Peruzzi Benedetta, Guerrieri Serena, Biagioli Tiziana, Lanzilao Luisa, Pratesi Sara, Bencini Sara, Statello Marinella, Carraresi Alessia, Stefanelli Stefania, Tonelli Martina, Brogi Marco, Capone Manuela, Mazzoni Alessio, Gelli Anna Maria Grazia, Fanelli Alessandra, Caporale Roberto, Annunziato Francesco
Flow Cytometry Diagnostic Centre and Immunotherapy (CDCI), AOU Careggi, Largo Brambilla, 3, 50134, Florence, Italy.
Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
Pract Lab Med. 2024 May 16;40:e00401. doi: 10.1016/j.plabm.2024.e00401. eCollection 2024 May.
Recently, a flow cytometric (FC) based test has been developed for detection of circulating fetal cells to replace the less accurate and reproducible Kleihauer-Betke test.FC test is easier to perform, it can distinguish the origin of fetal cells, but it is expensive and available in highly specialized laboratories. We evaluated the introduction of high-performance liquid chromatography (HPLC) approach as initial screening to identify patients who need an additional FC test to better discriminate the nature of haemoglobin-F (HbF) positive cells.
Blood samples from 130 pregnant women suspected to have fetomaternal haemorrhage were analysed with HPLC and FC methods. The cut-off for HbF HPLC concentration was calculated. Statistical analyses for the evaluation of HPLC as a screening method were performed. The positivity cut-off of HbF to be used as decision-making value to continue the investigation was calculated.
An excellent agreement (R > 0.90) was observed between the percentage of HbF obtained by HPLC and the percentage of fetal cells detected by FC. Results obtained from each assay were compared to define the HPLC threshold below which it is not necessary to continue the investigations, confirming the maternal nature of the HbF positive cells detected. Our study demonstrated that a cut-off of 1.0 % HbF obtained by HPLC was associated with the lowest rate of false negative results in our patient cohort.
This study provides a new FMH investigation approach that possibly leads to a reduction in times and costs of the analysis.
最近，已开发出一种基于流式细胞术（FC）的检测方法来检测循环中的胎儿细胞，以取代准确性和可重复性较低的克莱豪尔-贝特克试验。FC检测操作更简便，能够区分胎儿细胞的来源，但成本高昂，且只有在高度专业化的实验室才能进行。我们评估了采用高效液相色谱（HPLC）方法作为初始筛查手段，以识别那些需要进一步进行FC检测以更好地鉴别血红蛋白F（HbF）阳性细胞性质的患者。
对130名疑似发生胎儿-母体出血的孕妇的血样进行HPLC和FC分析。计算HbF HPLC浓度的临界值。对HPLC作为筛查方法进行评估的统计分析。计算用作继续调查决策值的HbF阳性临界值。
HPLC测得的HbF百分比与FC检测到的胎儿细胞百分比之间观察到极好的一致性（R > 0.90）。比较每种检测方法得到的结果，以确定HPLC阈值，低于该阈值则无需继续调查，从而确认检测到的HbF阳性细胞的母体性质。我们的研究表明，HPLC测得的HbF临界值为1.0%时，在我们的患者队列中假阴性结果率最低。
本研究提供了一种新的胎儿-母体出血调查方法，可能会减少分析的时间和成本。
