Objective:

• To discuss complex cases in immunhematology, focusing on the application of special reagents and methodologies to solve diagnostic and transfusion challenges.
• It presents two clinical cases that illustrate typical situations. The importance of techniques such as adsorption, elution and the use of specialized reagents (such as chloroquine and ZZAP) to characterize antibodies and guarantee safe transfusions, even in complex scenarios, is also emphasized.

Speaker: Regina Cardoso, MSc

Objective:

• To explore the variants of the RhD phenotype and how molecular alterations affect serological tests. It details types of mutations (deletions, insertions, gene conversions and single nucleotide variants), which can cause changes in RhD protein expression.
• These changes result in phenotypes such as D negative, Del, weak D and partial D, with different levels of antigenic expression and clinical impact. The importance of genetic and molecular understanding to improve diagnosis and safe transfusions is emphasized.

Speaker: Dr. Carine Prisco Arnoni

Objective:

• RhCE variants can complicate routine serology and require detailed investigation to determine the clinical significance of antibodies. Serological and molecular techniques are essential for the detection and management of RhCE variants.

Speaker: Dr. Tatiane Vendrame

Objective:

• The importance of molecular genotyping in clinical immunohematology, in diagnosis, transfusion management, and prevention of alloimmunization.
• It explores how genotyping techniques, such as next generation sequencing (NGS), overcome the limitations of serological tests, especially in cases of frequent transfusions.
• Practical examples demonstrate how genotyping improves transfusion compatibility and reduces complications, and the impact of genotyping on improving the safety and efficacy of transfusion medicine is emphasized.

Presenter: Lilian Castilho, PhD

Objective:

• Molecular techniques used to determine blood group alleles, emphasizing their application in immunohematology.
• Techniques such as PCR, PCR-RFLP, next generation sequencing and microarrays are highlighted for their efficiency in identifying clinically relevant alleles and rare variants.
• The importance of these tools to personalize and optimize transfusion management, promoting greater patient safety, is highlighted.

Presenter: Carine Prisco Arnoni

Objective:

• Antibodies of Undetermined Specificity (AUS) are antibodies or reactions that remain unexplained after excluding common antibodies, usually identified in irregular antibody screening (IAP) tests.
• Identification includes the use of extended phenotyping, different panels and chemical treatments (e.g. chloroquine, enzymes) to clarify specificities, as well as techniques such as adsorption/elution.

Presenter: Regina Cardoso , MSc

Objective:

• Solid phase platelet crossmatch has an application, methodology and clinical cases of Platelet Refractoriness or inadequate platelet increase after transfusions, usually associated with antibodies against platelet antigens (HLA, HPA).
• Detection techniques include the use of methods such as flow cytometry, PIFT, MAIPA and solid phase methodology (Capture-P), the latter used to detect antiplatelet antibodies and transfusion compatibility.
• Examples include strategies for refractory patients, the use of desensitization protocols and slow transfusions of incompatible platelets in critical cases.

Presenter: Juliana Pires Marafon Franz, MD, PhD

Isohemagglutinin tests (anti-A and anti-B) are antibodies that can be IgM, IgG or IgA, all of which have significant clinical potential and can cause intra- and extravascular hemolysis. ABO incompatibility (major or minor) can reduce the effectiveness of the transfusion and cause adverse reactions such as acute hemolysis. Practical examples demonstrate the seriousness of incompatible transfusions and the importance of prophylactic strategies. Example of local validation using automation to determine critical titers and define PC selection protocols. Presenter: Ana Paula Rocha Diniz Zanelli

Objective: This article discusses Platelet Crossmatch, an essential method for platelet compatibility in refractory patients. The technique is used to identify compatible platelets in alloimmunized patients. The method is based on the solid phase (Capture-P®), which detects IgG antibodies against platelets. It highlights the importance of standardized methods to improve the management of refractory patients and optimize clinical results. Speaker: Cristina Altobeli de Brito

Objective: To address RhD variants and the importance of their serological detection to guide molecular investigations. It explains how mutations such as SNPs, deletions and gene conversions result in variants such as partial D, weak D and Del. These alterations can cause differences in epitope expression and antisera reactivity. It includes examples of discrepancies in RhD typing and molecular characterization, such as a donor with a CDe haplotype and a patient with myelodysplastic syndrome. The importance of standardized methods and knowledge of variants for safe transfusion practices and accurate diagnoses is reinforced. Speaker: Dr. Tatiane de Paula Vendrame

Objective: A study of RhCE variants is presented, covering detection, classification and clinical significance. RhCE variants are detailed as variant proteins due to genetic polymorphisms, which can affect the expression of RhCE antigens such as C, c, E and e. Weakened expression, partial antigens and the absence of high-frequency antigens are mentioned. The need for greater knowledge of these variants to improve transfusions and blood compatibility in chronically transfused patients is emphasized. Speaker: Dr. Carine Prisco Arnoni

Objective: The use of molecular typing of blood group genes as a diagnostic tool and its application in transfusion medicine. It covers the characteristics of erythrocyte antigens, advances in genotyping and sequencing techniques, offering greater precision in the identification of rare variants and molecular compatibility, clinical cases and applications, highlighting the importance of personalized medicine in the transfusion context.

Speaker: Lilian Castilho, PhD

Objective: To review the impact of the use of the Microarray BeadChip (Immucor) in the identification of variants of the RHD and RHCE genes, highlighting its application in immunohematology and transfusion. The Microarray BeadChip as an efficient, sensitive and essential tool for solving complex immunohematology problems.

Speaker: Tamires Delfino dos Santos

Objective: To present the isohemagglutinin test, addressing its validation and impact on platelet concentrate transfusions. It includes a case of hemolytic reaction associated with incompatible transfusions, highlighting the importance of preventive measures. It presents data on critical titrations, methodologies and guidelines for minimizing risks in transfusions. Speaker: Ana Paula Rocha Diniz Zanelli

Objetivo: Revisar el uso de reactivos especiales en inmunohematología para resolver casos clínicos complejos como las discrepancias de grupo ABO y RhD, la presencia de anticuerpos irregulares y la anemia hemolítica autoinmune (AHAI), que requieren técnicas diagnósticas complementarias. Enumera las ventajas de los reactivos especiales para facilitar el trabajo de laboratorio, normalizar los procesos, mejorar la precisión diagnóstica y aumentar la seguridad transfusional.

Objetivos do seminário web:

• Isohemaglutininas e a Transfusao de Plaquetas
• validação do crossmatch plaquetario por fase sólida para manejo da Refratariedade Plaquetaria.
• inovações para a identificaçao de anticorpos de especificidade não determinada e as técnicas moleculares na determinação de alelos de grupos.

Orador: Jayanna Slayten, MS, MT(ASCP)SBBCM

Objetivos do seminário web:

• validação e conduta nas transfusões de plaquetas
• Crossmatching plaquetário
• Refratariedade Plaquetária.

Speakers: Ana Paula Rocha Diniz Zanelli; Cristina Altobeli; Juliana Pires Marafon Franz

Objetivos do seminário web:

• Reagentes Especiais em Imunohematologia: como usar e qual a importância na resolução de casos clínicos complexos
• A importância da detecção sorológica para direcionar a investigação molecular
• Tipagem molecular dos genes de grupos sanguíneos no auxílio diagnostico.

Speakers: Regina Cardoso; Tatiane de Paula Vendrame; Carine Arnoni, Tamires Delfino dos Santos, Lilian Castilho, PhD