未成熟血小板检测方法的比较

Comparison of testing methods for reticulated platelets

目的分析比较检测未成熟血小板的方法学差异，并调查本地区正常参考区间，为实验室开展未成熟血小板检测提供依据。方法分析使用聚甲基曝嗪（PO）染色的全自动血细胞分析仪和用噻唑橙（TO）染色的传统流式细胞分析仪检测未成熟血小板的批内精密度、携带污染率等指标，分析同一仪器在不同时间检测的未成熟血小板百分比的差异，比较两种方法的性能，检测健康体检人群（n=336）的未成熟血小板百分比，分析两种方法的相关性，统计全自动血细胞分析仪的参考区间。结果全自动血细胞分析仪的变异系数（CV）为16．2％、携带污染率为4．6％；流式细胞分析仪的最少CV为35．1％、携带污染率为4．8％。样本放置30rain、2h、4h后，全自动血细胞分析仪检测未成熟血小板，差异无统计学意义（P〉0．05）；TO染色后30min、1h、 2h用传统流式细胞分析仪检测的未成熟血小板，差异有统计学意义（P〈0．01）。两种方法检测结果有相关性（n=0．923，P〈0．01）。全自动血细胞分析仪检测值的正常参考区间为1．0％～7．5％；流式细胞分析仪检测值的正常参考区间为3．O％-10．5％。结论全自动血细胞分析仪法比传统流式细胞分析仪法操作简单，结果稳定性好，是检测未成熟血小板的优选方法。

Objective To compare the accuracy, stability and sample cross-contamination of two independent methods of detecting the percentage of reticulated platelets in peripheral blood and establish a local normal reference range so as to provide methodological rationales in clinical laboratory. Methods The percentages of reticulated platelets in peripheral blood of a healthy population were measured by Sysmex XE- 5000 blood cell analyzer with polymethyl oxazine staining and flow cytometer with thiazole orange staining respectively. The correlation between the results of two methods was analyzed by Spearman＇s nonparametric correlation. Information about stability was obtained from measurements of the percentages of reticulated platelets in peripheral blood at designated timepoints. The analyses of accuracy, sample cross contamination and local normal reference range were performed routinely. Results The coefficient of variation （CV） of data was lower （ 16. 2% ） than that from flow cytometer （35.1%）. The sample cross-contaminations of two methods were the same at around 5%. The percentage of reticulated platelets in peripheral blood was stable and consistent whereas the results of flow cytometer fluctuated at different timepoints within 4 h after blood sampling. The correlation of results obtained from two methods was significant （P 〈 O. O1, r2 = 0. 923 ）. The local normal reference range was 1.0% - 7.5% for Sysmex XE-5000 versus 3.0% - 10. 5% for flow cytometer. Conclusions Fully automatic blood cell analyze is more advanced than flow cytometer for its simple operation and stable data. And the former is an ideal first-choice for detecting the percentage of reticulated platelets in peripheral blood.