法如下：
其中，所述第二次溶血剂量L2的取值用以下步骤确定：
步骤a1：取第一溶血剂量L1值为参考，用X支样本测试得到WBC11、WBC12、WBC13……WBC1X；
步骤a2：取上述L1值的40％-70％中的多个取值用相同标准的X支样本进行评估，先观察白细胞的分类结果，若不能实现白细胞三分类的取值则排除； 能实现明显分类效果的取值作为L2的备选值，对多组L2备选值分别采用相同标准的X支样本进行计数，获取各备选值的WBC2i，将WBC2i与WBC1i比值分布在1至1.6之间最多的备选值作为第二血剂量L2的取值,i取整1至X。其中，X取值不少于50。
例如：
1)：取第一溶血剂量L1值为参考，用50支样本测试得到WBC11、WBC12、WBC13……WBC150；
2)：取上述L1值的40％、45％、50％、55％、60％、65％和70％用相同标准的50支样本进行评估，先观察白细胞的分类结果，若不能实现白细胞三分类，则不作为溶血剂量L2的取值考虑，若实现明显分类效果，例如溶血剂量L1的45％、50％、55％都能实现白细胞三分类的效果，则L2值取50支样本的WBC2i与WBC1i比值分布在1至1.6之间最多的对应溶血剂量的取值,i取整1至50。
确定溶血剂量L2值后，这时还需要确认红细胞碎片区的位置，即第一届标位置。由于动物血液样本自身的特异性，得到白细胞直方图中仍存在红细胞碎片区，将WBC1及第一次计数的WBC直方图与WBC2及第二次计数的WBC直方图进行比较可得如下关系：
WBC2/WBC1＝(Sr+Sw)/Sw
如图5所示，Sr为第二次计数红细胞碎片区的面积，Sw为第二次计数白细胞的面积，(Sr+Sw)为第二次计数白细胞直方图的面积。上述关系式可推出Sr/Sw＝WBC2/WBC1-1，由此可确定红细胞碎片区的位置，即第一界标位置。
确定第一界标位置后，第二次计数结果可以输出白细胞的分类结果，处理后的WBC直方图如图6所示。与实际镜检结果相比，保证了白细胞分类结果的准确性。对第二次白细胞计数结果作修正，即滤除掉红细胞碎片的干扰后，统计第一次和第二次计数白细胞的粒子数及采集时间，将第一次白细胞计数的结果和第二次白细胞计数的结果粒子数相加，两段粒子数统计得到白细胞的计数结果,增加样本统计粒子数,从而提高了样本WBC计数结果的重复性。
本发明通过两次向WBC池分血及加溶血剂方法确定白细胞计数结果及分类结果，第二次计数保证了WBC分类结果的准确性。然后，将第一次白细胞计数的结果和第二次白细胞计数的结果粒子数相加，两段粒子数统计得到白细胞的计数结果,增加样本统计粒子数,提高了血液细胞分析仪WBC计数结果的重复性。
应当理解的是，本发明的应用不限于上述的举例，对本领域普通技术人员来说，可以根据上述说明加以改进或变换，所有这些改进和变换都应属于本发明所附权利要求的保护范围。

Claims (11)

  1. 一种提高白细胞分类结果准确性和计数结果重复性的设备，其特征在于，包括：用于白细胞计数的白细胞计数装置；
    用于向白细胞计数装置分配血样试剂的样本血推送装置；
    用于向白细胞计数装置分配溶血试剂的溶血剂推送装置；
    用于分别控制样本血推送装置和溶血剂推送装置分批次向白细胞计数装置推送样本血和溶血剂的自动控制系统；
    用于对白细胞计数装置进行清洗的清洗排液装置；
    用于对比每次分类计数结果的准确性的准确性对比模块；
    其中，所述自动控制系统先后两次控制样本血推送装置和溶血剂推送装置向白细胞计数装置分别推送样本血和溶血剂，且第一次推送的样本血的样本量和第二次的样本血的样本量相同，第二次推送的溶血剂的剂量小于第一次推送的溶血剂的剂量。
  2. 根据权利要求1所述的设备，其特征在于，所述样本血为动物样本血。
  3. 根据权利要求2所述的设备，其特征在于，所述第一次推送的溶血剂的剂量足以保证使得98％以上第一次加入的动物样本血白细胞计数结果无红细胞碎片干扰。
  4. 一种提高白细胞分类结果准确性和计数结果重复性的方法，其特征在于，所述方法适用于权利要求1所述的设备，所述方法包括以下步骤：
    步骤S1：向白细胞计数装置第一次分样本血及第一次加入溶血剂，第一次加入的溶血剂的剂量记作L1；
    步骤S2：白细胞计数装置进行第一次白细胞计数，第一次白细胞计数结果记作WBC1；
    步骤S3：排空及清洗白细胞计数装置；
    步骤S4：向白细胞计数装置第二次分样本血及第二次加入溶血剂，第二次加入的溶血剂的剂量记作L2，控制L1大于L2，第一次的样本血的样本量和第二次的样本血的样本量相同；
    步骤S5：白细胞计数装置进行第二次白细胞计数，第二次白细胞计数结果记作WBC2；
    步骤S6：以第一次白细胞计数结果WBC1与第二次白细胞计数结果WBC2 的比值作为判断依据，通过面积比的方式确认红细胞碎片区面积，总面积去除红细胞碎片区面积即为第二次白细胞计数分类结果；
    步骤S7：第一次白细胞计数以及第二次白细胞计数粒子数相加统计输出白细胞计数结果；
    其中，以第一次白细胞计数结果WBC1与第二次白细胞计数结果WBC2的比值作为判断依据，通过面积比的方式确认红细胞碎片区面积的具体方法为：
    WBC2/WBC1＝(Sr+Sw)/Sw
    其中，Sr为第二次计数红细胞碎片区的面积；Sw为第二次计数白细胞的面积；(Sr+Sw)为第二次计数白细胞直方图的面积；
    其中，所述第二次加入的溶血剂的剂量L2的取值用以下步骤确定：
    步骤a1：取第一次加入的溶血剂的剂量L1值为参考，用50支样本测试得到WBC1i；
    步骤a2：取上述L1值的40％-70％中的多个取值用相同标准的50支样本进行评估，先观察白细胞的分类结果，若不能实现白细胞三分类的取值则排除；能实现明显分类效果的取值作为L2的备选值，对多组L2备选值分别采用相同标准的50支样本进行计数，获取各备选值的WBC2i，将WBC2i与WBC1i比值分布在1至1.6之间最多的备选值作为第二次加入的溶血剂的剂量L2的取值；
    其中，i取整1至50。
  5. 根据权利要求4所述的方法，其特征在于，所述样本血为动物样本血。
  6. 根据权利要求5所述的方法，其特征在于，所述第一次加入的溶血剂的剂量L1足以保证使得98％以上第一次加入的动物样本血白细胞计数结果无红细胞碎片干扰，得到第一次白细胞计数结果为WBC1。
  7. 一种提高白细胞分类结果准确性和计数结果重复性的方法，其特征在于，所述方法适用于权利要求1所述的设备，所述方法包括以下步骤：
    步骤S1：向白细胞计数装置第一次分样本血及第一次加入溶血剂，第一次加入的溶血剂的剂量记作L1；
    步骤S2：白细胞计数装置进行第一次白细胞计数，第一次白细胞计数结果记作WBC1；
    步骤S3：排空及清洗白细胞计数装置；
    步骤S4：向白细胞计数装置第二次分样本血及第二次加入溶血剂，第二次加入的溶血剂的剂量记作L2，控制L1大于L2，第一次的样本血的样本量和第 二次的样本血的样本量相同；
    步骤S5：白细胞计数装置进行第二次白细胞计数，第二次白细胞计数结果记作WBC2；
    步骤S6：以第一次白细胞计数结果WBC1与第二次白细胞计数结果WBC2的比值作为判断依据，通过面积比的方式确认红细胞碎片区面积，总面积去除红细胞碎片区面积即为第二次白细胞计数分类结果；
    步骤S7：第一次白细胞计数以及第二次白细胞计数粒子数相加统计输出白细胞计数结果。
  8. 根据权利要求7所述的方法，其特征在于，以第一次白细胞计数结果WBC1与第二次白细胞计数结果WBC2的比值作为判断依据，通过面积比的方式确认红细胞碎片区面积的具体方法为：
    WBC2/WBC1＝(Sr+Sw)/Sw
    其中，Sr为第二次计数红细胞碎片区的面积；Sw为第二次计数白细胞的面积；(Sr+Sw)为第二次计数白细胞直方图的面积。
  9. 根据权利要求7所述的方法，其特征在于，所述第二次加入的溶血剂的剂量L2的取值用以下步骤确定：
    步骤a1：取第一次加入的溶血剂的剂量L1值为参考，用50支样本测试得到WBC1i；
    步骤a2：取上述L1值的40％-70％中的多个取值用相同标准的50支样本进行评估，先观察白细胞的分类结果，若不能实现白细胞三分类的取值则排除；能实现明显分类效果的取值作为L2的备选值，对多组L2备选值分别采用相同标准的50支样本进行计数，获取各备选值的WBC2i，将WBC2i与WBC1i比值分布在1至1.6之间最多的备选值作为第二次加入的溶血剂的剂量L2的取值；
    其中，i取整1至50。
  10. 根据权利要求7所述的方法，其特征在于，所述样本血为动物样本血。
  11. 根据权利要求10所述的方法，其特征在于，所述第一次加入的溶血剂的剂量L1足以保证使得98％以上第一次加入的动物样本血白细胞计数结果无红细胞碎片干扰，得到第一次白细胞计数结果为WBC1。
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