Research

12 pages, 891 KiB

Open AccessArticle

Prospective Evaluation of Fetal Hemoglobin Expression in Maternal Erythrocytes: An Analysis of a Cohort of 345 Parturients

by Laurence Blain, Christian Watier, Xiaoduan Weng, Andre Masse, Marie-Josée Bédard, Nazila Bettache, Florence Weber, Michele Mahone, Stéphanie Forté, Vincent-Philippe Lavallée, Pierre-Olivier Gaudreau, Michael J. Newmarch and Denis Soulières

Diagnostics 2023, 13(11), 1873; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics13111873 - 27 May 2023

Viewed by 1122

Abstract

It is believed that fetal hemoglobin (HbF) expression in adults is largely genetically regulated. The increased expression of HbF in pregnancy has been reported in a small number of articles. Different mechanisms have been proposed, but the description of HbF expression during pregnancy [...] Read more.

It is believed that fetal hemoglobin (HbF) expression in adults is largely genetically regulated. The increased expression of HbF in pregnancy has been reported in a small number of articles. Different mechanisms have been proposed, but the description of HbF expression during pregnancy remains unclear. The objectives of this study were to document HbF expression during peri and postpartum periods, confirm its maternal origin, and assess clinical and biochemical parameters potentially associated with HbF modulation. In this observational prospective study, 345 pregnant women were followed. At baseline, 169 had HbF expression (≥1% of total hemoglobin) and 176 did not have HbF expression. Women were followed at the obstetric clinic during their pregnancy. Clinical and biochemical parameters were measured at each visit. Analyses were made to determine which parameters had a significant correlation to HbF expression. Results show that HbF expression of ≥1% during peri and postpartum periods in pregnant women without influencing comorbidities is at its highest peak during the first trimester. In all women, it was proven that HbF was of maternal origin. A significant positive correlation between HbF expression, βeta-human chorionic gonadotropin (β-HCG), and glycosylated hemoglobin (HbA1c) was present. A significant negative association between HbF expression and total hemoglobin was found. HbF expression induction during pregnancy is probably associated with an increase in β-HCG and HbA1C, and a decrease in total hemoglobin, which could temporarily reactivate the fetal erythropoietic system. Full article

(This article belongs to the Special Issue Diagnosis for Hematologic Disorders)

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12 pages, 459 KiB

Open AccessArticle

Predictors of Remission in Severe Childhood Immune Thrombocytopenia

by Chao-Neng Cheng, Yuan-Ning Yang, Yun-Hsuan Yeh, Li-Wen Chen, Jiann-Shiuh Chen and Yung-Chieh Lin

Diagnostics 2023, 13(3), 341; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics13030341 - 17 Jan 2023

Cited by 1 | Viewed by 2585

Abstract

Childhood immune thrombocytopenia (ITP; platelet count < 100 × 10⁹/L) is the most common bleeding disorder in children. A total of 3–5% of children with ITP face a greater risk of bleeding, resulting in significant morbidity and mortality. Childhood ITP is [...] Read more.

Childhood immune thrombocytopenia (ITP; platelet count < 100 × 10⁹/L) is the most common bleeding disorder in children. A total of 3–5% of children with ITP face a greater risk of bleeding, resulting in significant morbidity and mortality. Childhood ITP is often benign and self-limited; however, children with severe ITP (platelet count < 30 × 10⁹/L) require investigation and monitoring. In addition, 20% of ITP patients may not go into remission (platelet counts < 100 × 10⁹/L by 12 months after diagnosis) and may develop chronic ITP. The early identifying predictors associated with the resolution of severe ITP at the time of diagnosis may be helpful for family guidance. However, there is still controversy about the associations between the clinical factors at the time of initial diagnosis and the definitions of disease remission assessed at different timepoints after diagnosis. This retrospective study aimed to analyze the shared clinical factors among the disease remission definitions at three arbitrarily set timepoints—3, 6, and 12 months after diagnosis. This study retrieved records for hospitalized children aged under 18 years and diagnosed with ITP from the hospital registry in a tertiary university hospital. Clinical variables were recorded by reviewing the medical records with structured data entry for ITP admission. The serial follow-up platelet counts within 12 months after diagnosis were recorded. The times of ITP remission were identified by experienced pediatric hematologists. Patients with mild-form ITP (platelet counts ≥ 30 × 10⁹/L) at diagnosis or who were lost to follow-up within 3 months were excluded. From 1988 to 2019, 546 children were enrolled, and a total of 497 children with severe ITP were included in the further analysis. In total, one (0.2%) died of an intracranial hemorrhage, 363 (73.2%) children went into remission at 3 months, 40 (8.1%) went into remission between 6 and 12 months, and 104 (20.9%) developed chronic ITP. The shared significant predictors for remission by the third, sixth, and twelfth months included pre-adolescent age (<10 years) at diagnosis, abrupt onset (duration of symptoms prior to admission ≤ 2 weeks), and speedy recovery (platelet count > 100 × 10⁹/L at 1 month post diagnosis). ITP patients with positive viral serology tests or vaccination within 4 weeks had trends of delayed remission. In conclusion, diagnosis before preadolescent age, abrupt onset, and speedy recovery may share favorable factors for the remission of childhood ITP assessed at different timepoints. Full article

(This article belongs to the Special Issue Diagnosis for Hematologic Disorders)

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12 pages, 1520 KiB

Open AccessArticle

Evaluation of the Analytical Performances of the Biolabo SOLEA 100 Optical Coagulometer and Comparison with the Stago STA-R MAX Analyser in the Determination of PT, APTT, and Fibrinogen

by Pierangelo Bellio, Simonetta De Angelis, Alessandra Piccirilli, Giulio Di Michele, Remo Barnabei, Gianfranco Amicosante, Mariagrazia Perilli and Giuseppe Celenza

Diagnostics 2023, 13(1), 85; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics13010085 - 28 Dec 2022

Viewed by 1464

Abstract

Introduction. The Biolabo Solea 100 is a fully automated coagulation analyser using an optical system to detect coagulation designed to meet the needs of small- and medium-sized laboratories. This study aimed to evaluate the analytical performance in terms of bias, precision, and interference [...] Read more.

Introduction. The Biolabo Solea 100 is a fully automated coagulation analyser using an optical system to detect coagulation designed to meet the needs of small- and medium-sized laboratories. This study aimed to evaluate the analytical performance in terms of bias, precision, and interference of the Biolabo Solea 100 coagulometer under routine laboratory conditions. In addition, a comparison was made with Stago STA-R MAX. Materials and Methods. Imprecision and bias were evaluated for activated partial thromboplastin time (APTT), fibrinogen (FIB), and prothrombin time (PT) at the medical decision levels. The results of 200, 181, and 206 plasma samples for APTT, FIB, and PT, respectively, were compared with those obtained by Stago STA-R MAX. In addition, the interference level of bilirubin, haemoglobin, triglycerides, and fractionated heparin was evaluated. Results. Repeatability, intermediate imprecision, bias, and total error are overall below the defined limits of acceptability. Of interest is the high degree of agreement between Solea 100 and STA-R MAX with respect to PT (s), which fits perfectly with the theoretical line of identity (y = 0 + 1.00x). No interferences were found within the limits stated by the manufacturer, with some exceptions for APTT with heparin and APTT and PT for higher bilirubin concentrations. Conclusions. In conclusion, the performance of the Solea 100 optical analyser is satisfactory and adequate for the determination of routine coagulation tests. Moreover, they are perfectly comparable to mechanical systems, such as STA-R MAX and other upper-level analysers, even considering the low interference levels under routine conditions. Full article

(This article belongs to the Special Issue Diagnosis for Hematologic Disorders)

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9 pages, 1594 KiB

Open AccessArticle

Impact of Staining Methods and Human Factors on Accuracy of Manual Reticulocyte Enumeration

by Lin-Lin Pan, Hsiu-Chen Yu, Ching-Hui Lee, Kuo-Chuan Hung, I-Ting Tsai and Cheuk-Kwan Sun

Diagnostics 2022, 12(9), 2154; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics12092154 - 05 Sep 2022

Cited by 2 | Viewed by 4048

Abstract

Although peripheral blood reticulocyte enumeration reflects bone marrow functional integrity, which is important for differential diagnosis of hematological diseases, the factors affecting its accuracy have not been adequately addressed. Using 100 consecutive venous blood samples being processed with four supravital staining techniques [i.e., [...] Read more.

Although peripheral blood reticulocyte enumeration reflects bone marrow functional integrity, which is important for differential diagnosis of hematological diseases, the factors affecting its accuracy have not been adequately addressed. Using 100 consecutive venous blood samples being processed with four supravital staining techniques [i.e., brilliant cresyl blue (BCB), new methylene blue (NMB), and BCB/NMB with Liu’s stain] for reticulocyte enumeration, two technologists (senior vs. junior) conducted microscopic counting. The results were compared with those obtained with an automated system (Sysmex XE-5000) that served as the standard. The aims of this study were to identify (1) the technique that gave the most reliable outcome, and (2) possible human factors (i.e., seniority, repeated counting) that may affect the counting results. Analysis showed least bias (i.e., deviation from automated counting) associated with BCB staining, followed by NMB. In addition, the senior observer exhibited a higher bias in counting compared with their junior counterpart. Repeated counting also correlated with a higher rate of bias. Nevertheless, inter-observer consistency was high (intraclass correlation coefficient >0.95) and inter-/intra-observer variations were non-significant (both p > 0.05). Our results supported the use of BCB stain for reticulocyte enumeration and the reliability of manual counting despite the involvement of human factors, which had negligible impacts on the final outcomes. Full article

(This article belongs to the Special Issue Diagnosis for Hematologic Disorders)

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12 pages, 288 KiB

Open AccessArticle

Impact of Anti-Endothelial Cell Antibodies (AECAs) in Patients with Polycythemia Vera and Thrombosis

by Rossella Cacciola, Elio Gentilini Cacciola, Veronica Vecchio and Emma Cacciola

Diagnostics 2022, 12(5), 1077; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics12051077 - 25 Apr 2022

Cited by 3 | Viewed by 2045

Abstract

Polycythemia vera (PV) causes thrombosis. Erythrocytosis and cell adhesiveness are responsible for thrombosis. JAK2V617F causes inflammation and autoimmunity; however, whether or not autoimmunity or inflammation causes thrombosis has yet to be proven. In 60 PV patients, we analyzed JAK2V671F and its allele burden, [...] Read more.

Polycythemia vera (PV) causes thrombosis. Erythrocytosis and cell adhesiveness are responsible for thrombosis. JAK2V617F causes inflammation and autoimmunity; however, whether or not autoimmunity or inflammation causes thrombosis has yet to be proven. In 60 PV patients, we analyzed JAK2V671F and its allele burden, autoimmune Th17 cells, interleukin-17 (IL-17), anti-endothelial cell antibodies (AECAs), endothelial leukocyte adhesion molecule-1 (ELAM-1), intercellular adhesion molecule-1 (ICAM-1), and von Willebrand factor antigen (VWF: Ag). Fifty blood donors were used as the controls. All patients were on phlebotomy-maintaining hematocrit <45% and aspirin. Of the 60 patients, 40 had thrombosis. Those patients with thrombosis had a higher JAK2V617F allele burden than those without thrombosis, andTh17 cells and IL-17 were also higher in patients with thrombosis. Interestingly, we observed a high AECA IgG ELISA ratio (ER) in patients with thrombosis, which was normal in patients without thrombosis. We found high ELAM-1 and ICAM-1 as well as high VWF:Ag in patients with thrombosis compared to patients without thrombosis. AECA-positive sera from patients with thrombosis showed enhanced binding to cytokine-treated HUVEC and a positive antibody-dependent cellular cytotoxicity, suggesting that AECA may contribute to vascular injury. A positive correlation between AECAs, allele burden, and thrombosis was found. These results suggest that autoimmunity may be an additional mechanism in PV thrombogenesis. Full article

(This article belongs to the Special Issue Diagnosis for Hematologic Disorders)

14 pages, 712 KiB

Open AccessArticle

Detection of Unknown and Rare Pathogenic Variants in Antithrombin, Protein C and Protein S Deficiency Using High-Throughput Targeted Sequencing

by Petr Vrtel, Ludek Slavik, Radek Vodicka, Julia Stellmachova, Martin Prochazka, Jana Prochazkova, Jana Ulehlova, Peter Rohon, Tomas Simurda, Jan Stasko, Ivana Martinkova and Radek Vrtel

Diagnostics 2022, 12(5), 1060; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics12051060 - 23 Apr 2022

Cited by 4 | Viewed by 1867

Abstract

The deficiency of natural anticoagulants—antithrombin (AT), protein C (PC), and protein S (PS)—is a highly predisposing factor for thrombosis, which is still underdiagnosed at the genetic level. We aimed to establish and evaluate an optimal diagnostic approach based on a high-throughput sequencing platform [...] Read more.

The deficiency of natural anticoagulants—antithrombin (AT), protein C (PC), and protein S (PS)—is a highly predisposing factor for thrombosis, which is still underdiagnosed at the genetic level. We aimed to establish and evaluate an optimal diagnostic approach based on a high-throughput sequencing platform suitable for testing a small number of genes. A fast, flexible, and efficient method involving automated amplicon library preparation and target sequencing on the Ion Torrent platform was optimized. The cohort consisted of a group of 31 unrelated patients selected for sequencing due to repeatedly low levels of one of the anticoagulant proteins (11 AT-deficient, 13 PC-deficient, and 7 PS-deficient patients). The overall mutation detection rate was 67.7%, highest in PC deficiency (76.9%), and six variants were newly detected—SERPINC1 c.398A > T (p.Gln133Leu), PROC c.450C > A (p.Tyr150Ter), c.715G > C (p.Gly239Arg) and c.866C > G (p.Pro289Arg), and PROS1 c.1468delA (p.Ile490fs) and c.1931T > A (p.Ile644Asn). Our data are consistent with those of previous studies, which mostly used time-consuming Sanger sequencing for genotyping, and the indication criteria for molecular genetic testing were adapted to this process in the past. Our promising results allow for a wider application of the described methodology in clinical practice, which will enable a suitable expansion of the group of indicated patients to include individuals with severe clinical findings of thrombosis at a young age. Moreover, this approach is flexible and applicable to other oligogenic panels. Full article

(This article belongs to the Special Issue Diagnosis for Hematologic Disorders)

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Review

Jump to: Research, Other

22 pages, 1841 KiB

Open AccessReview

Digital Droplet PCR in Hematologic Malignancies: A New Useful Molecular Tool

by Sara Galimberti, Serena Balducci, Francesca Guerrini, Marzia Del Re and Rossella Cacciola

Diagnostics 2022, 12(6), 1305; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics12061305 - 24 May 2022

Cited by 18 | Viewed by 8383

Abstract

Digital droplet PCR (ddPCR) is a recent version of quantitative PCR (QT-PCR), useful for measuring gene expression, doing clonality assays and detecting hot spot mutations. In respect of QT-PCR, ddPCR is more sensitive, does not need any reference curve and can quantify one [...] Read more.

Digital droplet PCR (ddPCR) is a recent version of quantitative PCR (QT-PCR), useful for measuring gene expression, doing clonality assays and detecting hot spot mutations. In respect of QT-PCR, ddPCR is more sensitive, does not need any reference curve and can quantify one quarter of samples already defined as “positive but not quantifiable”. In the IgH and TCR clonality assessment, ddPCR recapitulates the allele-specific oligonucleotide PCR (ASO-PCR), being not adapt for detecting clonal evolution, that, on the contrary, does not represent a pitfall for the next generation sequencing (NGS) technique. Differently from NGS, ddPCR is not able to sequence the whole gene, but it is useful, cheaper, and less time-consuming when hot spot mutations are the targets, such as occurs with IDH1, IDH2, NPM1 in acute leukemias or T315I mutation in Philadelphia-positive leukemias or JAK2 in chronic myeloproliferative neoplasms. Further versions of ddPCR, that combine different primers/probes fluorescences and concentrations, allow measuring up to four targets in the same PCR reaction, sparing material, time, and money. ddPCR is also useful for quantitating BCR-ABL1 fusion gene, WT1 expression, donor chimerism, and minimal residual disease, so helping physicians to realize that “patient-tailored therapy” that is the aim of the modern hematology. Full article

(This article belongs to the Special Issue Diagnosis for Hematologic Disorders)

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Other

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5 pages, 1756 KiB

Open AccessInteresting Images

Primary Effusion Lymphoma: A Rare and Challenging Diagnosis for Recurrent Pleural Effusion

by Letícia Jacome Pereira, Sara Mohrbacher, Precil Diego Miranda de Menezes Neves, Flavia Fernandes Silva Zacchi, Ivan Ucella Dantas Medeiros, Victor Augusto Hamamoto Sato, Érico Souza Oliveira, Leonardo Victor Barbosa Pereira, Américo Lourenço Cuvello-Neto, Otávio Baiocchi and Pedro Renato Chocair

Diagnostics 2023, 13(3), 370; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics13030370 - 19 Jan 2023

Cited by 2 | Viewed by 2138

Abstract

Primary Effusion Lymphoma is an extremely rare and aggressive subtype of B-cell lymphoma, accounting for only <1% of all cases of this neoplasm. It has a unique clinical presentation because it has a predilection for appearing in body cavities, such as the pleural [...] Read more.

Primary Effusion Lymphoma is an extremely rare and aggressive subtype of B-cell lymphoma, accounting for only <1% of all cases of this neoplasm. It has a unique clinical presentation because it has a predilection for appearing in body cavities, such as the pleural space, pericardium and peritoneum. It mainly affects immunocompromised individuals and may also affect individuals in the Mediterranean region and in areas endemic for human herpesvirus 8 (HHV-8). Herein, we report the case of an 83-year-old immunocompetent male complaining of coughing, fever and progressive dyspnea for 3 days. His past medical history revealed a recurrent pleural effusion for the last three years, as well as losing weight and malaise. A subsequent investigation revealed a PEL diagnosis of the pleura. Full article

(This article belongs to the Special Issue Diagnosis for Hematologic Disorders)

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