Effect of Aboand Rh Blood Type on Sars-Cov-2 Infection Severity in Patients with Rheumatic Diseases: Data from the National Sar-Covid Registry

Objectives: To evaluate the association between the ABO and Rh antigens and the clinical characteristics and evolution of the SARS-CoV-2 infection in patients with rheumatic diseases. Method(s): SAR-COVID is a national, longitudinal, and observational registry. Patients >=18 years of age with a diagnosis of inflammatory or degenerative rheumatic disease, and confirmed SARS-CoV-2 infection (RT-PCR or serology) were included. Data were collected from August 2020 to June 2022. Sociodemographic, clinical data, comorbidities, underlying rheumatic disease, disease activity, and its treatment at the time of infection were recorded, aswell as symptoms, complications and treatments received for COVID-19. The WHO ordinal scale (WHO-OS) was used, and severe COVID-19was defined as WHO-OS>=5. Patients were categorized as follows: blood group A or non-A, and Rh factor positive or negative. Result(s): A total of 1356 patients were included, 547 (40,3%) had blood group A and 809 non-A (59,7%). Regarding the Rh factor, 1230 (90,7%)were positive and 126 (9,3%) negative. Age, sex, ethnicity and comorbidities were comparable between both groups. In both cases, the most frequent rheumatic diseases were rheumatoid arthritis (38,9%;p = 0,052), systemic lupus erythematosus (17,4%;p = 0,530) and osteoarthritis (10,1%;p = 0,888). Patients with non-A blood type presented a higher frequency of psoriatic arthritis (group A 5,1% vs non-A 8,7%;p = 0,015). During SARS-CoV-2 infection, more than 90% of patients in both groups were symptomatic (group A 96.0% vs non-A 94,8%;p = 0,384). Non-A blood group patients had a significantly higher frequency of arthralgia and dysgeusia. In A blood group 18.5% of the patients required hospitalization, 41,0% of them were admitted in the intensive care unit and 5.9% presented complications, while in the non-A blood group, were 16,7%, 31,1% and 5,5%, respectively (p > 0,05 in all the cases). The most frequent complications in both groups were respiratory distress syndrome and sepsis (p > 0,05). The outcome of the COVID-19 infection is detailed in Figure 1. In the multivariate analysis, adjusted for poor prognostic factors, patients with A blood type and those with negative Rh factor presented more likely severe COVID-19. (OR 1,75, 95%CI 1,20-2,56, p = 0,003 and OR 2,63, 95%CI 1,45-4,55, p = 0,001, respectively). Conclusion(s): Blood type A and negative Rh factor were associated with worse COVID-19 outcomes in this national cohort of patients with rheumatic diseases.

Molecular genetics and genomics of the ABO blood group system

The A and B oligosaccharide antigens of the ABO blood group system are produced from the common precursor, H substance, by enzymatic reactions catalyzed by A and B glycosyltransferases (AT and BT) encoded by functional A and B alleles at the ABO genetic locus, respectively. In 1990, my research team cloned human A, B, and O allelic cDNAs. We then demonstrated this central dogma of ABO and opened a new era of molecular genetics. We identified four amino acid substitutions between AT and BT and inactivating mutations in the O alleles, clarifying the allelic basis of ABO. We became the first to achieve successful ABO genotyping, discriminating between AA and AO genotypes and between BB and BO, which was impossible using immunohematological/serological methods. We also identified mutations in several subgroup alleles and also in the cis-AB and B(A) alleles that specify the expression of the A and B antigens by single alleles. Later, other scientists interested in the ABO system characterized many additional ABO alleles. However, the situation has changed drastically in the last decade, due to rapid advances in next-generation sequencing (NGS) technology, which has allowed the sequencing of several thousand genes and even the entire genome in individual experiments. Genome sequencing has revealed not only the exome but also transcription/translation regulatory elements. RNA sequencing determines which genes and spliced transcripts are expressed. Because more than 500,000 human genomes have been sequenced and deposited in sequence databases, bioinformaticians can retrieve and analyze this data without generating it. Now, in this era of genomics, we can harness the vast sequence information to unravel the molecular mechanisms responsible for important biological phenomena associated with the ABO polymorphism. Two examples are presented in this review: the delineation of the ABO gene evolution in a variety of species and the association of single nucleotide variant (SNV) sites in the ABO gene with diseases and biological parameters through genome-wide association studies (GWAS).Copyright © Annals of Blood. All rights reserved.

Mucormycosis in COVID-19 patients: A tertiary care experience

Introduction: Zygomycetes consisting of Mucorales order is a group of fungal infections. These species cause life threatening opportunistic fungal infections mucormycosis. This infection is highly prevalent in immunocompromised. During the 2 nd wave of Covid 19 pandemic corticosteroid treatment was used which has been linked to development of Mucormycosis. In our tertiary care teaching hospital we saw that patients suffering from Covid-19 infections developed mucormycosis. We present these cases in our study. To study the clinical, demographical,and Laboratory parameters in Covid-19 patients with Mucormycosis. Material(s) and Method(s): Retrospective Study. All biopsy proven cases of Mucormycosis (which developed after Covid-19 infection) were included. Relevant Clinical Demographics and Laboratory data was retrieved from the available case sheets. The data was tabulated in Excel sheet and further reviewed. Result(s): A total of 22 patients were diagnosed as suffering from mucormycosis majority were unvaccinated. 11 patients out of 22 (50%) started manifesting mucormycosis within one week of COVID infection. All the patients who had only single comorbidity (22.72%) suffered from mild disease and patient who had more than one comorbidity suffered from moderate (27.27%) to severe (50%) COVID infection. Conclusion(s): It is suggested that patients with Covid-19 infection are at risk for development of opportunistic fungal infections like Mucormycosis. Hence the physicians who are involved in treating such patients must be mindful of the fact that mucormycosis can develop in them. Histopathology helps in establishing a concrete diagnosis of Mucormycosis.Copyright © 2022 Wolters Kluwer Medknow Publications. All rights reserved.

SARS-CoV-2 and Helicobacter pylori and some hematological parameters: A case-control study

Background: The SARS-CoV-2 associated with bacterial infection represents a serious public health challenge. Recently, there is a remarkable increase in the number of researches that confirms the effect of Helicobacter pylori on pulmonary diseases. Aim(s): The goal of this research was to see how H. pylori affected the presentation of COVID-19 infections as a prospective risk factor. Material(s) and Method(s): This research was conducted in Babylon, Iraq, from January 1, 2022, to March 5, 2022. A total of 180 people were engaged in this study, with 90 patients identified with SARS-CoV-2 by polymerase chain reaction testing and 90 people serving as a control group. Antibody screening assays on blood samples were used to look for antibodies against H. pylori. The samples were processed for complete blood count and ABO blood group. Result(s): COVID-19 infection was more frequent in females than in males, especially between 31 and 45 years. When compared to healthy people, COVID-19 patients had a higher white blood cell count (P = 0.0001) and a lower lymphocyte count (P = 0.0001). H. pylori and COVID-19 have been found to have a strong relationship, especially in females. When comparing patients to healthy people, blood group A is the most common. Conclusion(s): People with H. pylori infections are considerably more sensitive to COVID-19 than people without H. pylori infections (P = 0.011). In combination with SARS-CoV-2, IgG for H. pylori might be a risk factor.Copyright © 2023 Journal of Medical Society Published by Wolters Kluwer-Medknow.

The Risk of COVID-19 in People Having a Particular Set of Gene

These risk factors of advancing age, male gender and co-existing health conditions like cancer, cardiovascular diseases, diabetes and obesity do not fully explain why some people have no or mild symptoms whereas others have severe symptoms. Genomewide association study (GWAS) identify a 3p21.31 gene cluster as a genetic susceptibility locus in patients with COVID-19 with respiratory failure. They also found a higher risk among persons with blood group A and protective effect for blood group O than among patients with other blood groups. The particular haplotype in a region of chromosome 3 is contributed to modern humans by neandertals. Another Neanderthal haplotype on chromosome 12 is associated with a 22% reduction in relative risk of becoming severely ill with COVID-19. The ApoE e4e4 homozygous genotype was found to increase the risk of severe COVID-19. Change in angiotensin converting enzyme (ACE) 2 gene was also found to be associated with increased risk of COVID-19, cardiovascular and pulmonary conditions. Copyright © 2021, Kathmandu University. All rights reserved.

Probability of Covid-19 in Abo Blood Type during Second Wave in Southern Rajasthan, India

Objectives: Along the course human history of scientific research, the severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) is the most concerning global health problem. Second wave of COVID-19 has adversely affected India. However, India embarked on its immunization program on January 16, 2021, operating 3006 vaccination centers onset Covaxin and CoviShield. This study aimed to ascertain if there is an association amidst ABO blood type and probability of COVID-19 infection in wave. Method(s): This is analytical and observational study conducted on 713 SARS-COVID-19-positive patients of a known ABO blood type, who attended outpatient department and inpatient department during March 26-May 20, 2021, in tertiary care hospital Udaipur (Raj.) Serum inflammatory markers were evaluated by Cobas 6000. Result(s): Out of the 713 patients who were tested positive, 15.56% was blood group Type A, 19.91% was blood group Type B, 13.65% was blood group Type AB, and 46.28% was blood group Type O. On statistical analysis, there were positive association between O+ blood type and peak inflammatory marker (interleukin-6 and D-Dimer). Patients with blood Type O who received a test were more likely to test positive and blood Type B+, A+, A+, AB+, O-, A-, B-, and AB- were less likely to test positive. Conclusion(s): The present study shows an evidence for interrelation between ABO blood groups and SARS-COVID-19. Reported infection prevalence is moderately increased among O+ blood type individuals. Determination of level of inflammatory markers might prove to be helpful to clinicians so as to keep track of severity of infection and evaluate the prognosis of SARS-COVID-19 with specific ABO blood groups. Copyright © 2022 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)

A Influencia Do Sistema De Grupo Sanguineo Abo Na Suscetibilidade E Letalidade Da Covid-19

Objetivo: Realizar um estudo descritivo, sintetizando, analisando e discutindo as informacoes mais recentes sobre a influencia do sistema de grupo sanguineo ABO na COVID-19, a partir de uma revisao sobre a biossintese e base molecular do sistema ABO e sua relacao com a suscetibilidade a infeccao pelo SARS-CoV-2 e letalidade da doenca, a partir da sua relacao com trombofilias e eventos cardiovasculares na COVID-19. Metodologia: Foram realizadas pesquisas em bases de dados e bibliotecas cientificas, tais como Pubmed, Scielo, e Nature, utilizando palavras-chave como "COVID-19", "SARS-CoV2", "Coronavirus", "Sistema de Grupo Sanguineo ABO", "Hemostasia", "Trombose", "Fator de von Willebrand", "ADAMTS13" e, ao todo, foram utilizados 53 trabalhos, entre livros e artigos cientificos, dos quais as informacoes utilizadas foram cuidadosamente selecionadas a fim atender ao objetivo proposto. Resultados/Discussao: Alem de sua elevada relevancia para a clinica transfusional, o sistema de grupo sanguineo ABO ja foi associado a suscetibilidade e fisiopatologia de diversas infeccoes e enfermidades. Com a pandemia do novo Coronavirus nao foi diferente. Estudos relacionaram este grupo de antigenos eritrocitarios a suscetibilidade e letalidade da COVID-19, e apontaram uma maior predisposicao a infeccao para o grupo A, bem como um pior prognostico da doenca, ao passo que, o reduzido numero de infectados e obitos no grupo O poderia indicar um efeito protetor nestes individuos. O fenotipo A esta relacionado a uma maior predisposicao a eventos tromboticos devido ao aumento do FVW na circulacao desses individuos, fator que, quando somado ao potencial trombogenico da doenca causada pelo SARS-CoV2, pode ser um agravante no seu prognostico, de modo a possibilitar a evolucao a um quadro mais grave com implicacoes cardiovasculares, que podem levar a sindrome do desconforto respiratorio agudo, alem da ocorrencia de trombose venosa profunda, acidente vascular encefalico isquemico, infarto agudo do miocardio, tromboembolismo pulmonar e trombose placentaria. Conclusao: E observada nao apenas uma maior vulnerabilidade em relacao ao grupo A na infeccao pelo SARS-CoV2, como tambem um maior risco de evolucao para a forma grave da doenca, possivelmente o que se deve a N-Acetilgalactosamina como elemento facilitador na ancoragem do virus as celulas suscetiveis. O efeito contrario e observado para o grupo O, que se encontra em menor proporcao nos numeros de infectados e de obitos pela doenca, sugerindo um efeito protetor nestes individuos, que poderia ser explicado pela presenca de anticorpos anti-A circulantes no soro, principalmente do isotipo IgG, que seriam mais efetivos na neutralizacao do virus contendo antigenos ABO em seu envelope. A predisposicao a eventos vaso-oclusivos a partir do Sistema de Grupo Sanguineo ABO em individuos nao-O, principalmente no grupo A, e um importante fator que, quando somado ao potencial trombogenico da COVID-19, pode ser um agravante significativo na letalidade da doenca causada pelo SARS-CoV2 nesses pacientes, uma vez que, apos infectados, a probabilidade de evolucao a um quadro mais grave com implicacoes tromboembolicas e mais elevada devido a maior concentracao plasmatica de FvW nesses individuos. Copyright © 2022

Correlacao Entre Os Grupos Sanguineos E as Alteracoes Imuno-Hematologicas Na Covid-19

Objetivo: Identificar padroes entre os grupos sanguineos e as alteracoes imuno-hematologicas em pacientes com COVID-19 atendidos no Hospital das Clinicas da Faculdade de Medicina de Botucatu (HCFMB), no periodo marco de 2020 a outubro de 2021. Material e metodos: Avaliou-se 186 pacientes internados com o teste RTq-PCR positivo para COVID-19 e esses foras separados em dois grupos: 105 pacientes nao transfundidos e 81 pacientes transfundidos. Selecionou-se os resultados dos exames de tipagem ABO e RhD, pesquisa de anticorpos irregulares (PAI), marcadores de hemolise (hematocrito, hemoglobina e bilirrubina), desidrogenase lactica (DHL) e hemocomponentes transfundidos. Resultados: No que tange a tipagem sanguinea ABO e RhD, em pacientes nao transfundidos, foi observado que 39 eram do tipo A (37%), 19 do tipo B (18%), 4 do tipo AB (4%) e 43 do tipo O (41%). Desses, 93 pacientes eram RhD positivos (89%) e 12 eram RhD negativos (11%). Ja para os pacientes transfundidos, 35 pacientes eram do tipo A (43%), 13 do tipo B (16%), 4 do tipo AB (5%) e 29 do tipo O (36%). Em relacao ao fator RhD, 69 pacientes eram positivos (85%) e 12 negativos (15%). Os resultados obtidos foram comparados com a frequencia da tipagem sanguinea presente na populacao brasileira, sendo observado que houve menos pacientes do tipo O infectados pelo novo Coronavirus (p = 0,0071), como tambem para transfusoes sanguineas realizadas (p = 0,0235). Os pacientes transfundidos apresentaram maior frequencia estatisticamente significativa (p < 0,0001) de anticorpos irregulares quando comparados com os pacientes nao transfundidos, com destaque para presenca de crioaglutinina. Observou-se diferenca estatistica significativa de alteracoes dos valores dos marcadores de hemolise nos pacientes transfundidos, em que as medias dos valores encontrados foram de 28,93% para hematocrito, 9,41g/dL para hemoglobina e 1,4mg/dL para bilirrubina, indicando a existencia de anemia e destruicao celular. Discussao: Em relacao a tipagem sanguinea ABO, observou-se que houve mais pacientes do tipo A transfundidos quando comparados com as outras tipagens, enquanto mais pacientes com tipagem O nao necessitaram de transfusoes sanguineas. Para ambos houve o predominio do fator Rh positivo. Segundo a literatura, o grupo sanguineo A foi associado a um risco aumentado de infeccao em relacao ao grupo O, devido a presenca de anticorpos naturais anti-A, os quais sao capazes de inibir de forma especifica a proteina Spike do virus de se ligar efetivamente aos receptores da ECA2. Na identificacao de anticorpos irregulares, as crioaglutininas estiveram presentes na maioria dos pacientes com PAI positivo e sua presenca prejudicou as analises laboratoriais, atrasando a liberacao dos resultados dos testes imuno-hematologicos. Conclusao: Portanto, identificou-se correlacao entre os resultados alterados dos marcadores de hemolise com o desenvolvimento da forma mais grave da COVID-19 e a necessidade de transfusao de hemocomponentes, sendo o concentrado de hemacias (CH) o mais utilizado para tratar a anemia estabelecida nesses pacientes. Copyright © 2022

Investigation of the clinical course of Covid-19 patients according to blood groups

The aim of this study is to evaluate the effect of blood groups (BGs) on Covid-19 contraction and prognosis and to reveal the coefficients. Patients who referred to Covid-19 outpatient clinics and had an established diagnosis of Covid-19 were included in the study. Their BGs, previous diagnoses and blood examination findings were retrospectively analyzed. Duration of hospitalization, clinical course and survival were recorded. The mean age of 365 subjects, 210 female and 155 male, was 45,5 years. Subjects with BG A developed Covid-19 at significantly higher rates (p = 0.001), while BG O was found associated with lower rates (p = 0.005). Lymphocyte count was found lower (p = 0.035) and rate of lung parenchymal involvement was higher (p = 0.003) in patients with Rh antigen. It was found that a higher percentage of patients with B BG required treatment in the intensive care unit (ICU) compared to other ABO BGs (p = 0.015). These results suggest a higher risk of Covid-19 contraction in the population with BG A and lower risk for BG O population while indicating poorer prognosis for patients with BG B. Copyright © 2022 Ondokuz Mayis Universitesi. All rights reserved.

ABO blood group as an additional risk factor for venous thromboembolism

Background: Higher thrombotic risk associated with non-O blood group individuals is observed especially when in combination with thrombophilia. Aim(s): To estimate the association of ABO blood type with venous thromboembolism (VTE). Method(s): In this retrospective case control study we enrolled 40 patients with confirmed VTE. In all of them ABO blood group was determined using serologic methods such as column agglutination technique which included forward and reverse typing. Thrombophilia testing was performed in 12 patients with unprovoked VTE event. Result(s): ABO distribution among VTE patients is as follows: A in 19 (47.5%), O in 5 (12.5%), B in 10 (25.0%) and AB in 6 (15.0%) patients. The prevalence of O blood group in VTE patients was significantly lower (12.5%) in comparison to non-O blood types (87.5%). Blood group A is more prevalent and group O is less prevalent in VTE patients in comparison to our general population in which the frequency of A and O blood group is 41% and 32% respectively. In 6 VTE patients Covid 19 was confirmed few weeks prior to the thrombotic event (4-A, 1-O and 1-B blood group). In 12 patients tested for thrombophilia, 36 mutations were discovered with the following frequency: FII G20210A-5.5%, FVL G1691A-13.8%, FXIII V34L-8.3%, FGB G455A-11.1%, ITGA2 C807T-5.5%, ITG B3 T1565C-2.7%, PAI-1 4G/5G675-27.7%, MTHFR C677T-25%, from which 24 were heterozygous and 12 were homozygous (2-FVL, 2-FXIII, 1-FGB, 6-PAI- 1 and 1 case with MTHFR mutation). All of the patients have more than one mutation in different combinations and their ABO distribution was similar as in the rest of the VTE patients. Conclusion(s): According to the results, individuals with non-O blood group are more prone to VTE. ABO phenotype and especially ABO genotype may be additional diagnostic tool for assessment of the VTE risk together with thrombophilia markers.

COVID-19+ Kidney Transplant in US Veterans: An Opportunity to Increase Transplant Rates

Purpose: US veterans are less likely to be transplanted. This disparity is related to donor shortage. This study seeks to increase access to transplant of US veterans to transplants using covid+ organs. This provides high quality organs to underserved population, reduce discards and is safe. Report of safety of this strategy. Early reports demonstrates safety in using these organs. Method(s): Case presentation of two veterans transplanted with covid+ organs. Recipient and donor characteristics highlighted including vaccine status, antibody level (IgG) against Sars-CoV-2 prior to and following transplant. Organ function is reported and complications. Induction agent using non-depleting agent recorded. Informed consent obtained for covid+ organ. Infectious disease consultation. IgG antibody level check prior to transplant, and post transplant. Result(s): Two cases reported. Both had no complications related to covid. Excellent donors with KDPI <30. Cycle threshold 16 and above. Sequence 193 and 225. EPTS >85. Simulect and steroids used in both cases. Cold time short >16 hours. Both brain dead donors. LOS 7 and 8 days. PRA 0%. Donor blood group A and O. DGF in one recipient which recovered after 2 weeks see figure. The recipients had initially high antibody level which declined post transplant by 50%. See fig. Conclusion(s): Using Sars-CoV-2 donors is safe and provides a source of potential high quality organs in US Veterans who are generally underserved using informed consent. Full vaccination in recipient is a prerequisite. (Table Presented).

BLOOD GROUPS AND ACUTE RESPIRATORY VIRAL INFECTION COVID-19

Introduction. Polysaccharides, glycoproteins and glycolipids, which determine the group-specific properties of human blood, are both structural elements of the whole organism and determine its predisposition to certain somatic and infectious diseases. Thus, the blood group of an individual can be used among other markers and/or prognostic factors of the occurrence and course of certain groups of diseases. Aim - analysis of literature sources characterizing the relationship of blood groups with COVID-19 ARVI, as well as the mechanisms underlying this relationship. Main findings. The Oaß(I) phenotype ensures an individual's resistance to infection with the SARS-CoV-2 virus and allows for a relatively mild course of the disease. The Aß(II) phenotype is a risk factor for the development of COVID-19 ARVI, in its severe course, the occurrence of complications and increased mortality. An additional component of protection in the form of a negative Rh-affiliation of the infected person is not excluded. The protective properties of the Oaß(I) phenotype are associated with the absence of polysaccharide A in an individual and the presence of anti-A antibodies. The increased risk of COVID-19 ARVI among Aß(II) individuals is due to the large polymorphism of polysaccharide A in the environment and the lack of natural immunity to other forms of polysaccharide A in this group.

Case report: Anti-p1 and AB blood group-A faux Para- Bombay

Background: Anti-P1 is a common antibody found in the sera of P2 donors, affecting one-quarter to two-thirds of those tested. Anti-P1 is an IgM isotype antibody that is frequently found as a weak cold agglutinin. Anti-P1 antibodies that are reactive at 37 Celsius or cause in vitro hemolysis are rare. With the exception of the rare Bombay phenotype, all red cells express the H antigen. The amount of H antigen on red cells is determined by an individual's ABO type since H antigen is the precursor to both A and B antigens. The expression of the H antigen is highest in group O and lowest in group A1B (O>A2 > B > A2B > A1 > A1B). We report a case of blood discrepancy mimicking Para-Bombay due to anti-P1 and weak H antigen expression in a 46-year-old Sarawak Malay blood donor during routine blood donor regrouping with an automated immunohematology analyser. She has history of COVID19 infection in September 2021 and she completed her 1st, 2nd and booster mRNA vaccine in November 2021. Her last pregnancy was 13 years ago, and she has no history of blood transfusions. Aims: To resolve blood group discrepancies detected when using an automated immunohematology analyser. To understand the possibility of interference from natural occurring cold-reacting red cell alloantibodies during indirect antiglobulin test blood grouping. To understand the possibility of false negative in forward grouping with anti-H antisera in donors with A1B blood group. Methods: Blood donor was typed for ABO and Rh by an automated immunohematology analyser with microplates. Serological methods for antibody detection and specification were done manually with column agglutination method (gel-card) and tube method. Results: Forward grouping of the donor's first sample with an automated analyser was strongly positive for Anti-A (4+), Anti-B (4+), Anti-AB (4+) and Anti-D (4+), while reverse grouping was also strongly positive for A1-cell (3+), B-cell (3+) and O-cell (4+). Manual serological methods with gel-card and tube method yielded similar results. Anti-H showed no reaction. The first sample was negative for Direct Coomb's test (DCT). The donor's second (repeat) sample using the manual serological method yielded similar results;however, reverse grouping repeated at 37 Celcius resulted in the cessation of reactions on known cells. Anti-H showed a 1+ reaction. Antibody screening was positive and proceeded to 11 panel antibody identification with Anti-P1 identified. DCT was negative in the second sample. (P1-) and Le(a-b+) are her phenotypes. Summary/Conclusions: Anti-P1 is commonly reported as cold reacting alloantibody in patients. In this case, a combination of strong reacting anti-P1 at room temperature and commonly low H antigen volume in A1B red cells lead to a false initial suggestion of Para- Bombay phenotype. Blood grouping discrepancies detected with automation should always be repeated manually.

Collection of COVID-19 convalescent plasma in institute for transfusion medicine of republic of North Macedonia and evaluation of anti-SARS-CoV-2 antibody concentration in different donors subgroups

Background: Plasma collected from patients that have recovered from an infectious disease has been transfused over many decades for prophylaxis and treatment of various infectious diseases. Taking into consideration the expansion of COVID-19 pandemics, we started the COVID-19 convalescent plasma (CCP) programme. Aims: The aim of our study is to show our experience with collecting the CCP and to evaluate the SARS-CoV-2 antibody concentration in different convalescent plasma donors' subgroups. Methods: This is a prospective study performed in the Institute for Transfusion Medicine of Republic of North Macedonia since 30 April 2020 till July 2021. Antibody testing was performed at the Institute for Immunobiology and Human Genetics in Skopje using CLIA method with Snibe Maglumi SARS-CoV-2 S-RBD IgG (quantitative) with IgG cut-off larger than 5 AU/ml. All potential donor were tested for: negative RTPCR for SARSCoV-2 before donation, anti-SARS-CoV-2 antibodies, anti- HLA antibodies (where applicable), blood count, blood group, TTI and biochemistry. Preferred method for plasma collection was plasmapheresis which was performed with Terumo BCT Trima Accel and donation of whole blood, depending on the donor preference and venous access. All donors signed inform consent for donation and inclusion in the study. Results: There were 1476 potential CCP donors, but only 700(47.9%) donors fulfilled all the criteria and we obtained 793 units of CCP;639 (80.6%) units from whole blood donors and 154 (19.4%) CCP units from 61 plasmapheresis donors, 485 (69.3%) males and 215 (30.7%) females. Mean age of the donors was 40 years (range 18-63). Mean value of SARS-CoV-2 S-RBD IgG concentration was 31.05 AU/ml, (range from 5.1 AU/ml to >100 AU/ml), mean value of SARS-CoV-2 S-RBD IgG in men was 37.6 AU/ml and 28.9 AU/ml in women (p < 0.05). Distribution of CCP donors according to the ABO blood group was: 301 blood group A (43%) with median value of SARS-CoV-2 S-RBD IgG = 27.15 AU/ml, 220 blood group O (31.4%) median value of SARS-CoV-2 S-RBD IgG = 32.1 AU/ml, 116 blood group B (16.6%) median value of SARS-CoV-2 S-RBD IgG = 35.9 AU/ml and 63 donors had blood group AB (9%) median value of SARS-CoV-2 S-RBD IgG = 26.45 AU/ml. There were 69 donors that were previously hospitalized with mean value of SARS-CoV-2 S-RBD IgG = 48.6 AU/ml, and 629 that were treated at home with mean value of SARS-CoV-2 SRBD IgG = 29.1 AU/ml (p < 0.05), of which 578 had symptoms with mean value of SARSCoV- 2 S-RBD IgG = 29.1 AU/ml and 51 were asymptomatic with mean value of SARS-CoV-2 S-RBD IgG = 29.3 AU/ml. The CCP donors had the following distribution according to the age: 125 donors in the 18-29 age group with median value of SARS-CoV-2 S-RBD IgG = 23.0 AU/ml, 200 donors in the 30-39 age group with mean value of SARSCoV-2 S-RBD IgG = 28.2 AU/ml, 217 donors in the 40-49 age group with mean value of SARS-CoV-2 SRBD IgG = 32.9 AU/ml and 156 donors in the 50-63 age group mean value of SARS-CoV-2 S-RBD IgG = 38.3 AU/ml (p < 0.05). Summary/Conclusions: The collection procedures are safe and effective and collected CCP units were with high concentration and quality. The concentration of SARS-CoV-2 S-RBD IgG in CCP obtained from previously hospitalized patients was significantly larger than in ones that were treated at home. The concentration of SARS-CoV-2 S-RBD IgG was higher in men, in advanced age group and in donors with blood group B. The further studies are needed to clarify the impact of different variables on antibodies concentration/ titre in donors.

The spectrum of ABO hemolytic disease of the fetus and newborn in neonates born to Group O mothers

Background: ABO hemolytic disease of the fetus and newborn (ABOHFDN) is a frequent event, and usually a problem of the neonate rather than the fetus, however, it is difficult to predict the disease severity. Thus, there is a need to increase awareness towards ABOHFDN for optimizing care in terms of early diagnosis and adequate monitoring. Aims: To determine the frequency of ABO-incompatibility in neonates born to group O mothers and to assess the severity of ABO-HDFN in neonates and determine the neonatal outcomes. Methods: This prospective observational study was carried out from February 2020 to May 2021 after obtaining a written informed consent from the mothers. A total of 260 neonates born to blood group O mothers were recruited. The maternal red cell antibody screen (ABS) using a 3-cell panel (Diacell, Bio-Rad, Switzerland) and the neonatal direct antiglobulin test (DAT) were done by column agglutination technique (CAT). For DAT positive samples, the IgG subclass of anti-A/anti-B was determined using DAT IgG1/IgG3 screening cards (Bio-Rad, Switzerland) and a heat elution at 56°C was also performed. The maternal anti-A/anti-B IgG titers was determined by tube technique after treating the serum sample with 0.01 M di-thiothreitol (DTT). The neonatal total serum bilirubin (TSB) and other relevant parameters were also recorded. The requirement for treatment in terms of phototherapy and/or exchange transfusion (ET) and the neonatal outcome were also recorded. Due to travel restrictions during the ongoing COVID-19 pandemic, the follow-up was performed telephonically with parents 6-8 weeks after discharge. Results: Of the 260 group O mothers, none had positive ABS. Of the 260 neonates born to them as an outcome of singleton pregnancies, 84 with blood group O were excluded from the study. The overall frequency of ABO-incompatibility between mother and neonates was 67.69% (176/260). Out of 176 neonates, 77 (43.8%) were group A and 99 (56.2%) were group B, and 15 (8.5%) of them had a positive DAT. Overall, 26.7% (47/176) neonates received phototherapy and 172 (97.7%) neonates survived. The mean (±SD) duration of phototherapy (hours) was 34.17 (±25.67) hours and it ranged from 12- 120 h. Only 1 neonate required ET. None of the neonates required readmission. The median maternal IgG anti-A titre was 16 (8-64) (range: 2-512), while the IgG anti-B titre was 32 (32-64) (range: 4- 512) (p = <0.001). The maximal TSB in neonates had a significant positive association with neonatal birth weight (p = 0.045), maturity at birth (p = 0.037), positive DAT (p = 0.006) and requirement of phototherapy (p = <0.001). Neonatal DAT positivity was significantly associated with maternal IgG titers (p = <0.001), neonatal PCV (p = 0.017), maximal TSB (p = 0.006), requirement (p = <0.001) and duration of phototherapy (p = 0.024). At a cut-off of maternal IgG titre ≥64, it predicted the requirement of phototherapy with a sensitivity of 72.3% and a specificity of 72%. The relative risk (95% CI) of a DAT positive neonate requiring phototherapy was calculated to be 4.55 (3.12-6.33). Summary/Conclusions: The frequency of ABO-incompatibility in neonates born to group O mothers was 67.69% (176/260). The maternal IgG titre of anti-A/anti-B of 64 or more could be a good predictor for identifying the neonates at-risk for developing hyperbilirubinemia requiring further management and combining it with neonatal DAT further enhances the sensitivity to identify such at-risk neonates.

Transfusion errors in the COVID-19 ward resulting in changes in standard operating procedures (SOP)

Background: The pandemic of COVID-19 has led to alterations in SOP across the transfusion process, including administration of blood in COVID-19 wards. COVID-19 patients who present with symptomatic anaemia and have multiple risk factors will need blood transfusions. ABO-incompatible blood transfusions leading to acute haemolytic transfusion reaction is a rare but potentially fatal complication. The National Haemovigilance Coordinating Centre of the National Blood Centre, Malaysia reported the national incidence of incorrect blood components transfused (IBCT) in relation to total blood products transfused in 2019 to be 75 per 10,000 units. Five IBCTs reported were related to administration errors. Aims: We reported two IBCTs involving two patients who required blood transfusions in a COVID-19 ward. Patient 1 was a 74 year old man who complained of chest pain, with a haemoglobin (Hb) of 5.7 g/ dl. Patient 2 was a 50 year old woman with a Hb of 6.4 g/dl. When the two units of blood arrived on the ward, one doctor completed the pre-administration checklist in the 'clean' zone, which Nurse 1 then counter-checked. Nurse 1 put the two blood bags into separate transparent plastic bags and labelled them with the wrong patient's identity sticker. She then handed both blood bags to Nurse 2 in the 'dirty' zone, without the patients' blood compatibility labels, blood request forms and bedside checklist forms. Positive patient identification was not done by Nurse 2 and the transfusions commenced. Patient 1 complained of chills around 10 min into the transfusion. The error was only realized 35 min into the transfusion when the symptoms persisted and the temperature taken was 38°C. Patient 2, who was given a unit of group O blood that belonged to Patient 1, did not report any adverse reactions. Our aim is to identify the root causes of these IBCTs and to execute the necessary changes in pre-transfusion SOP to minimize future recurrences. Methods: Samples from both patients were investigated for transfusion reactions. Rechecking of blood groups was done manually with the test tube method. Direct and indirect anti human globulin tests (DAT/IAT) and recheck of cross-matching were performed with the column agglutination technology (CAT) method at 37/AHG phase. Urine samples were tested using urine dipsticks. Isohaemagglutinin (anti-A/B titre) was performed using the CAT method at 37/AHG phase. Plasma Hb was measured with a photometer and a microcuvette. Results: Both patient 1's post-transfusion samples (immediate and post-24 h) were O-RhD positive. Blood group of the donor's bag was B-RhD positive. DAT for both samples was positive with IgG(3+) and C3D (1+). IAT for both samples was negative. Recheck of crossmatching with both samples was incompatible (4+). Urine tests were negative for haemoglobin. A low anti-B titre of 1:16 was detected. Plasma Hb was measured twice at a low level below the reference range. Patient 2's workup was unremarkable. Summary/Conclusions: Two IBCTs occurred in the COVID-19 ward, with one major ABO-mismatched IBCT due to human errors and deviation from standard SOP. Pre-transfusion SOP was still unclear in the COVID-19 ward setting prior to these incidences. All medical personnel in the COVID-19 ward underwent retraining on safe transfusion practices. One COVID-19 patient's blood compatibility label, blood request form, bedside checklist form and blood bag should be brought into 'dirty' zone to be checked by two medical personnel at one time.

Relationship between the abo blood group and the coronavirus disease 2019 (COVID-19) severity: a population-based cohort study in the Valencian community (Spain)

Background: It has already been published the existence of an association between ABO blood groups and COVID-19 severity. These studies have reported that the ABO blood group is statistically associated with the acquisition of SARS-CoV-2 and survival after the virus infection. Researchers have found that blood group A was associated with a higher risk for COVID-19 infection and severity compared with others blood groups, whereas blood group O was associated with a significantly lower risk compared with non-O blood groups. Aims: This study aimed to demonstrate, in a population-based cohort study in the Valencian Community (Spain), the relationship between the ABO blood group and COVID-19 severity. Methods: A total of 567 regular blood donors from the Valencian Community (Spain) who had been infected by SARS-CoV-2 were recruited. They were divided into three different cohorts based on their symptoms and their requirements during the infection: asymptomatic donors, mild donors, and severe donors. Demographic data were also included. Asymptomatic donors did not have any symptoms;mild donors had the most common symptoms associated with COVID-19 without the need for hospitalization and severe donors needed hospitalization due to severity. For the statistical analysis, we compared the three cohorts with regular blood donors who donated blood in the last 3 years (256,203 donors) in the Valencian Community. Data were analysed using Chi-Square tests. p-values <0.05 were considered significant, standardized residuals were calculated to assess the contribution of each cell to the significance of the test. GraphPad Prism v.7.0 software was used. Results: Our results show that 63.33% of blood donors who had passed COVID-19 disease and required hospitalization (severe donors) were from blood group A while only 26.67% were from blood group O (p-value = 0.0002). The same results were observed in the established cohort for mild donors, with 47.01% and 39.08% from blood groups A and O (p-value<0.0001), respectively (Table 1). These data suggest that blood group A could be a risk factor for COVID-19 infection and group. Regarding asymptomatic donors, only 38.52% were donors from blood group A while 13.12% were donors from blood group B (Table 1). This data indicate that blood group B could be protective against COVID-19 severity (pvalue< 0.0001). Summary/Conclusions: Blood group A and blood group O donors showed a significantly higher and lower risk (p-value = 0.0002), respectively, to require hospitalization after the infection of COVID- 19. However, belonging to blood group B seemed to be associated with low-risk SARS-CoV-2 infection (p-value<0.0001).

Reduced levels of ABO anti-A and anti-B of IGM, IGG and IGA isotypes in the serum but not the saliva of individuals recovered from COVID-19

Background: Recent studies reported that individuals with ABO blood type O are underrepresented among patients infected with severe acute respiratory syndrome coronavirus SARS-CoV-2 compared with controls. Our preceding study results indicated a lower proportion of individuals with blood type O accompanied by a higher incidence of blood type AB in patients hospitalized with COVID-19 than in healthy blood donors. Thus, we hypothesized that the variable susceptibility to infection with SARS-CoV-2 might be related to interference caused by circulating ABO antibodies and further may be influenced by the antibody titers which vary widely between individuals. Aims: Therefore, we aimed to investigate ABO antibody levels, including IgM, IgG and IgA subclasses, in the serum and saliva of Caucasians (n = 187), who recovered from mild COVID-19 and to compare them with those of individuals who had never been infected with the virus. Further, a possible association between ABO antibodies and virusspecific total antibody concentrations in the COVID-19 convalescents as well as a potential relationship between the total IgA secreted in saliva and anti-A/anti-B specific IgA in saliva specimens were addressed. Methods: The convalescent study participants were recruited between June 2020 and February 2021. Individuals who had been hospitalized with COVID-19 or who were pregnant were excluded from the study. Two samples were collected within 2 months after the diagnosis (median days: 44) and approximately 2 months later (median days: 66 days). Isotype specific anti-A and anti-B were determined by flow cytometry on a FACS Canto II. The results were compared with the levels in samples from blood and saliva donors. The antibodies specific to SARS-CoV-2 as well as total IgA in saliva were tested by ELISA. Results: COVID-19 convalescents had significantly lower levels of anti-A and anti-B IgM, IgG and IgA in their serum than control subjects (p < 0.001). ABO antibody levels tested in saliva of participants who previously suffered from COVID-19 did not differ significantly from antibody levels tested in saliva controls (p ≥ 0.338). ABO antibody levels remained stable over the period considered. No significant association between the level of ABO antibodies and SARS-CoV-2-specific antibodies was observed (-0.44 < rho < 0.42, p > 0.053). Total IgA in saliva was lower in convalescents than in controls (p = 0.038). Summary/Conclusions: We observed consistently lower serum concentrations of anti-A and anti-B in COVID-19 convalescents than in healthy controls, suggesting ABO antibodies to conferring a degree of protection against SARS-CoV-2 infection. There may be an increased susceptibility to SARS-CoV-2 infection due to individual preexisting low ABO antibody levels. However, the mechanism underlying our observation of significantly reduced ABO antibodies in the serum, but not in the saliva of affected individuals remains unresolved. Further studies to better understand the molecular mechanism underlying our observation are needed.

Reasons for deferral of COVID-19 convalescent plasma donors

Background: Taking into consideration historic success of convalescent plasma in prophylaxis and treatment of various infectious diseases over the century and expansion of COVID-19 pandemics, we started the COVID-19 convalescent plasma program in our country. Aims: The aim of our study was to show the reasons for deferral of the COVID-19 convalescent plasma (CCP) donors. Methods: This is a prospective study organized in the Institute for Transfusion Medicine of Republic of North Macedonia since 30 April 2020 till July 2021. CCP donors eligible for donation were donors without a history of blood transfusion, female donors who have never been pregnant, or who have been tested and found negative for anti- HLA antibodies, age 18-65, in good health that fulfil all other criteria for regular blood donors. All potential donor were tested for: negative RT-PCR for SARSCoV-2 before donation, anti-SARS-CoV-2 antibodies, anti-HLA antibodies (where applicable), blood count, blood group, TTI and biochemistry. Antibody testing was performed at the Institute for Immunobiology and Human Genetics in Skopje using CLIA method with Snibe Maglumi SARS-CoV-2 S-RBD IgG with IgG cut-off for CCP donation larger than 5 AU/ml (cut-off for regular positive result was larger than 1 AU/ml). All donors signed inform consent for donation and inclusion in the study. Results: There were 1462 potential CCP donors, of which 762(52.1%) did not fulfil criteria for CCP donation, 424(55.6%) women and 338 (44.4%) men. There were 454(59.6%) potential CCP donors that did not have enough anti-SARS-CoV-2 S-RBD IgG antibodies (Ab), of which 223(49.1%) had Ab concentration less than 1 AU/ml (29.3% of all deferred donors) and 231(50.9%) had Ab concentration between 1 and 5 AU/ml (30.3% of all deferred donors). In this subgroup of deferred donors, there were 227 women (50%) and 227 men (50%), with mean age 37.2 ± 10.1 (range 18-63). There were statistical significant correlation between the gender and the Ab concentration less than 1 AU/ml and from 1 to 5 AU/ml (Pearson Chi-square: 3.88667, df = 1, p = 0.048671). Multiple regression analysis showed that gender is independently connected with the Ab concentration, OR = 1.4495 (95%CI 1.0016-2.0976), i.e. male gender increases the chance for increased Ab concentration for one and a half times. According to the age group, the majority of deferred CCP donors, because of low Ab concentration, are in the age group from 30 till 39(39.6%), followed by age group from 40 till 49(23.8%), and least deferred donors were in 50 and above age group (13.2%). The most of these deferred donors were treated at home, 451(99.3%) and had symptoms 240(52.9%). Distribution according to blood group in CCP donors deferred due to low Ab concentration was: blood group A- 182(40.1%), blood group O-153(33.7%), blood group B-69(15.1%) and blood group AB-42(9.3%). In the whole investigated group, 65 (8.5%) potential donors were deferred because of low haemoglobin level (less than 12.5 g/dl for women and less than 13.5 g/dl for men), 26 (3.4%) were deferred because of positive TTI markers and 102 (13.4%) women of the total number of deferred donors were deferred because of presence of HLA antibodies (i.e. 24% of investigated women). Summary/Conclusions: There is large percentage of deferred CCP donors mostly because of low Ab concentration, presence of HLA antibodies and low haemoglobin level, but starting of COVID-19 convalescent plasma program was a big success for our institution and our country and helped a lot of patients.

Collection and quality characteristics of Coronavirus Disease 2019 convalescent plasma

Objective To explore the application and safety of apheresis technology in collection of Coronavirus Disease 2019 (COVID-19) convalescent plasma (CP), and to analyze the quality characteristics of the plasma. Methods The general data of COVID-19 convalescent plasma (CP) donors, including gender, age, date of discharge or release from medical isolation, were collected based on informed consent. After physical examination, the CP was collected by apheresis technology with plasma separator, inactivated with methylene blue, and determined for severe acute respiratory symptom Coronavirus 2 (SARS-CoV-2) nucleic acid and specific antibody (RBD-IgG) against SARS-CoV-2. Results The collection process went well, and no serious adverse events related to plasma collection were reported during or after the collection. The average age of COVID-19 CP donors was 38 years (n = 933). The distributions of blood groups A, B, AB and 0 in RhD (+) COVID-19 CP were 33. 4%, 29. 2%, 10% and 27. 2% respectively. The plasma donation date was 18 d from the discharge date in average. All the test results of SARS-CoV-2 nucleic acid in CP were negative, while the proportion of plasma samples at SARS-CoV-2 antibody titer of more than 1: 160 was 92. 60%. Conclusion Apheresis technology was safe and reliable. The COVID-19 CP contained high titer antibody. Large-scale collection and preparation of inactivated plasma against SARS-CoV-2 played an important role in the treatment of COVID-19.