Thalassemia is one of the most common monogenic disorders in southern China, with the Hakka population exhibiting a distinct genetic profile. This study evaluated whether next-generation sequencing (NGS)-based universal screening offers advantages over traditional cascade screening in this specific population. We enrolled 91,857 reproductive-age individuals in Ganzhou, a high-prevalence Hakka region, comparing 37,500 participants who underwent traditional screening (2019-2021) with 54,357 who received NGS-based screening. NGS-based screening detected thalassemia carriers at a rate 4.74 times higher than traditional screening (15.05% vs. 3.18%), particularly identifying silent α-thalassemia and β++-thalassemia variants that were substantially under-detected by the traditional approach. Crucially, while traditional screening identified 98.7% of β‑high‑severity couples, it detected only 71.4% of α‑high‑severity couples and missed 28.6% of those at risk for Hb Bart's hydrops fetalis, as well as 32.7% of couples at risk for severe non‑deletional Hb H disease. Traditional screening cost only USD 3.90 per person (versus USD 21.00 for NGS) and provided faster results (6.40 vs. 13.06 days). NGS-based universal screening provides a more accurate assessment of thalassemia burden and genotype diversity, offering critical advantages for prevention programs. Although traditional cascade screening is more cost-effective, its limited sensitivity for α-thalassemia variants leads to substantial underdetection. These findings underscore the need to tailor screening protocols to local epidemiological and genetic characteristics.

Hemotropic Mycoplasma spp., also known as hemoplasmas, are blood-borne pathogens that infect red blood cells and can cause hemolytic anemia in mammals, including dogs. Mycoplasma haemocanis (Mhc) and 'Candidatus Mycoplasma haematoparvum' (CMhp) are the two most common canine hemoplasma species. However, molecular epidemiological data and information on their genetic diversity remain scarce in Thailand. This study investigated hemoplasma PCR positivity in 472 client-owned dogs from Bangkok and Nakhon Pathom and 199 community dogs from Chiang Mai using a polymerase chain reaction (PCR) assay targeting the 16S rRNA gene. The 23S rRNA and RNase P RNA (rnpB) genes were additionally amplified from positive samples for multilocus characterization. Overall, 48 of 671 dogs were PCR-positive (7.2%; 95% confidence interval: 5.4%-9.3%). PCR positivity was higher in the community-dog/Chiang Mai cohort than in the client-owned cohorts; however, dog type and sampling location were fully confounded, and these comparisons should therefore be interpreted descriptively. Sequence analyses identified Mhc and CMhp. Sequence data suitable for analysis at both the 16S and 23S rRNA genes were obtained from 29 positive samples, and rnpB sequences were obtained from nine of these samples. To the best of our knowledge, this study provides the first 23S rRNA gene sequences of CMhp; no homologous CMhp 23S rRNA gene sequence was available in GenBank for direct comparison.

Chronic automated red blood cell exchange (RCE) is a cornerstone therapy for preventing sickle cell disease (SCD)-related complications, particularly stroke and recurrent vaso-occlusive crises. Current scheduling commonly relies on uniform empirical intervals of 3-6 weeks and does not account for substantial inter-patient variability in hemoglobin S (HbS) rebound kinetics, potentially resulting in undertreatment of rapid rebounders and unnecessary procedures in slower rebounders. We hypothesized that each patient's personalized rate of rise (ROR) of HbS could be used to generate an individualized, risk-adjusted RCE interval. We retrospectively analyzed adult patients with SCD receiving chronic outpatient RCE. The primary cohort comprised 18 patients with HbSS, while a single patient with HbSC was analyzed separately. HbS values from consecutive outpatient procedures were used to calculate an individual ROR (%HbS/day). A personalized scheduling formula was derived: t(interval) = S(risk) × (HbS(threshold) - HbS(post))/ROR(personal), where S(risk) represents an adjustable safety factor for patients at higher clinical risk. Individualized intervals differed markedly from uniform scheduling. Slow rebounders (< 0.5%/day) could potentially extend beyond 5 weeks, whereas rapid rebounders (> 1.0%/day) reached prespecified HbS thresholds within approximately 2-3 weeks. For most patients, the model recommended shorter intervals than those used, suggesting that under current scheduling many patients may exceed their HbS threshold between procedures. Because most patients would require shorter rather than longer intervals to maintain HbS below 30%, strict implementation of the model could increase program-level blood utilization; therefore, its principal value lies in precision and resource reallocation rather than guaranteed net savings. A personalized ROR-based scheduling framework integrates objective HbS kinetics with clinical risk stratification and may be most applicable to patients with stable, reproducible rebound kinetics. Prospective validation and integration into electronic health record systems are warranted to determine whether model-guided scheduling improves clinical outcomes.

Thalassemia, hemoglobinopathies, and glucose-6-phosphate dehydrogenase (G6PD) deficiency represent highly prevalent and genetically heterogeneous inherited hematologic disorders or diseases in Thailand. Conventional molecular approaches require multiple targeted assays to detect deletional and non-deletional variants across globin gene clusters and the G6PD gene, resulting in increased complexity, turnaround time, and cost. We developed and analytically validated a comprehensive long-read sequencing workflow using Oxford Nanopore Technologies for simultaneous detection of pathogenic variants in HBA 1, HBA 2, HBB, and G6PD. A total of 63 genomic DNA samples with previously characterized genotypes were analyzed using multiplex long-amplicon PCR followed by nanopore sequencing and bioinformatic variant calling with Clair 3. Variant annotation was performed using Ensembl VEP, and classification followed ACMG guidelines. All samples were successfully genotyped, demonstrating 100% concordance with conventional PCR, and Sanger sequencing. The platform accurately identified large α-globin deletions, β-thalassemia mutations, structural hemoglobin variants, and major Southeast Asian G6PD variants. A total of 39 distinct variants were detected and classified. Long-read sequencing effectively resolved homologous regions between HBA 1 and HBA 2 and enabled accurate characterization of multilocus co-inheritance patterns. This integrated nanopore-based workflow provides a robust, high-resolution, and scalable single-platform solution for comprehensive molecular diagnosis of thalassemia, hemoglobinopathies, and G6PD deficiency in genetically heterogeneous populations.

Red blood cell alloimmunization in pregnancy is a major cause of hemolytic disease of the fetus and neonate (HDFN), which can lead to fetal anemia, hydrops fetalis, and perinatal morbidity and mortality. The implementation of routine RhD prophylaxis has significantly reduced the incidence of RhD alloimmunization, especially in high-income countries. Management relies on maternal antibody screening, fetal antigen determination, and Doppler ultrasonography, with the middle cerebral artery peak systolic velocity being the gold standard for identifying fetuses at high risk of moderate-to-severe anemia that may benefit from intrauterine transfusion. Management depends on alloantibody type and titer, which usually correlate to the severity of fetal anemia. In cases of moderate-to-severe fetal anemia the primary treatment consists of intrauterine transfusion, which is an invasive procedure associated with an increased risk of fetal demise, particularly when performed prior to 20 weeks of gestation. Alternative therapeutic approaches, including intravenous immunoglobulin and plasmapheresis have been reported to delay or reduce the need for intrauterine transfusions, whilst the use of the neonatal Fc receptor inhibitor nipocalimab is currently under investigation. Despite significant advances, HDFN remains a global health concern, particularly in low-resource settings where access to RhD prophylaxis, non-invasive screening, and intrauterine transfusion is limited. A multidisciplinary management involving obstetrician, neonatologist, hematologist, and transfusion specialist is essential for optimizing outcomes. In this review, we provide an overview of the pathophysiology, diagnosis, management, and treatment options for red blood cell alloimmunization and HDFN.

Extramedullary haematopoiesis (EMH) is a rare cause of posterior mediastinal masses, often linked to chronic anaemia. A man in his late 60s with recurrent biliary disease was referred for evaluation of posterior mediastinal masses incidentally found during hospitalisation for choledocholithiasis. Imaging and biopsy revealed paravertebral EMH. Laboratory testing showed compensated haemolytic anaemia with stomatocytes, elevated eosin-5-maleimide binding, decreased erythrocyte osmotic fragility, and a PIEZO1 mutation (p.K2323T), leading to diagnosis of hereditary xerocytosis (HX). Family screening revealed the same condition in his son. HX, a rare congenital haemolytic anaemia, often remains undiagnosed due to mild symptoms and subtle findings. This case highlights HX as a unifying cause of recurrent biliary disease and posterior mediastinal EMH, emphasising the importance of recognising this rare condition to avoid inappropriate interventions such as splenectomy.