Department of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Warsaw, Poland.
Department of Pediatric Hematology and Oncology, Medical University of Warsaw, Warsaw, Poland.
Cytometry B Clin Cytom. 2018 Jan;94(1):189-195. doi: 10.1002/cyto.b.21511. Epub 2017 Feb 1.
Osmotic fragility test (OFT) is widely considered as a sensitive indicator of red blood cells' sensitivity to the hypotonic solution. It is often used as a screening test for the diagnosis of hereditary spherocytosis (HS). Nowadays, the osmotic fragility test based on flow cytometric analysis (FCM OF) is widely used in laboratory practice. The purpose of this study was to optimize the assay sensitivity and to validate its clinical application in the diagnostic screening of childhood anemias.
The study was conducted on 175 children suffering from various types of anemia (including 30 children with proven hereditary spherocytosis, HS) and 16 healthy subjects. All children were aged between 3 months and 17 years, including 94 boys and 97 girls. FCM OF was performed on every subject according to two different analysis time patterns (hemolysis was analyzed for 214 or 300 s) using Cytomics FC500 flow cytometer.
Significant higher sensitivity was demonstrated by the tests carried out according to the longer analysis time pattern (90.0 vs. 83.33%). The level of specificity of both the analysis patterns was similar. When an extended analysis time was used, the percentage of red cell survival levels in HS patients were significantly lowered compared to the same cases analyzed with shorter incubation times and all other non-HS anemic cases (9.31 ± 4.69 vs. 35.59 ± 15.30%, P < 0.05). During the shorter analysis time, the values obtained were 13.76 ± 7.92% for HS and 48.18 ± 19.04% for non-HS, P < 0.0001. The 300-s test is very useful in distinguishing thalassemia patients from patients with other types of anemias (94.74% sensitivity and 90.12% specificity) and provided the values of remaining red blood cells as 70.46 ± 12.29% for thalassemia and 27.16 ± 13.01% for nonthalassemia subjects, P < 0.0001.
Flow cytometric osmotic fragility test with a longer (300-s) analysis time demonstrated an increased sensitivity in detecting HS in anemic children. © 2017 International Clinical Cytometry Society.
渗透脆性试验（OFT）被广泛认为是红细胞对低渗溶液敏感性的敏感指标。它通常用作遗传性球形红细胞增多症（HS）诊断的筛选试验。如今，基于流式细胞术分析的渗透脆性试验（FCM OF）已广泛应用于实验室实践。本研究旨在优化检测敏感性，并验证其在儿童贫血症诊断筛查中的临床应用。
本研究共纳入 175 名患有各种类型贫血的儿童（包括 30 名遗传性球形红细胞增多症患儿，HS）和 16 名健康受试者。所有儿童年龄在 3 个月至 17 岁之间，包括 94 名男孩和 97 名女孩。使用 Cytomics FC500 流式细胞仪，对每位受试者进行两种不同的分析时间模式（溶血分析 214 或 300 s）的 FCM OF。
采用较长分析时间模式进行的检测显示出更高的灵敏度（90.0%对 83.33%）。两种分析模式的特异性水平相似。当使用扩展的分析时间时，HS 患者的红细胞存活率水平明显低于使用较短孵育时间和所有其他非 HS 贫血病例的水平（9.31±4.69%对 35.59±15.30%，P<0.05）。在较短的分析时间内，HS 患者的检测值为 13.76±7.92%，非 HS 患者的检测值为 48.18±19.04%，P<0.0001。300 s 检测在区分地中海贫血患者和其他类型贫血患者方面非常有用（敏感性 94.74%，特异性 90.12%），并提供了地中海贫血患者的剩余红细胞值为 70.46±12.29%，非地中海贫血患者的剩余红细胞值为 27.16±13.01%，P<0.0001。
流式细胞术渗透脆性试验，采用较长（300 s）分析时间，可提高对贫血儿童 HS 的检测灵敏度。 © 2017 年国际临床细胞学会。
