采用高效液相色谱法和流式细胞术相结合的方法分析胎儿产妇出血评估中的 HbF

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY Practical Laboratory Medicine Pub Date : 2024-05-01 DOI:10.1016/j.plabm.2024.e00401
Benedetta Peruzzi , Serena Guerrieri , Tiziana Biagioli , Luisa Lanzilao , Sara Pratesi , Sara Bencini , Marinella Statello , Alessia Carraresi , Stefania Stefanelli , Martina Tonelli , Marco Brogi , Manuela Capone , Alessio Mazzoni , Anna Maria Grazia Gelli , Alessandra Fanelli , Roberto Caporale , Francesco Annunziato
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引用次数: 0

摘要

导言最近,一种基于流式细胞术(FC)的检测方法被开发出来,用于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雷豪尔-贝特克检测方法。FC检测方法更易于操作,可区分胎儿细胞的来源,但其价格昂贵,且只能在高度专业化的实验室中使用。我们评估了采用高效液相色谱法(HPLC)进行初步筛查的方法,以确定哪些患者需要进行额外的 FC 检验,从而更好地鉴别血红蛋白-F(HbF)阳性细胞的性质。计算出 HbF HPLC 浓度的临界值。对高效液相色谱法作为筛查方法的评估进行了统计分析。结果 HPLC 检测到的 HbF 百分比与 FC 检测到的胎儿细胞百分比之间的一致性非常好(R2 > 0.90)。通过比较每种检测方法得出的结果,确定了 HPLC 临界值,低于该临界值则无需继续检测,从而确认了检测到的 HbF 阳性细胞的母体性质。我们的研究表明,在我们的患者队列中,HPLC 检测出的 HbF 临界值为 1.0%,假阴性结果率最低。
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HPLC and flow cytometry combined approach for HbF analysis in fetomaternal haemorrhage evaluation

Introduction

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.

Methods

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.

Results

An excellent agreement (R2 > 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.

Conclusions

This study provides a new FMH investigation approach that possibly leads to a reduction in times and costs of the analysis.

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来源期刊
Practical Laboratory Medicine
Practical Laboratory Medicine Health Professions-Radiological and Ultrasound Technology
CiteScore
3.50
自引率
0.00%
发文量
40
审稿时长
7 weeks
期刊介绍: Practical Laboratory Medicine is a high-quality, peer-reviewed, international open-access journal publishing original research, new methods and critical evaluations, case reports and short papers in the fields of clinical chemistry and laboratory medicine. The objective of the journal is to provide practical information of immediate relevance to workers in clinical laboratories. The primary scope of the journal covers clinical chemistry, hematology, molecular biology and genetics relevant to laboratory medicine, microbiology, immunology, therapeutic drug monitoring and toxicology, laboratory management and informatics. We welcome papers which describe critical evaluations of biomarkers and their role in the diagnosis and treatment of clinically significant disease, validation of commercial and in-house IVD methods, method comparisons, interference reports, the development of new reagents and reference materials, reference range studies and regulatory compliance reports. Manuscripts describing the development of new methods applicable to laboratory medicine (including point-of-care testing) are particularly encouraged, even if preliminary or small scale.
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