| Background:We believe that erythrocyte agglutination can change the distance between red blood cells,by flow cytometry which can be accurately qualitatively and quantitatively determined.Based on this principle,we attempted to use a blood cell analyzer to qualitatively and quantitatively analyze the degree of erythrocyte agglutination.Flow cytometry(FCM)can automatically analyze and classify cells.When agglutination of red blood cells happened,cell volume is increasing,by flow cytometry which the number of cells events decreased.The equipment is characterized by standard operation,simple operation and stable results,when erythrocyte agglutination occurs,a number of small red blood cells of group will become a large red blood cells,by the drop of the count value of red blood cells,in the automatic blood cell analyzer will be directly reflected.In this study,the RBC agglutination can be qualitatively and quantitatively detected by flow cytometry can be proved.,by automatic blood cell analyzer,the degree of erythrocyte agglutination can be further quantitatively determined.By anti-erythrocyte antibodies,the difference in the degree of erythrocyte agglutination between healthy and immune diseases can be demonstrated.In this study,by collecting samples from ANA positive patients,a methodology for quantitative detection of erythrocyte agglutination by automatic blood cell analyzer was established,which can quantitatively detect the agglutination of anti-erythrocyte antibodies in ANA positive samples,a new research idea for clinical practice will be opened.Methods:1.Between red blood cells of two blood groups A and B and serum of two blood groups B and A respectively,the agglutination experiment was conducted.The conditions were as follows: the concentration of red blood cells was fixed,the serum was diluted by multiple ratios.By warm bath at 37 ℃ for 30 minutes,the constant temperature was established.2.Respectively with two red blood cells of blood type A,B and two kinds of blood serum B,A warm bath at 37 ℃ for 30 minutes,the agglutination of the results observed by the naked eye on the glass sheet was recorded and then individually determined by automatic blood cell analyzer,ensure each tube samples before sample already blending.3.Agglutination index AGI was established,AGI=RBC count in the disease group/RBC count in the control group.(The result preserves two decimal places).The AGI value by comparing it with the agglutination reaction in the sera was obtained.The AGI value was used to further explain the agglutination of anti-erythrocyte antibodies in the blood samples of ANA-positive patients,and the clinical application value of anti-erythrocyte antibodies in autoimmune diseases was further proved.4.Clinical specimens positive for ANA were collected,and serum was isolated,stored at-20℃ for later use.Preparation of type O anticoagulant whole blood as instructs the cell,and ANA positive serum specimens under different concentration,temperature of 37 ℃ bath for 30 minutes,one by one after blending determination by automatic blood cell analyzer,by illness and health specimens were similar in age,the same blood type,contrast experiment under the condition of the same reaction conditions,a new testing agglutinate phenomenon of methodology was established.Results:1.By flow cytometry,which can be used for the determination of erythrocyte agglutination by observing the appearance of erythrocyte agglutination.It is proved that the flow cytometry can indicate the agglutination of erythrocytes under the condition of fixed volume parameters,and a new method for detecting the agglutination of erythrocytes by flow cytometry has been established initially.2.When the volume parameters of the flow cytometry were fixed,the value of ABS3 showed a linear relationship with the dilution ratio of red blood cells,indicating that with the increase of the dilution ratio of red blood cells,the value of ABS 3 increased and the degree of agglutination decreased.The quantitative detection of red blood cell agglutination could be achieved,which the traditional glass slide method was highly correlated.3.The samples of healthy people and the serum of ANA-positive patients were used to agglutinate with erythrocytes.The quantitative erythrocyte value can be decreased regularly on the automatic blood cell analyzer.Low levels of anti-O erythrocyte antibodies are demonstrated in patients with ANA-positive autoimmune disease or suspected autoimmune disease.Conclusion:1.Flow cytometry can be used for qualitative and quantitative detection of erythrocyte agglutination.2.The method of quantitative determination of erythrocyte agglutination by automatic blood cell analyzer have been established.3.Anti-erythrocyte antibodies were found in autoimmune diseases by method of automatic blood cell analyzer. |
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