红细胞内疟原虫干扰网织红细胞计数纠正方法的研究

Research on the calibration method of reticulocyte count interfered by plasmodium in red blood cells

目的:解决血液标本中红细胞内疟原虫对网织红细胞(Reticulocyte,Ret)计数结果的干扰问题,对网织红细胞检测结果进行校准。方法:通过两种不同的染色方法,一部分标本用新亚甲蓝或煌焦油蓝染液对血液常规标本进行活体染色后,采用手工或血液细胞分析仪对其进行网织红细胞计数;另一部分制成血涂片,使用瑞姬氏染色后,对含疟原虫的红细胞进行计数(与Ret的手工计数方法相同),含疟原虫红细胞%=1000个红细胞中含有疟原虫的红细胞个数/10。Ret的校正公式:Ret#(绝对数)校正值(×1012/L)=校正前Ret#-红细胞绝对数×含疟原虫的红细胞%,或Ret%(百分率)校正值=校正前Ret%-含疟原虫的红细胞%。结果:红细胞内疟原虫含RNA与网织红细胞内残存的RNA相似,经活体染色后,仪器和手工计数难以分辨,使Ret不同程度假性增高,10例疟疾患者Ret(%)的均值±标准差(±s)校正前后分别为9.86±13.89和6.63±10.93,Ret#(×1012/L)校正前后分别为0.35±0.47和0.09±0.11。疟疾患者的Ret结果受干扰的程度与含疟原虫红细胞数量有关,含疟原虫红细胞的百分率越高,干扰越大。结论:红细胞内疟原虫常可引起Ret假性增高,且增高程度极大,需要对Ret的结果进行校正。该方法具有简便、实用和科学的特点,通过该校正方法对红细胞内疟原虫干扰Ret检测的结果的校正,能够使仪器检测Ret结果更加准确可靠。

Objective：To solve the problem that plasmodium in red blood cells（RBC）interfere the reticulocyte（Ret）count and to calibrate the results.Methods：Samples were stained by two different dyeing methods.One method was that blood sample stained by methylene blue or brilliant cresol blue,then reticulocytes were counted by blood cell analyzer or manually.The other was that blood smears were stained by Wright-Giemsa and RBCs contained with plasmodium were counted with plasmodium in 1000RBCs/10.Calibration formula could be balibrated Ret#（×1012/L）=Ret# before calibration-RBC#×Percentage of RBCs contained with plasmodium,or calibrated Ret%=Ret% before calibration-Percentage of RBCs contained with plasmodium.Results：RNAs of plasmodium in RBC was similar with those in reticulocytes.It was difficult to distinguish RBCs with plasmodium from reticulocytes with ananlyzer or manually,which would result in a false increase in Ret count.Before and after calibration,x±s of Ret% in 10 patients with malaria were 9.86±13.89 and 6.63±10.93,respectively.While x±s of Ret# were 0.35±0.47 and 0.09±0.11,respectively.Ret count in malaria patients was relevant with the number of RBCs with plasmodium.The higher the percentage of RBCs with plasmodium was,the more it could interfere the reticulocyte count.Conclusion：Plasmodium inRBCs would cause a false increase reticulocyte count and should calibrate the Ret results before report.Calibrated Ret count results with convenient,practical method and make manual or automated Ret count more accurate and reliable.