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Microcolumn gel immunoassay (MGIA) has the ability to meet the requirements of clinical diagnosis due to its reliable sensitivity and accuracy. However, traditional MGIA exhibits limitations including inadequate portability, low throughput, and extended analysis time. To address these challenges, we combined MGIA with microfluidic technology, demonstrating a centrifugal microfluidic-based microcolumn gel immunoassay (μMGIA) platform for blood typing of clinical samples. Experimental results indicate that the μMGIA platform can simultaneously detect six blood group antigens in five clinical blood samples within 2 min. Notably, it offers comprehensive detection of ABO blood group antigens and Rh blood group antigens with 100 % accuracy, outperforming the traditional slide method. The integration of microfluidic technology with MGIA circumvents the constraints of traditional methods, providing a new avenue for blood typing and immunoanalysis of clinical samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Human milk oligosaccharides (HMOs) have attracted increasing attention due to the emerging evidence of their positive roles for infant's health. A high-throughput method for absolute quantitation of the complex HMOs including multiple isomeric structures is important but very challenging, due to the highly divers nature and wide variation in content of HMOs from different individuals. Here we used UPLC-MS-MRM in the negative-ion mode for accurate quantitation of 23 complex HMOs in just 15 min. The selected oligosaccharides are in their native forms and include neutral and sialylated, fucosylated and non-fucosylated, linear and branched, and secretor and Lewis phenotype indicators. The well validated method with good sensitivity, recovery and reproducibility was then applied to a large population quantitative survey of 251 Chinese mothers from five different ethnic groups (Han, Zhuang, Hui, Mongolian and Tibetan) living in different geographical regions for their secretor's status and Lewis phenotypes., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The gastrointestinal microbiota members produce α-l-fucosidases that play key roles in mucosal, human milk, and dietary oligosaccharide assimilation. Here, 36 open reading frames (ORFs) coding for putative α-l-fucosidases belonging to glycosyl hydrolase family 29 (GH29) were identified through metagenome analysis of breast-fed infant fecal microbiome. Twenty-two of those ORFs showed a complete coding sequence with deduced amino acid sequences displaying the highest degree of identity with α-l-fucosidases from Bacteroides thetaiotaomicron, Bacteroides caccae, Phocaeicola vulgatus, Phocaeicola dorei, Ruminococcus gnavus, and Streptococcus parasanguinis. Based on sequence homology, 10 α-l-fucosidase genes were selected for substrate specificity characterization. The α-l-fucosidases Fuc18, Fuc19A, Fuc35B, Fuc39, and Fuc1584 showed hydrolytic activity on α1,3/4-linked fucose present in Lewis blood antigens and the human milk oligosaccharide (HMO) 3-fucosyllactose. In addition, Fuc1584 also hydrolyzed fucosyl-α-1,6- N -acetylglucosamine (6FN), a component of the core fucosylation of N -glycans. Fuc35A and Fuc193 showed activity on α1,2/3/4/6 linkages from H type-2, Lewis blood antigens, HMOs and 6FN. Fuc30 displayed activity only on α1,6-linked l-fucose, and Fuc5372 showed a preference for α1,2 linkages. Fuc2358 exhibited a broad substrate specificity releasing l-fucose from all the tested free histo-blood group antigens, HMOs, and 6FN. This latest enzyme also displayed activity in glycoconjugates carrying lacto- N -fucopentaose II (Le a ) and lacto- N -fucopentaose III (Le x ) and in the glycoprotein mucin. Fuc18, Fuc19A, and Fuc39 also removed l-fucose from neoglycoproteins and human α-1 acid glycoprotein. These results give insight into the great diversity of α-l-fucosidases from the infant gut microbiota, thus supporting the hypothesis that fucosylated glycans are crucial for shaping the newborn microbiota composition. IMPORTANCE α-l-Fucosyl residues are frequently present in many relevant glycans, such as human milk oligosaccharides (HMOs), histo-blood group antigens (HBGAs), and epitopes on cell surface glycoconjugate receptors. These fucosylated glycans are involved in a number of mammalian physiological processes, including adhesion of pathogens and immune responses. The modulation of l-fucose content in such processes may provide new insights and knowledge regarding molecular interactions and may help to devise new therapeutic strategies. Microbial α-l-fucosidases are exoglycosidases that remove α-l-fucosyl residues from free oligosaccharides and glycoconjugates and can be also used in transglycosylation reactions to synthesize oligosaccharides. In this work, α-l-fucosidases from the GH29 family were identified and characterized from the metagenome of fecal samples of breastfed infants. These enzymes showed different substrate specificities toward HMOs, HBGAs, naturally occurring glycoproteins, and neoglycoproteins. These novel glycosidase enzymes from the breast-fed infant gut microbiota, which resulted in a good source of α-l-fucosidases, have great biotechnological potential.

Background: COVID 19 infection caused by novel coronavirus with no specific established treatment. Convalescent Plasma Therapy has been authorized as an off-label therapeutic procedure. We assessed the outcome of convalescent plasma (CP) units versus standard treatment on the complete recovery, improvement and 28 days' mortality of COVID 19 patients., Materials and Methods: The present was multi-centric case controlled observational prospective study. The study was conducted for a period of four and half months from July 15 2020 to 30 November 2020 after taking approval from the Expert Committee, Health & Family Welfare Department, Government of Odisha. Plasma therapy was applied on two groups of 1189 serious COVID patients (959 number of pre- critical and 230 number of critical patients) not responding to oxygen therapy. It was compared with non- transfused control group of 1243 patients (996 number of pre-critical and 247 number of critical patients)., Results: Discharge was better in (55.5%) transfused than (43%)in non-transfused pre-critical patients and the mortality was lower (44.3%) in transfused, (48.9%) than non-transfused critical patients respectively. Complete recovery was highest in those who were transfused with CP with neutralizing titer more than 1:160 (52.5%), 18-30 years' age group (64%), females (53%), 'O' Rh D positive blood group (51.5%). There was no adverse reaction due to CP transfusion., Conclusions: CP is effective in improving the recovery rate with earlier discharge and decrease in the 28 days' mortality than in the control non-transfused group. CP with neutralizing antibody titer more than 1:160 has the best outcome with complete recovery and decrease in the mortality. It is more effective in treating pre-critical patients when transfused early, in female patients, in younger age group and in blood group 'O' Rh D positive., (Copyright © 2021 Société française de transfusion sanguine (SFTS). Published by Elsevier Masson SAS. All rights reserved.)

Objectives: To verify the frequency of viruses causing acute gastroenteritis (AGE) in association with the histo-blood group antigen (HBGA) and Rotarix™ vaccination coverage in children from the Amazon region., Design: Fecal and saliva samples were collected from children with AGE (n = 485) and acute respiratory infection (ARI) (n = 249) clinical symptoms. Rotavirus A (RVA), norovirus, human adenovirus (HAdV), and sapovirus (SaV) were verified in feces by molecular detection. Saliva samples were used for HBGA phenotyping/FUT3 genotyping. Blood group types, clinical aspects and Rotarix™ RVA vaccination data were recorded., Results: Norovirus remained the most prevalently detected cause of AGE (38%, 184/485 and ARI 21.3%, 53/249). High HAdV frequencies were observed in AGE children (28.6%, 139/485) and ARI children (37.3%, 93/249). RVA was the third most prevalent virus causing AGE (22.7%, 110/485 and ARI 19.3%, 48/249) and a low RV1 coverage (61%, 448/734) was verified. The SaV frequencies were lower (7.2%, 35/485 for AGE and 6.8%, 17/249 for ARI). Secretor children were HBGA susceptible to HAdV infection (OR 1.5, 95% CI 1.0-2.3; P = 0.04) but not to RVA, norovirus or SaV infection., Conclusions: Norovirus could be considered the main etiological agent of AGE. No association was verified for HBGA susceptibility to RVA, norovirus and SaV. Secretor children showed a slight susceptibility to HAdV infection and the Le (a-b-) heterogeneous SNPs on the FUT3 gene., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In a context where SARS-CoV-2 population-wide testing is implemented, clinical features and antibody response in those infected have never been documented in Africa. Yet, the information provided by analyzing data from population-wide testing is critical to understand the infection dynamics and devise control strategies. We described clinical features and assessed antibody response in people screened for SARS-CoV-2 infection. We analyzed data from a cohort of 3464 people that we molecularly screened for SARS-CoV-2 infection in our routine activity. We recorded people SARS-CoV-2 diagnosis, age, gender, blood types, white blood cells (WBC), symptoms, chronic disease status and time to SARS-CoV-2 RT-PCR conversion from positive to negative. We calculated the age-based distribution of SARS-CoV-2 infection, analyzed the proportion and the spectrum of COVID-19 severity. Furthermore, in a nested sub-study, we screened 83 COVID-19 patients and 319 contact-cases for anti-SARS-CoV-2 antibodies. Males and females accounted for respectively 51% and 49% of people screened. The studied population median and mean age were both 39 years. 592 out of 3464 people (17.2%) were diagnosed with SARS-CoV-2 infection with males and females representing, respectively, 53% and 47%. The median and mean ages of SARS-CoV-2 infected subjects were 37 and 38 years respectively. The lowest rate of infection (8%) was observed in the elderly (aged > 60). The rate of SARS-Cov-2 infection in both young (18-35 years old) and middle-aged adults (36-60 years old) was around 20%. The analysis of SARS-CoV-2 infection age distribution showed that middle-aged adults accounted for 54.7% of SARS-CoV-2 positive persons, followed respectively by young adults (33.7%), children (7.7%) and elderly (3.8%). 68% (N = 402) of SARS-CoV-2 infected persons were asymptomatic, 26.3% (N = 156) had influenza-like symptoms, 2.7% (N = 16) had influenza-like symptoms associated with anosmia and ageusia, 2% (N = 11) had dyspnea and 1% (N = 7) had respiratory failure, which resulted in death. Data also showed that 12% of SARS-CoV-2 infected subjects, had chronic diseases. Hypertension, diabetes, and asthma were the top concurrent chronic diseases representing respectively 58%, 25% and 12% of recorded chronic diseases. Half of SARS-CoV-2 RT-PCR positive patients were cured within 14 days following the initiation of the anti-COVID-19 treatment protocol. 78.3% of COVID-19 patients and 55% of SARS-CoV-2 RT-PCR confirmed negative contact-cases were positive for anti-SARS-CoV-2 antibodies. Patients with severe-to-critical illness have higher leukocytes, higher neutrophils and lower lymphocyte counts contrarily to asymptomatic patients and patients with mild-to-moderate illness. Neutrophilic leukopenia was more prevalent in asymptomatic patients and patients with mild-to-moderate disease for 4 weeks after diagnosis (27.1-42.1%). In Patients with severe-to-critical illness, neutrophilic leukocytosis or neutrophilia (35.6-50%) and lymphocytopenia (20-40%) were more frequent. More than 60% of participants were blood type O. It is also important to note that infection rate was slightly higher among A and B blood types compared with type O. In this African setting, young and middle-aged adults are most likely driving community transmission of COVID-19. The rate of critical disease is relatively low. The high rate of anti-SARS-CoV-2 antibodies observed in SARS-CoV-2 RT-PCR negative contact cases suggests that subclinical infection may have been overlooked in our setting.

Background and Objectives: Next generation sequencing (NGS) has promising applications in transfusion medicine. Exome sequencing (ES) is increasingly used in the clinical setting, and blood group interpretation is an additional value that could be extracted from existing data sets. We provide the first release of an open-source software tailored for this purpose and describe its validation with three blood group systems., Materials and Methods: The DTM-Tools algorithm was designed and used to analyse 1018 ES NGS files from the ClinSeq ® cohort. Predictions were correlated with serology for 5 antigens in a subset of 108 blood samples. Discrepancies were investigated with alternative phenotyping and genotyping methods, including a long-read NGS platform., Results: Of 116 genomic variants queried, those corresponding to 18 known KEL, FY and JK alleles were identified in this cohort. 596 additional exonic variants were identified KEL, ACKR1 and SLC14A1, including 58 predicted frameshifts. Software predictions were validated by serology in 108 participants; one case in the FY blood group and three cases in the JK blood group were discrepant. Investigation revealed that these discrepancies resulted from (1) clerical error, (2) serologic failure to detect weak antigenic expression and (3) a frameshift variant absent in blood group databases., Conclusion: DTM-Tools can be employed for rapid Kell, Duffy and Kidd blood group antigen prediction from existing ES data sets; for discrepancies detected in the validation data set, software predictions proved accurate. DTM-Tools is open-source and in continuous development., (© 2020 International Society of Blood Transfusion.)

Stress has a major impact on the modulation of metabolism, as previously evidenced by hyperglycemia following chronic social defeat (CSD) stress in mice. Although CSD-triggered metabolic dysregulation might predispose to pre-diabetic conditions, insulin sensitivity remained intact, and obesity did not develop, when animals were fed with a standard diet (SD). Here, we investigated whether a nutritional challenge, a high-fat diet (HFD), aggravates the metabolic phenotype and whether there are particularly sensitive time windows for the negative consequences of HFD exposure. Chronically stressed male mice and controls (CTRL) were kept under (i) SD-conditions, (ii) with HFD commencing post-CSD, or (iii) provided with HFD lasting throughout and after CSD. Under SD conditions, stress increased glucose levels early post-CSD. Both HFD regimens increased glucose levels in non-stressed mice but not in stressed mice. Nonetheless, when HFD was provided after CSD, stressed mice did not differ from controls in long-term body weight gain, fat tissue mass and plasma insulin, and leptin levels. In contrast, when HFD was continuously available, stressed mice displayed reduced body weight gain, lowered plasma levels of insulin and leptin, and reduced white adipose tissue weights as compared to their HFD-treated non-stressed controls. Interestingly, stress-induced adrenal hyperplasia and hypercortisolemia were observed in mice treated with SD and with HFD after CSD but not in stressed mice exposed to a continuous HFD treatment. The present work demonstrates that CSD can reduce HFD-induced metabolic dysregulation. Hence, HFD during stress may act beneficially, as comfort food, by decreasing stress-induced metabolic demands.

Objective: There are few published researches on blood groups, hematological parameters [hemoglobin, red cell distribution width (RDW), white blood cells (WBCs), mean platelets volume (MPV)] and gestational diabetes mellitus (GDM). The aim of this study was to investigate the association of haematological indices with GDM in early pregnancy., Methods: The study was carried out at Saad Abuelela Hospital (Khartoum, Sudan) during March-November of 2018. Pregnant Sudanese women in early pregnancy (gestational age <14 weeks) were enrolled in the study. The details of the medical and obstetrics history were recorded. The women were then followed up until 24-28 weeks of gestation when a 75-gram oral glucose tolerance test was performed., Results: Two hundred and fifty-three women at 10.2 week of gestational age completed the follow-up. The mean (SD) of the age and gravidity at the initial antenatal visit were 28.03 (5.6) years, 2.32 (2.41). The mean (SD) of body mass index (BMI) was 27.28 (24.41-30.80) kg/m 2 . Fifty women (19.8%) had GDM. Age, parity, BMI, place of residence, employment and education were not significantly different between the two groups. Moreover, there was no significant difference in the blood groups and hematological parameters between women with and without GDM., Conclusion: In this study, the blood groups and other hematological parameters were not different between women with and without GDM., (Copyright © 2021 by the Association of Clinical Scientists, Inc.)

Background: This report evaluates hospital blood use trends during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, and identifies factors associated with the need for transfusion and risk of death in patients with coronavirus 2019 (COVID-19)., Methods: Overall hospital blood use and medical records of adult patients with COVID-19 were extracted for two institutions. Multivariate logistic regression models were conducted to estimate associations between the outcomes transfusion and mortality and patient factors., Results: Daily blood use decreased compared to pre-COVID-19 levels; the effect was more significant for platelets (29% and 34%) compared to red blood cells (25% and 20%) at the two institutions, respectively. Surgical and oncologic services had a decrease in average daily use of platelets of 52% and 30%, and red blood cells of 39% and 25%, respectively. A total of 128 patients with COVID-19 were hospitalized, and 13 (10%) received at least one transfusion due to anemia secondary to chronic illness (n = 7), recent surgery (n = 3), and extracorporeal membrane oxygenation (n = 3). Lower baseline platelet count and admission to the intensive care unit were associated with increased risk of transfusion. The blood group distribution in patients with COVID-19 was 37% group O, 40% group A, 18% group B, and 5% group AB. Non-type O was not associated with increased risk of mortality., Conclusion: The response to the SARS-CoV-2 pandemic included changes in routine hospital operations that allowed for the provision of a sufficient level of care for patients with and without COVID-19. Although blood type may play a role in COVID-19 susceptibility, it did not seem to be associated with patient mortality., (© 2020 AABB.)

Background: The ABO and the Rhesus blood group systems, as well as various abnormal haemoglobin (Hb) variants (haemoglobinopathies) are known to influence malaria parasite carriage and disease severity in individuals living in malaria endemic areas. This study identified the blood group and Hb variant distribution and Plasmodium falciparum infection status of afebrile individuals living in southern Ghana., Methods: Afebrile participants were recruited from Obom (358) in the Greater Accra Region and Ewim (100) and Simiw (329) in the Central Region of Ghana. Venous blood (1 ml) was collected into EDTA vacutainer tubes. Three 20 μl drops of blood were used for blood group analysis using the tile method. Another 500 μl aliquot was used for the qualitative sickling test using sodium metabisulphite and haemoglobin electrophoresis. Genomic DNA was extracted from 100 μl of whole blood and used in P. falciparum species-specific PCR., Results: The most abundant blood group and abnormal haemoglobin variant in both sites was blood group O + (47.4%) and HbAS (15.8%). A total of 13 (1.7%) of the participants had full haemoglobinopathies (SS, SC and CC), whilst 196 (25.4%) were carriers (AS and AC). Although there was a significantly higher prevalence of sickling positive participants from the Central Region, genotyping identified a similar prevalence of each of the abnormal haemoglobin genes in both sites. Asymptomatic parasite carriage estimated by PCR was 40.9% in the Central Region and 41.8% in the Greater Accra Region., Conclusions: Asymptomatic carriage of P. falciparum parasite in the study population was not associated with any particular blood group variant or haemoglobin genotype.

Rotaviruses cause severe gastroenteritis in infants, in which the viruses interact with human histo-blood group antigens (HBGAs) as attachment and host susceptibility factors. While gastroenteritis outbreaks caused by rotaviruses are uncommon in adolescents, we reported here one that occurred in a middle school in China. Rectal swabs and saliva samples were collected from symptomatic and asymptomatic students, and samples were also collected from the environment. Using PCR, followed by DNA sequencing, a single G9P[8] rotavirus strain was identified as the causative agent. The attack rate of the outbreak was 13.5% for boarders, which was significantly higher than that of day students (1.8%). Person-to-person transmission was the most plausible transmission mode. The HBGA phenotypes of the individuals in the study were determined by enzyme immunoassay, using saliva samples, while recombinant VP8* protein of the causative rotavirus strain was produced for HBGA binding assays to evaluate the host susceptibility. Our data showed that secretor individuals had a significantly higher risk of infection than nonsecretors. Accordingly, the VP8* protein bound nearly all secretor saliva samples, but not those of nonsecretors, explaining the observed infection of secretor individuals only. This is the first single-outbreak-based investigation showing that P[8] rotavirus infected only secretors. Our investigation also suggests that health education of school students is an important countermeasure against an outbreak of communicable disease.

We report the fabrication of silver nanoparticles (AgNPs) surface functionalized with gelatin at different concentrations (G10/G20/G40 AgNPs) with an average particle size of ∼200 nm, bioconjugated with antisera antibodies (AsAbs) of the major and clinically significant blood groups (CSBGs) at different titres from neat to 1:128. Bioconjugation using ionic interaction at pH 7.4 enabled 'end-on' configuration, with the -NH2 group of the antibody free for interaction with the red blood cell antigen, as confirmed by Fourier transform infrared spectroscopy. The tube agglutination test (TAT) revealed optimum agglutination with G20NPs, while SDS PAGE confirmed the optimal titre as 1:8 for the major blood groups A, B, AB and O. Bioconjugated AgNPs coated onto microtitre assay plates with the major blood groups and CSBGs to enable simultaneous identification, were validated against the TAT on 400 random blood samples for the major blood groups and revealed high accuracy (95%). While similar accuracy was seen for most of the CSBGs with only false negatives, the method was not found to be suitable for the Kell, Kidd and Duffy groups. The absence of false positives reflects high safety, and eliminates the risk of a mismatched blood transfusion. The method uses diluted blood and hence could enable point-of-care detection. The significantly lower AsAb requirement also provides a cost advantage.

Background: Several approaches are used by blood centers when providing minor (non-ABO/D) antigen-negative RBCs to hospitals. Details vary but include providing results on the unit labeling intended for clinical use without retyping or providing results on packing documents or via computer query requiring confirmation. Recent regulatory changes allow labeling with historical minor antigen results, defined as previously performed by the donor center on two different donations with results linked to the donor and confirmed concordant. Here we investigate causes of discrepancies and identify critical process steps., Study Design and Methods: Nine years (2009-2017) of data were reviewed for number, antigen system, and root cause of discrepancies flagged by the computer when retyping donors prior to labeling (internal discrepancies) or reported by the hospital when retested (external discrepancies). Licensed automated (CcEeK) and tube methods were used., Results: Among 300,000 samples phenotyped for CcEe, K, Fy a/b , Jk a/b , Ss (>3 million antigens), ∼1,389,960 were repeated on 2nd donation with 397 (1/3501) discordant; 205 Fy, 118 Rh, and 74 others. Of ∼682,691 antigen-negative phenotypes provided on unit labeling, ∼37.5% (256,118) were retyped by hospitals with 29 discrepancies (1/8832), primarily Rh variants., Conclusion: When repeating minor antigen types by serology, discrepancies are primarily associated with weak Fy b , among Caucasian donors, and weak/partial Rh antigens in donors of African ancestry. DNA-based testing avoids these. To label with historical results, accuracy is increased by automated testing with direct computer interface. Testing on two donations with results confirmed to be concordant is not inferior to testing on the current donation., (© 2020 AABB.)

Blood group types are associated with coronary artery disease. However, data are scarce about the impact of blood group types on coronary collateral circulation. In this study, we aimed to investigate the relationship between the blood group types and coronary collateral circulation. Two hundred and twelve patients who underwent coronary angiography in our department and had a stenosis of ≥ 90% in at least one major epicardial vessel were included in our study. Collateral degree was graded according to Rentrop-Cohen classification. After grading, patients were divided into poor coronary collateral circulation (Rentrop grade 0 and 1) and good coronary collateral circulation (Rentrop 2 and 3) groups. The ABO blood type of all participants was determined. The incidence rates of O blood group type were significantly higher in the good coronary collateral group compared to the poor collateral group (37.9% vs 17.1%, P < .001). The O type blood group was an independent predictor of good coronary collateral circulation (odds ratio = 1.83, 95% confidence interval = 1.56-6.18, P = .015). Coronary collateral circulation is associated with blood group types. The O blood group predicts good coronary collateral development among patients with coronary artery disease.

Background: The appropriate cutoff to define a positive point-of-care card agglutination (CA) test for dog erythrocyte antigen 1 (DEA 1) blood typing depends on whether the test can be used in the donor or recipient., Objectives: By screening for CA test positivity, we aimed to evaluate the best cutoff value for DEA 1 blood typing in canine blood donors using a receiver operator characteristic (ROC) curve., Methods: Ethylenediaminetetraacetic acid (EDTA) blood samples from 100 canine blood donors were blood-typed in parallel for DEA 1 using both immunochromatographic (IC) and CA tests. The effect of temperature, storage time, and anticoagulant solutions for both methods was evaluated. Unweighted and weighted Cohen's Kappa (K) statistic was calculated to evaluate the agreement between the two testing methods. The overall performance of the CA test was evaluated by generating a ROC curve using the IC test as the reference method., Results: Concordant results were obtained for 86% of the samples. Unweighted and weighted K statistics demonstrated good and moderate agreement, respectively. For the CA test, the ROC curve showed an area under the curve (AUC) of 0.910, with the highest sensitivity cutoff values at ≥1+ agglutination. CA- and IC-typed EDTA blood samples stored at room temperature for up to 1 week and refrigerated for up to 1 month were concordant as were the citrate phosphate dextrose adenine 1 (CPDA-1) anticoagulated blood samples stored for up to 1 week at 4 ± 2°C., Conclusions: The overall reliability of the CA method seemed to be lower than that of the IC method. When CA is used as a screening test for canine blood donors, the correct cut off is ≥1+ agglutination is recommended to maximize sensitivity., (© 2019 American Society for Veterinary Clinical Pathology.)

Background: intestinal glycocalyx plays a role in bacterial translocation as the pathogenesis sepsis derived from intra-abdominal infections that vulnerable in certain blood types. However, the link between intestinal glycocalyx in specific types of blood groups and abdominal infections remains unknown. This study aims to find out the condition of intestinal glycocalyx in certain blood types with intraabdominal sepsis., Methods: descriptive study involved subjects with intraabdominal infections who underwent laparotomy. Samples are in the form of intestinal specimens. The measurement of intestinal glycocalyx proceeded by the ELISA method using blood group antigens (A and B). Expression data on the secretors were analyzed using the Kolmogorov - Smirnov test followed by parametric comparisons using ANOVA and t-tests., Results: there were 32 subjects with intra-abdominal infections studied in this study. All of them are secretors and express A and B antigens strongly. We found no difference between intraabdominal infections in those with complications or without complications. Blood type O is a predominant blood type found (43.8%). Escherichia coli is the most commonly found microbe in the culture (61.3%)., Conclusion: this study shows there is no disrupted intestinal glycocalyx of sepsis patients caused by intraabdominal infection.

Donor red cell genotyping provides efficiencies to identify "in demand" blood donors for transfusion recipients requiring antigen-negative blood. Donor red cell genotype information can be used to label units with historical types and introduced throughout the supply chain from the blood center to the hospital transfusion service and has potential to be used in recruitment strategies., (© 2019 AABB.)

Introduction : Classic methods to determine human red blood cell (RBC) antigens are based on serologic testing, however there are some limitations. Based on these, several molecular techniques have been developed to bypass these restrictions and to detect the most important allelic variants, the last of great value for the approach in the personalized medicine, especially for patients in chronic transfusion regime. Areas covered : In this article, we provide an updated review of the ID CORE XT (Progenika Biopharma-Grifols, Bizkaia, Spain) that is one of the commercial DNA-based techniques available to genotyping of RBC antigens. Expert opinion : It is a platform based on Luminex technology with an automated workflow after DNA extraction and its robustness has been shown through validation studies. This technology provides a useful tool to increase the inventory of antigen-negative RBC units, through a mass-scale donor genotyping for RBC antigens and prevent immunization of patients who require chronic transfusion by providing compatible RBC units based on matching by DNA testing.

Background: After-hours or out-of-clinic crossmatches are often limited by the lack of access to specialized material and technical expertise., Hypothesis/objectives: The goal was to adapt a stall-side crossmatch test for pretransfusion evaluation in horses., Animals: Twelve healthy mares (plasma and blood donors, teaching mares)., Methods: In a prospective study, blood from 12 mares was used to compare the results of 132 crossmatches performed with a rapid gel assay to crossmatches performed with a microgel column assay, and with predicted compatibilities based on blood types and detection of antibodies at a reference laboratory (microplate assay). The rapid gel assay protocol for dogs was adapted to decrease the formation of rouleaux that initially precluded equine erythrocytes migration through the gel., Results: There was a good agreement between the rapid gel assay and the microgel assay as well as with the predicted compatibilities (κ > .6 for both). Agreement was higher between the microgel assay and the predicted compatibilities (κ = .8). The rapid gel assay failed to detect 6 predicted Aa incompatibilities (agglutinins-related), 3 of which were also not detected with the microgel assay., Conclusions and Clinical Importance: Based on these results, the modified rapid gel assay could be useful in settings when access to the microgel assay is not available. Discrepancies between both gel techniques and predicted compatibilities were most often low-grade agglutination, which warrants further investigation to assess their clinical importance., (© 2019 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

Conclusions: This update of the Scianna blood group system (Brunker PA, Flegel WA. Scianna: the lucky 13th blood group system. Immunohematology 2011;27:41-57) provides the recent work on the genetic variation of ERMAP across more world populations, the elucidation of the molecular basis of an historical serologic case, new cases of antibodies in the system, the development of new serologic reagents, and new discoveries in the biology of the erythroid membrane associated protein (ERMAP). Although genetic variation in ERMAP has been extensively cataloged, nonsynonymous variants associated with alloantigens have remained limited, and no new antigens have been identified. The first case of a severe hemolytic transfusion reaction to anti-Sc2 has recently been reported, highlighting the importance of pursuing the possibility of antibodies to low-prevalence antigens via indirect antiglobulin testing as a routine component of all transfusion reaction investigations. The expanding use of molecular testing in blood centers and transfusion services has uncovered a wider population distribution of Scianna antigens and heightened the awareness of this blood group system. The International Society of Blood Transfusion recognizes seven antigens in the Scianna blood group system 13.

Background: Graft immunocomplex capture fluorescence analysis is an attractive method to detect intragraft donor-specific anti-HLA antibodies. In ABO-incompatible transplantation, anti-A and B antibodies are also considered to be important donor specific antibodies (ABO-DSA). Therefore, it is useful to monitor intragraft ABO-DSAs to assess antibody-mediated rejection., Methods: To capture A and B antigens, anti-Band III, von Willebrand factor (VW), and plasmalemma vesicle-associated protein (PLVAP) beads were produced. The allograft specimen was homogenized in a lysis buffer. Subsequently, A and B antigens were captured by anti-Band III, VW, or PLVAP beads. The immune complexes were then detected by phycoerythrin-conjugated anti-human IgG antibodies and analyzed using a Luminex system., Results: Although Band III and VW beads yielded false positives and false negatives, PLVAP beads captured A and B antigens with high sensitivity (91.7%) and specificity (100%) when an index > 1.5 was considered positive. The proximity in A and B antigens and PLVAP expression was confirmed using immunohistochemical evaluation. Furthermore, sodium dodecyl sulfate polyacrylamide gel electrophoresis supported that PLVAP is an A and B antigen carrier protein., Case Report: Biopsies were conducted following an ABO-incompatible renal transplant (type A to O) and evaluated for ABO-DSA. Graft immunocomplex capture fluorescence analysis was demonstrated as follows: 3.19 (1 h, serum creatinine [s-Cr] 3.95 mg/dL, titer IgG 1:512, glomerulitis [g] 0, peritubular capillaritis [ptc] 0, complement 4d [C4d] 1); 1.8 (4 d, s-Cr 2.29 mg/dL, titer 1:256, g 0, ptc 0, C4d 3); 1.2 (22 d, s-Cr 1.58 mg/dL, titer 1:128, g 0, ptc 2, C4d 3). This result indicated that the remnant ABO-DSA were adsorbed and subsequently removed from the allograft successfully., Conclusions: This novel application could be used to detect intragraft ABO-DSAs, which could lead to a correct diagnosis and shed light on the ABO-DSA kinetics following ABO-incompatible transplantation., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Genomics is affecting all areas of medicine. In transfusion medicine, DNA-based genotyping is being used as an alternative to serological antibody-based methods to determine blood groups for matching donor to recipient. Most antigenic polymorphisms are due to single nucleotide polymorphism changes in the respective genes, and DNA arrays that target these changes have been validated by comparison with antibody-based typing. Importantly, the ability to test for antigens for which there are no serologic reagents is a major medical advance to identify antibodies and find compatible donor units, and can be life-saving. This review summarizes the evolving use and applications of genotyping for red cell and platelet blood group antigens affecting several areas of medicine. These include prenatal medicine for evaluating risk of fetal or neonatal disease and candidates for Rh-immune globulin; transplantation for bone marrow donor selection and transfusion support for highly alloimmunized patients and for confirmation of A2 status of kidney donors; hematology for comprehensive typing for patients with anemia requiring chronic transfusion; and oncology for patients receiving monoclonal antibody therapies that interfere with pretransfusion testing. A genomics approach allows, for the first time, the ability to routinely select donor units antigen matched to recipients for more than ABO/RhD to reduce complications. Of relevance, the growth of whole-genome sequencing in chronic disease and for general health will provide patients' comprehensive extended blood group profile as part of their medical record to be used to inform selection of the optimal transfusion therapy., (© 2019 by The American Society of Hematology.)

Group A rotaviruses are a major cause of acute gastroenteritis in children. The diversity and unequal geographical prevalence of rotavirus genotypes have been linked to histo-blood group antigens (HBGAs) in different human populations. In order to evaluate the role of HBGAs in rotavirus infections in our population, secretor status (FUT2+), ABO blood group, and Lewis antigens were determined in children attended for rotavirus gastroenteritis in Valencia, Spain. During three consecutive years (2013-2015), stool and saliva samples were collected from 133 children with rotavirus infection. Infecting viral genotypes and HBGAs were determined in patients and compared to a control group and data from blood donors. Rotavirus G9P[8] was the most prevalent strain (49.6%), followed by G1P[8] (20.3%) and G12P[8] (14.3%). Rotavirus infected predominantly secretor (99%) and Lewis b positive (91.7%) children. Children with blood group A and AB were significantly more prone to rotavirus gastroenteritis than those with blood group O. Our results confirm that a HBGA genetic background is linked to rotavirus P[8] susceptibility. Rotavirus P[8] symptomatic infection is manifestly more frequent in secretor-positive (FUT2+) than in non-secretor individuals, although no differences between rotavirus G genotypes were found.

Red blood cell transfusions have become an integral component in the treatment of anemic patients and the number of transfusions continues to increase from year to year in both veterinary and human medicine. Although crucial, red blood cell transfusions are not benign and can lead to serious adverse reactions. Therefore, determining the most appropriate donor unit to use for transfusion can be challenging. As transfusion medicine evolves, greater numbers of blood surface antigens are being documented and pretransfusion testing protocols are becoming more complex. In order to better understand this evolving field, this review presents the basic immunology of pretransfusion testing, including methodologies of blood typing and crossmatch testing, and their application in canine and feline medicine., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: Blood-group typing of donors and patients is essential to avoid incompatible transfusions. Transfusion of incompatible RBCs may result in alloimmunization complicating future transfusions or in the presence of antibodies in adverse reactions. With more than 300 blood group antigens identified, it is difficult to provide fully compatible blood. Currently, standard practice is to match for the most immunogenic antigens. While the current agglutination-based RBC-typing methods are reliable for testing a selected number of antigens, they are not easily adaptable for high-throughput multiplex blood typing beyond the current standard., Study Design and Methods: Surface plasmon resonance (SPR) is a label-free method to follow molecular-and, very recently, also cellular-interactions in real time. Demonstration of binding of RBCs to blood group antigen-specific antibodies by SPR has already been achieved. Here, we demonstrate the generation of an SPR array equipped with clinically relevant blood group antibodies (A, B, and Rh blood groups). To validate this method, we blindly compared typing of 946 blood donors with results of current diagnostic agglutination-based methods., Results: RBC typing was achieved by monitoring RBC binding to blood group-specific antibodies on the sensor simultaneously within 5 minutes per sample. Regeneration of the chip was robust, allowing for typing of at least 100 samples. The typing results gave a 100% match with classical serology with all antibodies tested besides anti-E/e monoclonals, which gave inconsistent results due to low antibody specificity., Conclusion: This study demonstrates that SPR-based RBC typing for multiple antigens can be realized simultaneously with high-quality antibodies, enabling reduced hands-on time and possibly improving cost efficiency., (© 2018 AABB.)

Conclusions: This update of the Knops (KN) blood group system (Moulds JM. The Knops blood group system. Immunohematology 2010;26:2-7) adds no new antigens to this system (International Society of Blood Transfusion system 22), which currently has nine antigens. However, the molecular basis of York, KN5, or Yka has been identified as c.4223C>T and designated KN*01.-05. Although not considered clinically significant in the field of transfusion medicine, there has been great interest in the Knops polymorphism by investigators working on malaria documented by numerous studies over the past 8 years.

The maintenance of normal blood flow through the vessels is the result of the coordinated work of the coagulation and anticoagulation systems of our body. The balance of this system depends on many factors, including endothelial, humoral, platelet ones, however, we still lack knowledge about the effect of antigenic determinants on the state of the hemostatic system. This study is devoted to assessing the effect of the presence and absence of antigens on the AB0 system, presented on erythrocyte and platelet membranes, on hemostatic parameters. The study was conducted in the Clinics of Samara State Medical University and consisted of127 clinically healthy individuals who underwent a general analysis and biochemical blood analysis, 52 people with the most stable indicators of cell composition and metabolic profile were selected for a coagulation test, including determination of the activity of coagulation factors and routine tests. A significant decrease in the activity of the VIII and VII coagulation factors was revealed, as well as an increase in the prothrombin time in patients with 0 (I) blood group compared to the "antigenic" blood groups. The presence of biological variation for indicators of external and internal coagulation paths was noted, depending on the group of blood belonging to the AB0 system. The findings suggest that there is an increased susceptibility to bleeding in patients with 0 (I) blood groups due to the absence of antigenic determinants on the cell membrane, while for "antigenic" blood groups, on the contrary, there is a susceptibility to thrombosis due to increased activity of the components of the coagulation system., Competing Interests: The authors declare no conflict of interest.

Conclusions: The presence of multiple alloantibodies or an antibody to a highprevalance antigen in a patient sample can pose challenges in antibody identification. The pattern of reactivity seen on an antibody panel may show various strengths of reactivity by different methods of testing or same strength of reactivity at one or more phases of testing. To ensure proper identification, multiple investigative tools may be used. We review one of these methods-inhibition by soluble substances-which has become an expansion of our toolbox within the past 10 years. Alloantibodies can be inhibited using specific soluble substances. These soluble substances occur naturally in various fluids or can be manufactured. When a patient sample contains multiple antibodies, clinically significant or not, inhibition of one may help determine specificities of others. Specific inhibition of a particular antibody will also help to confirm its presence.

Conclusions: This update of the CD59 blood group system (Weinstock C, Anliker M, von Zabern I. CD59: a long-known complement inhibitor has advanced to a blood group system. Immunohematology 2015;31:145-51) increases the number of reported patients with CD59 deficiency from 10 to 14. All of these 14 patients suffered from severe illness. Recently, a new variant allele was found in heterozygosity. Flow cytometry data suggest that this variant was expressed on the red blood cells of the propositus. Although additional alleles have been found, the CD59 system (International Society of Blood Transfusion system 35) continues to have one antigen.

Conclusions: This update of the Augustine (AUG) blood group system (Daniels G. The Augustine blood group system, 48 years in the making. Immunohematology 2016;32:100-3) describes two antigens that have been added to the Augustine system (International Society of Blood Transfusion system 36), bringing the number of antigens in the system to four. Further information on the clinical significance of Augustine system antibodies and the function of the Augustine glycoprotein, equilibrative nucleoside transporter 1, is presented.

Correct blood group typing is a prerequisite for transfusion. In most cases blood group determination is without problems; however, in individual cases various factors can complicate blood group determination and sometimes lead to confusing findings. For a better understanding the clinician should have basic knowledge of blood typing. Blood group determination usually covers the AB0 blood groups, Rhesus and Kell systems; in addition, a direct Coombs test and an antibody screening test for the detection of irregular antibodies in the recipient are performed. Confusion of patients, blood samples, results or preparations can lead to severe consequences due to incompatible transfusion and must be prevented. In this context, bedside blood type testing before transfusion is of utmost importance. Problems in laboratory analysis as well as patient-related factors, such as the existence of irregular antibodies against red blood cells can complicate the immunohematology diagnostics. Certain medications, such as daratumumab, lead to a significantly increased complexity in laboratory analyses. Massive transfusions can lead to chimerism with more than one population of circulating red blood cells. Hematopoetic stem cell transplantation can also lead to a change in blood groups as well as chimerism. In addition, there are various other rare causes that can result in difficulties in blood group determination, such as rare blood groups or rare disease-associated phenomena. In the case of problems in blood group determination, early and close cooperation with transfusion medicine is essential for the clinician.

Conclusions: This article reviews information regarding the clinical significance of antibodies to antigens in the Scianna, Dombrock, Colton, Landsteiner-Wiener, Chido/Rodgers, H, Kx, Cromer, Gerbich, Knops, Indian, and Ok blood group systems. Like most blood group systems, antibodies to many of the antigens in these groups are rarely encountered because of the high prevalence of the associated antigens in most populations. For many, the clinical significance-that is, the potential to cause reduced survival of transfused antigen-positive red blood cells or a transfusion reaction (e.g., anti-Ge2, anti-H) and/or hemolytic disease of the fetus and newborn (e.g., anti-Coa, anti-Ge3)-has been documented. Some of these antibodies are not always clinically significant, and because of the high prevalence of the antigen, antigen-negative blood may be extremely difficult to find (e.g., anti-LW, anti-Inb). The use of a monocyte monolayer assay may be helpful when making transfusion decisions for patients with these antibodies. For others, their prevalence is so rare that information on the clinical significance of their antibodies is not available (e.g., anti-Co4, anti-Ok).

Conclusions: This article reviews information on the clinical significance of antibodies to antigens in the Raph, John Milton Hagen, I, Globoside, Gill, Rh-associated glycoprotein, FORS, JR, LAN, Vel, CD59, and Augustine blood group systems. Antibodies to many of the antigens in these groups are rarely encountered because of the high prevalence of the associated antigens in most populations. For many of these antibodies, the clinical significance-that is, the potential to cause reduced survival of transfused antigen-positive red blood cells or a transfusion reaction (e.g., anti-P, anti-Jra, and anti-Lan), and/or hemolytic disease of the fetus and newborn (e.g., anti-RHAG4 and anti-Vel)-has been documented. For other antibodies, their prevalence is so rare that information on the clinical significance of their antibodies is not available (e.g., anti-FORS1).

Background: Human noroviruses (HuNoVs) are a prominent cause of gastroenteritis, yet fundamental questions remain regarding epidemiology, diversity, and immunity in sub-Saharan African children. We investigated HuNoV seroprevalence and genetic and sociodemographic risk factors in Ugandan children., Methods: We randomly screened 797 participants of a longitudinal birth cohort (Entebbe, EMaBS) and 378 from a cross-sectional survey (rural Lake Victoria, LaVIISWA), for antibodies against HuNoV genotypes by ELISA. We used linear regression modeling to test for associations between HuNoV antibody levels and sociodemographic factors, and with the human susceptibility rs601338 FUT2 secretor SNP and histo-blood group antigens (A/B/O)., Results: Of EMaBS participants, 76.6% were seropositive by age 1, rising to 94.5% by age 2 years. Seroprevalence in 1 year olds of the rural LaVIISWA survey was even higher (95%). In the birth cohort, 99% of seropositive 2 year olds had responses to multiple HuNoV genotypes. We identified associations between secretor status and genogroup GII antibody levels (GII.4 P = 3.1 × 10-52), as well as ABO and GI (GI.2 P = 2.1 × 10-12)., Conclusions: HuNoVs are highly prevalent in Ugandan children, indicating a substantial burden of diarrhea-associated morbidity with recurrent infections. Public health interventions, including vaccination, and increased surveillance are urgently needed.

BACKGROUND The aim of this study was to investigate the association between A, B, O, Rhesus (Rh)-positive and Rh-negative blood groups and breast cancer in a nationwide cohort of 3,944 patients in Turkey. MATERIAL AND METHODS A retrospective study included 3,944 patients diagnosed with breast cancer between 2004 and 2015 and with known blood type. Clinical and demographic patient data included age, sex, body mass index (BMI), menopausal status. The breast tumor type, size, grade, TNM stage, and the presence of lymph node and distant metastases were noted. Histopathology of the breast tumors had included routine detection of human epidermal growth factor receptor 2 (HER2) and estrogen receptor (ER) levels. RESULTS The 3,944 patients with breast cancer were blood group, type A, B, O, and Rh-positive or Rh-negative; the median age was 47.9 years (range, 18.2-89.6 years); 99.5% (3923/3,844) were women, and 0.5% (21/3944) were men. Patients with blood type 0 had a significantly smaller tumor size compared with patients with blood types A or B. There were no significant differences between blood groups and patient age, BMI, menopausal status, tumor histology, ER status, HER2 status, lymph node and distant metastasis. However, there was a significant difference in the prevalence of lobular breast cancer, levels of ER-positive tumor cells, and prevalence of cases with tumor metastases in Rh-positive patients compared with Rh-negative patients. CONCLUSIONS The findings of this retrospective study showed that the type, grade, stage, and hormonal status of breast cancer showed no significant associations with ABO blood grouping.

Conclusions: Antigens belonging to the Rh and Kell blood group systems are of major clinical significance because of their immunogenicity and the potential of their consequent antibodies to cause in vivo destruction of exogenous red blood cells (RBCs). Despite the wide-spread use of transfusion, there are sparse data on the prevalence of Rh and Kell system antigens and their ethnic variability in Nigeria. The objective of this study was to determine the prevalence of the five major Rh (D, C, c, E, e) and Kell (K) system antigens in Nigeria with the goal of understanding alloimmunization risk in transfusion recipients and improving transfusion safety through the availability of resources, such as antisera for extended RBC typing and antigen panels for alloantibody detection. A multi-ethnic cohort of 302 healthy Nigerian individuals was created to study RBC antigen prevalence. The antigen status of these individuals for Rh and K antigens was determined using commercially prepared antisera and conventional tube agglutination methods. The prevalence of the Rh antigens in the study cohort was found to be: D (92.7%), C (20.5%), c (97.7%), E (19.5%), and e (97.4%). Dce was the most common Rh phenotype (53.3%). The prevalence of K was 0 percent. For all antigens, there was no association between ethnicity and antigen prevalence. This study is the first to document the prevalence of the major Rh and K antigens in the Nigerian population, using a multi-ethnic cohort. Serologic testing demonstrates a zero prevalence of K antigen, which has never been described. C and E pose the higher risks of alloimmunization, hence showing a need for extended RBC typing and matching in at-risk blood recipients. This study demonstrates that phenotyping for major Rh and K antigens within the Nigerian population can potentially improve transfusion safety and prevent alloimmunization.

Introduction Studies show that rates of blood transfusion associated with general surgical laparoscopy are low. Currently, there are no national guidelines in the UK regarding blood group and antibody screening (G&S) for patients undergoing emergency laparoscopy. The aim of this study was to assess whether using G&S before emergency laparoscopic general surgery routinely is worthwhile by identifying rates of perioperative transfusion. Methods Data were collected retrospectively on all emergency laparoscopic procedures at a single district general hospital between January 2014 and 31 December 2016. Emergency laparoscopic general surgical cases were included and gynaecological cases excluded. Records were reviewed to ascertain whether G&S was performed, whether antibodies were detected and whether patients were transfused. Results A total of 562 emergency laparoscopic cases were performed. The median age was 28 years (range: 6-95 years). Laparoscopic appendicectomy (n=446), diagnostic laparoscopy (n=47) and laparoscopic cholecystectomy (n=25) were the most common procedures. Of the total patient cohort, 514 (91.5%) and 349 (70.1%) had a first and second G&S respectively while 30 (5.3%) had no G&S. Four patients (0.71%) had antibodies detected. One patient (0.18%) received a transfusion. This patient had undergone laparoscopic repair of a perforated duodenal ulcer and there was no major intraoperative haemorrhage but he was transfused perioperatively for chronic anaemia. Conclusions These results demonstrate a low rate of blood transfusion in emergency laparoscopic general surgery. The majority of these patients had a low risk of major intraoperative haemorrhage and we therefore argue that G&S was not warranted. We propose a more targeted approach to the requirement for preoperative G&S and the use of O negative blood in the event of acute haemorrhage from major vessel injury.

Background: The overall number of red blood cell (RBC) units distributed to hospitals throughout the world and in the United States has decreased lately. This study was performed to determine if the number of antigen-negative RBC units distributed to hospitals has followed this trend., Study Design and Methods: Stratified by ethnicity, data on total RBC distributions and antigen-negative RBC distributions from six large blood collectors in the United States were obtained from 2009 through 2016. An antigen-negative unit was defined as a unit with a specific RBC phenotype that had been specially ordered as such by a hospital., Results: Overall, 10,103,703 RBC units were distributed by these six blood collectors; 650,516 (6.4%) were distributed as antigen-negative units. While the overall number of RBCs distributed decreased by 27.2% between 2009 and 2016, the number of antigen-negative RBC distributions increased by 39.5%. In each year, the majority of the distributed antigen-negative RBCs were donated by whites. However, antigen-negative RBC units from black or African American donors were distributed in a disproportionately high fraction compared to the overall number of RBCs distributed from these donors. Most of the one through four antigen-negative RBCs were donated by whites. However, as antigen matching became more extensive, the proportion of units distributed from black or African American donors increased such that they were the predominant donors of five or more antigen-negative units., Conclusion: Blood collectors will need to be aware of the trend of increasing antigen-negative distributions despite decreased overall distributions., (© 2017 AABB.)

High-quality reagents to study and detect glycans with high specificity for research and clinical applications are severely lacking. Here, we structurally and functionally characterize several variable lymphocyte receptor (VLR)-based antibodies from lampreys immunized with O erythrocytes that specifically recognize the blood group H-trisaccharide type II antigen. Glycan microarray analysis and biophysical data reveal that these VLRs exhibit greater specificity for H-trisaccharide compared with the plant lectin UEA-1, which is widely used in blood typing. Among these antibodies, O13 exhibits superior specificity for H-trisaccharide, the basis for which is revealed by comparative analysis of high-resolution VLR:glycan crystal structures. Using a structure-guided approach, we designed an O13 mutant with further enhanced specificity for H-trisaccharide. These insights into glycan recognition by VLRs suggest that lampreys can produce highly specific glycan antibodies, and are a valuable resource for the production of next-generation glycan reagents for biological and biomedical research and as diagnostics and therapeutics., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Atomic force microscopy (AFM) offers complementary imaging modes that can provide morphological and structural details of red blood cells (RBCs), and characterize interactions between specific biomolecules and RBC surface antigen. This review describes the applications of AFM in determining RBC health by the observation of cell morphology, elasticity and surface roughness. Measurement of interaction forces between plasma proteins and antibodies against RBC surface antigen using the AFM also brought new information to the immunohaematology field. With constant improvisation of the AFM in resolution and imaging time, the reaction of RBC to changes in the physico-chemistry of its environment and the presence of RBC surface antigen specific-biomolecules is achievable., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A novel approach in antibody-free blood typing based on molecularly imprinted polymeric nanoparticles is described.

Background: Assessment of blood compatibility, typically by tube agglutination (TUBE) and hemolysis crossmatch or, less commonly, by blood typing and alloantibody screening, often is performed before blood transfusion in horses. In contrast, gel column (GEL) and immunochromatographic strip (STRIP) techniques are preferred for compatibility testing in dogs and cats., Objective: To determine the accuracy of novel and standard crossmatch and typing methods., Animals: Thirty-eight healthy horses, previously blood typed and alloantibody screened., Methods: TUBE and GEL crossmatches were performed on 146 different recipient-donor pairs with 56 incompatible TUBE crossmatches. Crossmatches were compared by nonparametric area under the curve of receiver operating characteristic (AUC-ROC) analyses. Horses also were blood typed by the novel immunochromatographic Ca typing STRIP., Results: Compared to TUBE crossmatch, GEL had excellent accuracy for agglutination (AUC-ROC = 0.903), but marginal accuracy for hemolysis (AUC-ROC = 0.639). Compared to macroscopic TUBE, microscopic TUBE had excellent accuracy for agglutination (AUC-ROC = 0.912). The predicted crossmatch compatibility based on blood type and alloantibody assay showed excellent accuracy compared to TUBE and GEL (AUC-ROC = 0.843 and 0.897, respectively). However, there were more recipient-donor pairs identified as incompatible by both TUBE and GEL than predicted by blood type and antibody screen, suggesting the presence of unidentified alloantibodies. A Ca typing STRIP exhibited 100% sensitivity and specificity for the 35 Ca+ and 3 Ca- horses tested., Conclusions and Clinical Relevance: Gel column crossmatch and Ca typing immunochromatographic strip are simple and accurate methods to evaluate clinical blood compatibility., (Copyright © 2016 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

A study was carried out to determine the prevalence of blood group antigen dog erythrocyte antigen (DEA) 1.1 in mixed breed dogs in rural Chinamhora, Zimbabwe. DEA 1.1 is clinically the most important canine blood group as it is the most antigenic blood type; hence, DEA 1.1 antibodies are capable of causing acute haemolytic, potentially life-threatening transfusion reactions. In this study, blood samples were collected from 100 dogs in Chinamhora, and blood typing was carried out using standardised DEA 1.1 typing strips with monoclonal anti-DEA 1.1 antibodies (Alvedia® LAB DEA 1.1 test kits). Polymerase chain reaction for detecting Babesia spp. antigen was carried out on 58 of the samples. Of the 100 dogs, 78% were DEA 1.1 positive and 22% were DEA 1.1 negative. A significantly (p = 0.02) higher proportion of females (90.5%) were DEA 1.1 positive than males (69.0%). The probability of sensitisation of recipient dogs following first-time transfusion of untyped or unmatched blood was 17.2%, and an approximately 3% (2.95%) probability of an acute haemolytic reaction following a second incompatible transfusion was found. Babesia spp. antigen was found in 6.9% of the samples. No significant relationship (χ2 = 0.56, p = 0.45) was found between DEA 1.1 positivity and Babesia spp. antigen presence. Despite a low probability of haemolysis after a second incompatibility transfusion, the risk remains present and should not be ignored. Hence, where possible, blood typing for DEA 1.1 is recommended. A survey of DEA 3, 4, 5 and 7 in various breeds is also recommended.

Background: A major cross-match gel tube test is available for use in dogs yet has not been clinically evaluated., Objectives: This study compared cross-match results obtained using the gel tube and the standard tube methods for canine samples., Methods: Study 1 included 107 canine sample donor-recipient pairings cross-match tested with the RapidVet-H method gel tube test and compared results with the standard tube method. Additionally, 120 pairings using pooled sera containing anti-canine erythrocyte antibody at various concentrations were tested with leftover blood from a hospital population to assess sensitivity and specificity of the gel tube method in comparison with the standard method., Results: The gel tube method had a good relative specificity of 96.1% in detecting lack of agglutination (compatibility) compared to the standard tube method. Agreement between the 2 methods was moderate. Nine of 107 pairings showed agglutination/incompatibility on either test, too few to allow reliable calculation of relative sensitivity. Fifty percent of the gel tube method results were difficult to interpret due to sample spreading in the reaction and/or negative control tubes., Conclusions: The RapidVet-H method agreed with the standard cross-match method on compatible samples, but detected incompatibility in some sample pairs that were compatible with the standard method. Evaluation using larger numbers of incompatible pairings is needed to assess diagnostic utility. The gel tube method results were difficult to categorize due to sample spreading. Weak agglutination reactions or other factors such as centrifuge model may be responsible., (© 2016 American Society for Veterinary Clinical Pathology.)

Background: The treatment of Plasmodium vivax infections requires the use of primaquine, which can lead to severe haemolysis in glucose-6-phosphate dehydrogenase (G6PD)-deficient individuals. However, most of the Latin American countries, which are still endemic for vivax malaria, lack information on the distribution of G6PD deficiency (G6PDd). No survey has been performed so far in French Guiana. Herein, 80 individuals of the French Guianan Noir Marron population were scrutinized for red cell surface antigens of six blood group systems (ABO, Rh, Kell, Kidd, Duffy and MNS) and G6PD genetic polymorphisms. First, the sub-Saharan origin of the red cell phenotypes was assessed in relation with the literature. Then, given that the main sub-Saharan G6PDd variants are expected to be encountered, only the G6PD sequences of exons 4, 5, 6 and 9 were screened. This work aims at appraising the G6PD gene variation in this population, and thus, contributing to the G6PD piecemeal information in Latin America., Results: Ninety-seven percent (97 %) of the red cells are Fy(a- b-), either D+ C- E- c+ e+ or D+ C+ E- c+ e+ and 44 % exhibited the Fya-/Jkb-/S- combined phenotype. Noteworthy is the detection of the G6PD(Val68Met) variant characterized by c.202G > A transition, G6PD(Asn126Asp) variant characterized by c.376A>G transition and G6PD(Asp181Val) variant characterized by c.542A>T transversion of the G6PD gene in 22.5 % of the sample, characteristic of the A(-(202)), A and Santamaria G6PDd variants, respectively., Conclusions: French Guianan Noir Marron population represents a pool of Rh-D antigen positive, Duffy-negative and G6PD-deficient erythrocytes, the latter accounting for one in every eight persons. The present study provides the first community-based estimation of the frequency of G6PDd polymorphisms in French Guiana. These results contribute to the G6PD genetic background information puzzle in Latin America.

Background: Common day case laparoscopic procedures are usually safe, with low rates of bleeding complications. At our trust, most patients undergo pre-operative group and save (G&S) for these procedures, at a cost of £18.39 per sample excluding laboratory staffing costs. Our aim was to assess if routine G&S is indicated., Methods: We performed a retrospective review of all patients who underwent laparoscopic cholecystectomy (LC), laparoscopic inguinal hernia repair (LIH) and diagnostic laparoscopy (DL) in our institution between April 2012 and March 2014. Patients were identified using hospital coding records. Transfusion department records were reviewed to see which patients had undergone pre-operative G&S or cross-match, and peri-operative transfusion., Results: Five hundred and thirty-two procedures were performed in 2 years: 293 LC, 123 LIH and 116 DL. G&S was performed in 256 (87 %; LC), 67 (54 %; LIH) and 88 (76 %; DL), respectively. Zero patients were transfused for bleeding complications. One patient was transfused following diagnostic laparoscopy to optimise pre-existing anaemia. The total cost of G&S over the study period was £7558., Conclusion: Blood transfusion rates for bleeding complications following laparoscopic day case surgery are 0 % in our unit. G&S samples for these procedures cost £7558 over 2 years. Abandoning pre-operative G&S for these patients appears to be clinically indicated and would lead to substantial financial savings.

A new platform of a paper-based analytical device (PAD) for simultaneous forward and reverse ABO blood group typing has been reported. This platform can overcome the discrepancy results as influenced by the individual haematocrit. The test and the control of non-haemagglutination on each channel were performed in parallel. The PAD was fabricated by printing six parallel channels with wax onto Whatman No. 4 filter paper. An LF1 blood separation membrane was used for the separation of plasma from whole blood for reverse grouping. The blood group was identified by haemagglutination of the corresponding antigen-antibody. For forward grouping, Anti-A, -B and -A,B were treated on the test line of PAD, and inactivated Anti-A, -B and -A,B were immobilized on the control line. For reverse grouping, 30% standard A-cells, B- and O- were added to the test channel after plasma separation, and O-cells were used as a control. Then, 0.9% normal saline (NSS) containing 1% Tween-20 was bi-functionally used for dilution of the blood sample and elution of the non-agglutinated RBCs within the channels. The distance of agglutinated RBCs in each test line was compared with the distance of non-agglutinated RBCs in the parallel control line. The forward and reverse patterns of blood groups A, B, AB and O were a barcode-like chart in which the results can be visually analysed. The PAD has excellent reproducibility when 10 replications of the A, B, AB or O blood groups were performed. The results of both forward and reverse grouping were highly correlated with conventional methods compared with the slide method and tube method, respectively (n = 76). Thus, this ABO typing PAD holds great potential for future applications in blood typing point-of-care testing., (Copyright © 2016 Elsevier B.V. All rights reserved.)