| Part One : The evaluation of the accuracy of platelet count by XN9000 hematology analyzer.1)Effects of sample preservation temperature and preservation time on the detection of PLT and IPF.Objective: To observe the effect of different preservation temperatures and preservation times on the detection results of reticulated platelet.Methods: Twenty healthy subjects were randomly selected and enrolled in this study,and 2ml EDTA-K2 coagulation vein blood was extracted from every subject.The samples were used to detect IPF by XN9000 automated hematology analyzer immediately,or after the preservation in 4? refrigerator for 24,48 or 72 hours,respectively(four time points).In addition,20 inpatients were also randomly selected,and 2ml EDTA-K2 coagulation vein blood was also extracted from these patients.The samples were used to detect IPF every two hours under room temperature,and the corresponding results were observed and recorded.Results:After 24-hour preservation in 4? refrigerator,the means of IPF and IPA showed a significant difference with that detected within 24 hours(P=0.003,0.006).The samples preserved under room temperature showed detection results stable within 4hours.Conclusion: The detection of reticulated platelets in EDTA-K2 coagulation whole blood was stable within 4 hours under room temperature;and the blood samples were not suitable for reticulated platelets detection after one-day preservation in 4? refrigerator.2)Accuracy evaluation of low platelets count using XN9000(Compared with the results of XE2100 hematology analyzer).Objective: To investigate the property of low-valued platelets counts by the new fluorescence method(PLT-F)on Sysmex XN9000 automatic hematology analyzer.Methods: 50 routine blood samples,which PLT counts were below 50×109/L detected by PLT-I method on Sysmex XE2100,were selected in this study.Platelet counts of these samples were also analyzed with the parameter profile “CBC+RET+PLT-F” on Sysmex XN9000 automatic hematology analyzer.The properties of low platelets counts by Sysmex XN9000 automatic hematology analyzer were evaluated from some aspects,including precision and different instruments.And the results were also compared with flow cytometry analysis of CD61-Immuno PLT according to ICSH recommendatory methods.Results: Compared with the impedance method(PLT-I),the optical method(PLT-O),the repeatability of PLT-F method(New fluorescence method on Sysmex XN9000 automatic hematology analyzer)was best,with the highest precision(CV<4%);Though all of the three methods showed good correlation with the results of flow cytometry,the results of PLT-F show the strongest correlation with CD61-immuno PLT of flow cytometry(r=0.9636,P<0.0001).Conclusion: The new fluorescence method(PLT-F)of Sysmex XN9000 automatic hematology analyzer showed high precision and good repeatability for low platelets counts,and it showed significantly positive correlation with a reference method,which showed that the results were accurate and reliable.When the PLT counts was lower detected by XE2100,it was recommended to be rechecked by PLT-F method of XN9000 automatic hematology analyzer.And it could also be detected on flow cytometry if necessary.Part Two: To establish the reference interval of reticulated platelet percentage(IPF)and absolute value(IPA).1)To establish the reference interval of reticulated platelet percentage and its absolute value in healthy adults.Objective: To establish the reference interval of reticulated platelet percentage(IPF)and absolute value(IPA)in healthy adults of our area.Methods: The blood samples were analyzed by XN9000 automatic hematology analyzer in 2179 physical examination subjects.There were 1138 males and 1041 females,with the median age of 33 years(range 18-92 years).Inclusive criteria: 1)The results of general physical examination did not show obvious abnormality.And the results of ECG,chest X-ray fluoroscopy,ultrasound examination were also normal.2)The results of detection indicators showed that the subjects had no obvious symptoms of infection and anemia performance.White blood cell(WBC)<10×109/L;Hemoglobin(HB)male>120g/L;female >110g/L;PLT>125×109/L.3)The results of biochemical indexes were normal.Fasting peripheral blood were collected in the early morning from all the subjects.The samples were 2ml EDTA-K2 coagulation blood,which was completed the detection within 2 hours.And the data were analyzed by SPSS 19.0 software.Results: In adults,the IPF reference interval:(0.7-8.4):%;IPA reference interval:(2.9 ~ 16.9)×109/L.There was the significant difference of PLT count between adult males and females(P=0.032).There wasno significant difference of IPF and IPA between adult males and females(P=0.232,0.511,respectively).And the values of IPA and IPF showed significant differences between different age groups(P = 0.021,0.010,respectively).Conclusion:This study established the reference interval of reticulated platelet percentage and its absolute value by instrument methods in healthy adults of our area,in order to elevate the clinical application value of reticulated platelets detection.2)To establish the reference interval of reticulated platelet percentage and absolute value in umbilical cord blood of full term neonates.Objective: To establish the reference interval of reticulated platelet percentage(IPF)and absolute value(IPA)of umbilical cord blood.Methods: The umbilical cord blood were collected from 153 healthy full-term newborn infants,and the reticulated platelet percentage and absolute value were detected.The samples were 2ml EDTA-K2 coagulation blood,which was completed the detection within 2 hours.And the data were analyzed by SPSS 19.0 software.Results: In umbilicalcordblood,the reference interval of IPF was(0.9-4.8)%,and the reference interval of IPA was(3.1-13.5)×109/L.There were statistically significant differences of these above mentioned parameters between umbilical cord blood and peripheral blood of adults(all P value was <0.001,LT,IPF,IPA,results of Mann-Whitney U test).Conclusion:This study firstly established the reference interval of reticulated platelet percentage and its absolute value by instrument method in umbilical cord blood of full term neonates in China,in order to further provide the basis for the clinical application value of reticulated platelets related parameters in neonatal and children thrombocytopenia diseases.Part 3: Study on the clinical application of reticulated platelets.1)The application of reticulated platelets in thrombocytopenic diseases.Objective: To investigate the correlation of reticulated platelet and platelet parameters,and their clinical application value in the patients with thrombocytopenia.Methods: ITP patients(32 cases),AA patients(40 cases),acute leukemia patients combined with thrombocytopenia(80 cases),and MDS patients(20cases)were collected from outpatients and inpatients.2ml EDTA-K2 coagulation blood was collected form every patient,and reticulated platelets(IPF)and platelet related parameters(PLT,PDW,PCT,MPV,P-LCR)were detected by low-valued platelets dedicated channel(PLT-F)of Sysmex XN9000 automatic hematology analyzer,respectively.The results of patient groups were compared with healthy control subjects(544 cases).The correlations between IPF and other platelet related parameters were analyzed using data from healthy control group.Results: In healthy control group,the results of correlation analysis indicated that IPF was negatively associated with PLT count and thrombocytocrit(PCT)(r=-0.427,-0.178,respectively).IPF showed positively associated with PDW,MPV,and P-LCR(r=0.802,0.782,0.365,respectively).The value of IPF in ITP group(13.91±9.22%)was obviously higher than healthy control group(4.92±2.46%),and the difference was significant(P<0.001);the value of IPF in AL group(3.91±3.03%)was obviously lower than healthy control group,and the difference was significant(P<0.05).The value of IPF in AA group and AL group was obviously lower than healthy control group,and the difference was significant(P<0.001).In each disease group,the value of PLT was obviously lower than that of healthy control group(all P value<0.001,for ITP group,AA group and AL group,respectively;P<0.05 for MDS group).The mean platelet volume(MPV)of ITP group was higher than that of healthy control group,while MPV value in the AA group was lower than that of healthy control group.And there was no significant difference in the platelet histogram distribution width(PDW)between every two groups(all p value > 0.05).In ITP group,the IPF values were compared at time points before and after the treatment.After treatment,PF% and MPV value significantly decreased,and PLT significantly elevated in patients of ITP group(P < 0.05).There was no significant difference of IPA value between time points before and after treatment(P>0.05).Conclusion: Compared with healthy control subjects,there were significant differences of reticulated platelets and platelets related parameters in patients with thrombocytopenia.And the above mentioned parameters also showed significant difference in thrombocytopenia patients with various etiologies.Through the comprehensive analysis of reticulated platelets and platelet related parameters,we could make a better evaluation of the etiology identification and therapeutic effect in patients with thrombocytopenia.2)Monitoring of PLT recovery after hematopoietic stem cell transplantation(HSCT): Objective: To search for the suitable cut off value of IPF for predicting platelet recovery 2 days after HSCT.Methods: 29 patients who completed the treatment of hematopoietic stem cell transplantation in our hospital were traced,and we routinely detect various platelet parameters in peripheral blood after HSCT treatment by Sysmex XN9000 automatic hematology analyzer.ROC curve was used to analyze the cut off value of IPF in 29 patients received HSCT treatment,in order to predict PLT recovery 2 days after HSCT.Results: 1.In all cases,the platelet counts reduce to the minimum at the mean of 9.1 days(range: 6-12 days)after HSCT.And the platelets recovery occurred at the mean of 14.7 days(range 10-31 days).The recovery about the absolute value of reticulated platelets occurred at the mean of 11.5days after HSCT(range 8-24 days).The recovery of reticulate platelets ratio(IPF)occurred at the mean of 10.3 days(range: 8-23 days)after HSCT.All the cases could be observed the elevation of IPF and IPA before PLT recovery.The recovery of IPF was about 5.1 days(range: 2-9 days)earlier than that of PLT,and the recovery of PLT was about 3.9 days(range: 1-7 days)earlier than that of IPA.2.In patients received bone marrow transplant(17 cases): The platelet reconstruction occurred at the median of 15.5 days(range 10-28 days)after transplantation.And the reconstruction of absolute value of reticulated platelets(IPA)occurred at the median of 12.2 days(range 9-15 days)after transplantation.In addition,the reconstruction of reticulated platelets ratio(IPA)occurred at the median of 11.5 days(range: 10-13 days)after transplantation.The time of reticulated platelets reconstruction was about 4 days earlier than the time of PLT reconstruction,and the time of reticulated platelets absolute value construction was about three days earlier than PLT reconstruction.3.In patients received peripheral blood stem cell transplantation treatment(11 cases)(autotransplantation: 5 cases,allograft: 6 cases).The platelet reconstruction occurred at the mean of 12 days(range:10-18 days),and the reconstruction of absolute value of reticulated platelets(IPA)occurred at the mean of 10.6 days(range 8-14 days)after transplantation.In addition,the reconstruction of reticulated platelets ratio(IPA)occurred at the mean of 9.1 days(range: 8-12 days)after transplantation.If subdivided,it could be observed that the time of platelets reconstruction in patients received autotransplantation(median 11.2 days,range: 10-13 days)was earlier than that of patients received allograft transplantation(median: 13.1 days,range: 10-18days).The results were listed in Table 16.Overall,compared with bone marrow stem cell transplantation,the recovery time of platelet and reticulated platelet was about 3days earlier than that of peripheral blood stem cell transplantation,especially in patients received the treatment of autologous hematopoietic stem cell transplantation.4.The result of ROC curve analysis showed that when 3.5% was used to be the cut off value of IPF,then the sensitivity was 78%,the specificity was 89%,the positive predictive value was 91% for its predictive value of PLT reconstruction.Conclusion: 1.In patients treated by stem cells transplantation,the reconstruction of reticulated platelet percentage was 4 days earlier than PLT reconstruction.And the reconstruction of reticulated platelet absolute value was about3 days earlier than PLT reconstruction.2.Though ROC curve analysis,it was confirmed that the cut off value of IPF was3.5%,which could be used to predict PLT reconstruction after hematopoietic stem cell transplantation within 2 days after IPF elevation.Through establishing this cut off value,it was expected to help reduce treatment of prophylactic platelet transfusion,further reduce the transfusion related risk and patient's economic burden. |
PDF Full Text Request