SARS-CoV-2 seroprevalence in Nova Scotia blood donors.

Background: SARS-CoV-2 seroprevalence monitors cumulative infection rates irrespective of case testing protocols. We aimed to describe Nova Scotia blood donor seroprevalence in relation to public health policy and reported data over the course of the COVID-19 pandemic (May 2020 to August 2022). Methods: Monthly random Nova Scotia blood donation samples (24,258 in total) were tested for SARS-CoV-2 infection antibodies (anti-nucleocapsid) from May 2020 to August 2022, and vaccination antibodies (anti-spike) from January 2021 to August 2022. Multivariable logistic regression for infection antibodies and vaccination antibodies separately with month, age, sex, and racialization identified independent predictors. The provincial nucleic acid amplification test (NAAT)-positive case rate over the pandemic was calculated from publicly available data. Results: Anti-N seroprevalence was 3.8% in January 2022, increasing to 50.8% in August 2022. The general population COVID-19 case rate was 3.5% in January 2022, increasing to 12.5% in August 2022. The percentage of NAAT-positive samples in public health laboratories increased from 1% in November 2021 to a peak of 30.7% in April 2022 with decreasing numbers of tests performed. Higher proportions of younger donors as well as Black, Indigenous, and racialized blood donors were more likely to have infection antibodies (p < 0.01). Vaccination antibodies increased to 100% over 2021, initially in older donors (60+ years), and followed by progressively younger age groups. Conclusions: SARS-CoV-2 infection rates were relatively low in Nova Scotia until the more contagious Omicron variant dominated, after which about half of Nova Scotia donors had been infected despite most adults being vaccinated (although severity was much lower in vaccinated individuals). Most COVID-19 cases were detected by NAAT until Omicron arrived. When NAAT testing priorities focused on high-risk individuals, infection rates were better reflected by seroprevalence.

Historique: La séroprévalence du SRAS-CoV-2 permet de surveiller les taux d'infection cumulatifs, quels que soient les protocoles de dépistage des cas. Les chercheurs voulaient décrire la séroprévalence chez les donneurs de sang néo-écossais par rapport aux politiques sanitaires et aux données déclarées tout au long de la pandémie de COVID-19 (mai 2020 à août 2022). Méthodologie: Les chercheurs ont mesuré les anticorps à l'infection par le SRAS-CoV-2 (antinucléocapsidiques) de manière aléatoire tous les mois dans des échantillons de dons de sang de la Nouvelle-Écosse (pour un total de 24 258) entre mai 2020 et août 2022, de même que les anticorps aux vaccins (antispiculaires) (entre janvier 2021 et août 2022). La régression logistique multivariable effectuée séparément pour les anticorps contre l'infection et les anticorps aux vaccins, par rapport au mois, à l'âge, au genre et à la race, ont permis d'établir les prédicteurs indépendants. Les chercheurs ont calculé le taux de cas provinciaux positifs aux tests d'amplification des acides nucléiques (TAAN) à partir des données publiques tout au long de la pandémie. Résultats: La séroprévalence anti-N, qui s'élevait à 3,8 % en janvier 2022, était passée à 50,8 % en août 2022. Le taux de cas de COVID-19 dans la population générale se situait à 3,5 % en janvier 2022 et était monté à 12,5 % en août 2022. Le pourcentage d'échantillons positifs au TAAN dans les laboratoires de santé publique est passé de 1 % en novembre 2021 à un pic de 30,7 % en avril 2022, conjointement à une diminution du nombre de tests effectués. De plus fortes proportions de donneurs plus jeunes et de donneurs noirs, autochtones et racisés étaient susceptibles de posséder des anticorps contre l'infection (p < 0,01). Les anticorps attribuables à la vaccination ont atteint 100 % en 2021, d'abord chez les donneurs plus âgés (de plus de 60 ans), puis dans les tranches d'âges progressivement plus jeunes. Conclusions: Les taux d'infection par le SRAS-CoV-2 sont demeurés relativement faibles en Nouvelle-Écosse jusqu'à la domination du variant Omicron plus contagieux, puis environ la moitié des donneurs néo-écossais ont été infectés, même si la plupart des adultes étaient vaccinés (la gravité de la maladie était toutefois beaucoup plus faible chez les personnes vaccinées). La plupart des cas de COVID-19 ont été dépistés par TAAN jusqu'à l'apparition du variant Omicron. Lorsque les priorités de dépistage par TAAN ont été limitées aux personnes à haut risque, les taux d'infection étaient mieux reflétés par la séroprévalence.

Post-Reconstitution Hemostatic Stability Profiles of Canadian and German Freeze-Dried Plasma.

Despite the importance of the hemostatic properties of reconstituted freeze-dried plasma (FDP) for trauma resuscitation, few studies have been conducted to determine its post-reconstitution hemostatic stability. This study aimed to assess the short- (≤24 h) and long-term (≥168 h) hemostatic stabilities of Canadian and German freeze-dried plasma (CFDP and LyoPlas) after reconstitution and storage under different conditions. Post-reconstitution hemostatic profiles were determined using rotational thromboelastometry (ROTEM) and a Stago analyzer, as both are widely used as standard methods for assessing the quality of plasma. When compared to the initial reconstituted CFDP, there were no changes in ROTEM measurements for INTEM maximum clot firmness (MCF), EXTEM clotting time (CT) and MCF, and Stago measurements for prothrombin time (PT), partial thromboplastin time (PTT), D-dimer concentration, plasminogen, and protein C activities after storage at 4 °C for 24 h and room temperature (RT) (22-25 °C) for 4 h. However, an increase in INTEM CT and decreases in fibrinogen concentration, factors V and VIII, and protein S activities were observed after storage at 4 °C for 24 h, while an increase in factor V and decreases in antithrombin and protein S activities were seen after storage at RT for 4 h. Evaluation of the long-term stability of reconstituted LyoPlas showed decreased stability in both global and specific hemostatic profiles with increasing storage temperatures, particularly at 35 °C, where progressive changes in CT and MCF, PT, PTT, fibrinogen concentration, factor V, antithrombin, protein C, and protein S activities were seen even after storage for 4 h. We confirmed the short-term stability of CFDP in global hemostatic properties after reconstitution and storage at RT, consistent with the shelf life of reconstituted LyoPlas. The long-term stability analyses suggest that the post-reconstitution hemostatic stability of FDP products would decrease over time with increasing storage temperature, with a significant loss of hemostatic functions at 35 °C compared to 22 °C or below. Therefore, the shelf life of reconstituted FDP should be recommended according to the storage temperature.

The operational impact of introducing cold stored platelets.

BACKGROUND: Cold stored platelets (CSP) undergo physical changes that make them better at initiating a clot. While cold stored platelets are superior for reducing bleeding in actively bleeding patients, room temperature platelets (RTP) are better for increasing platelet count in patients requiring a prophylactic transfusion. However, whether the overhead required to maintain a dual platelet inventory of both RTP and CSP could be compensated by reduced platelet wastage resulting from the longer shelf life of CSP has not been determined. STUDY DESIGN AND METHODS: A simulation model of a regional blood supply was built, with focus on the operations of a case hospital. Two scenarios were considered: "No-CSP," in which the hospital issues only RTP, and "CSP," in which the hospital issues both RTP and CSP Within the CSP scenario, conditions were tested under which the hospital receives only RTP and converts some to cold stored platelets and a second strategy where the hospital receives CSP from the regional supplier in addition to converting RTP. RESULTS: A centralized supply of CSP is necessary since on-site conversion is limited by platelet age. Product shortages decrease with increased CSP inventory, but CSP wastage increases. It was also determined that, because relatively few RTP units can be converted on-site, RTP wastage is not significantly decreased with the introduction of CSP. CONCLUSION: Given the clinical benefits for treatment of trauma, CSP is a desirable addition to a blood formulary. However, it is unlikely that significant reductions in RTP wastage will occur because of the introduction of CSP.

The evolution of SARS-CoV-2 seroprevalence in Canada: a time-series study, 2020-2023.

BACKGROUND: During the first year of the COVID-19 pandemic, the proportion of reported cases of COVID-19 among Canadians was under 6%. Although high vaccine coverage was achieved in Canada by fall 2021, the Omicron variant caused unprecedented numbers of infections, overwhelming testing capacity and making it difficult to quantify the trajectory of population immunity. METHODS: Using a time-series approach and data from more than 900 000 samples collected by 7 research studies collaborating with the COVID-19 Immunity Task Force (CITF), we estimated trends in SARS-CoV-2 seroprevalence owing to infection and vaccination for the Canadian population over 3 intervals: prevaccination (March to November 2020), vaccine roll-out (December 2020 to November 2021), and the arrival of the Omicron variant (December 2021 to March 2023). We also estimated seroprevalence by geographical region and age. RESULTS: By November 2021, 9.0% (95% credible interval [CrI] 7.3%-11%) of people in Canada had humoral immunity to SARS-CoV-2 from an infection. Seroprevalence increased rapidly after the arrival of the Omicron variant - by Mar. 15, 2023, 76% (95% CrI 74%-79%) of the population had detectable antibodies from infections. The rapid rise in infection-induced antibodies occurred across Canada and was most pronounced in younger age groups and in the Western provinces: Manitoba, Saskatchewan, Alberta and British Columbia. INTERPRETATION: Data up to March 2023 indicate that most people in Canada had acquired antibodies against SARS-CoV-2 through natural infection and vaccination. However, given variations in population seropositivity by age and geography, the potential for waning antibody levels, and new variants that may escape immunity, public health policy and clinical decisions should be tailored to local patterns of population immunity.

Cross-Canada Variability in Blood Donor SARS-CoV-2 Seroprevalence by Social Determinants of Health.

We compared the seroprevalence of SARS-CoV-2 anti-nucleocapsid antibodies in blood donors across Canadian regions in 2021. The seroprevalence was the highest in Alberta and the Prairies, and it was so low in Atlantic Canada that few correlates were observed. Being male and of young age were predictive of seropositivity. Racialization was associated with higher seroprevalence in British Columbia and Ontario but not in Alberta and the Prairies. Living in a materially deprived neighborhood predicted higher seroprevalence, but it was more linear across quintiles in Alberta and the Prairies, whereas in British Columbia and Ontario, the most affluent 60% were similarly low and the most deprived 40% similarly elevated. Living in a more socially deprived neighborhood (more single individuals and one parent families) was associated with lower seroprevalence in British Columbia and Ontario but not in Alberta and the Prairies. These data show striking variability in SARS-CoV-2 seroprevalence across regions by social determinants of health. IMPORTANCE Canadian blood donors are a healthy adult population that shows clear disparities associated with racialization and material deprivation. This underscores the pervasiveness of the socioeconomic gradient on SARS-CoV-2 infections in Canada. We identify regional differences in the relationship between SARS-CoV-2 seroprevalence and social determinants of health. Cross-Canada studies, such as ours, are rare because health information is under provincial jurisdiction and is not available in sufficient detail in national data sets, whereas other national seroprevalence studies have insufficient sample sizes for regional comparisons. Ours is the largest seroprevalence study in Canada. An important strength of our study is the interpretation input from a public health team that represented multiple Canadian provinces. Our blood donor seroprevalence study has informed Canadian public health policy at national and provincial levels since the start of the SARS-CoV-2 pandemic.

SARS-CoV-2 Seroprevalence among Canadian Blood Donors: The Advance of Omicron.

With the emergence of the SARS-CoV-2 Omicron variant in late 2021, Canadian public health case/contact testing was scaled back due to high infection rates with milder symptoms in a highly vaccinated population. We monitored the seroprevalence of SARS-CoV-2 nucleocapsid (anti-N) and spike protein (anti-S) antibodies in blood donors across Canada from September 2021 to June 2022 in 202,123 randomly selected samples. Multivariable logistic regression of anti-N positivity with month, age, sex, racialization, region, material and social deprivation (based on postal code) identified as independent predictors. Piece-wise logistic regression analysed the association between anti-S concentration and month, and anti-N/anti-S positivity. Infection-related seroprevalence (anti-N positive) was 4.38% (95% CI: 3.96, 4.81) in September reaching 50.70% (50.15, 52.16) in June; nearly 100% were anti-S positive throughout. Anti-N positivity was associated with younger age, male sex, the Alberta and Prairies regions, greater material deprivation and less social deprivation (p < 0.001). Anti-S concentration was high initially (3306 U/mL, IQR 4280 U/mL), increased to (13,659 U/mL, IQR 28,224 U/mL) by June (p < 0.001), following the pattern of deployment of the third and fourth vaccine doses and was higher in those that were anti-N positive (p < 0.001). Despite already high vaccination-related seroprevalence, infection-related seroprevalence increased dramatically with the emergence of the Omicron SARS-CoV-2 variant.

Changing Patterns of SARS-CoV-2 Seroprevalence among Canadian Blood Donors during the Vaccine Era.

We monitored the seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid (anti-N; proxy of natural infection) and spike protein (anti-S; proxy for humoral immunity) antibodies in blood donors across Canada from January to November 2021. The first and second doses of vaccine were deployed over this time. Anti-N seroprevalence remained low overall (about 5% or lower) from January to November but was higher in racialized groups, younger age groups, and those living in materially deprived neighborhoods. Anti-S seroprevalence corresponded with the roll out of vaccines across the country, increasing in April in older donors and then progressively to younger age groups consistent with vaccination policies targeting oldest to youngest. By November, close to 100% of blood donors were positive for anti-S. Anti-S concentrations peaked by July and began waning by September to November particularly in older donors. These data have informed national and provincial public health policy in Canada throughout vaccination rollout. IMPORTANCE Throughout the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, our blood donor seroprevalence study has informed Canadian public health policy at national and provincial levels. We describe the only continuously running national seroprevalence study in Canada, which spans the full length of the pandemic and per capita is one of the largest programs in the world. The benefit of seroprevalence studies is that they identify a broad range of asymptomatic and symptomatic infection histories that may not be identified with active SARS-CoV-2 nucleic acid testing programs or when case definitions change. As vaccination was deployed in Canada, we estimated the proportion of donors with vaccine-related antibodies and developed population-level estimates of SARS-CoV-2 spike antibody concentrations. Monthly predictive mathematical models and our results engaged public health organizations in new ways. In the future, we intend to continue to expand on these interactions with provincial and national public health teams.

SARS-CoV-2 seroprevalence among blood donors after the first COVID-19 wave in Canada.

BACKGROUND: Case detection underestimates the burden of the COVID-19 pandemic. Following the first COVID-19 wave, we estimated the seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) among blood donors across Canada. STUDY DESIGN AND METHODS: This serial cross-sectional study was conducted between May 9 and July 21, 2020 from blood donors donating at all Canadian Blood Services locations. We used the Abbott Architect assay to detect SARS-CoV-2 IgG antibodies from retention plasma. Seroprevalence was standardized to population-level demographics and assay characteristics were adjusted using the Rogan-Gladen equation. Results were stratified by region, age, ethnicity, ABO groups, and quantiles of material and social deprivation indices. Temporal trends were evaluated at 2-week intervals. Univariate and multivariate logistic regression compared SARS-CoV-2 reactive to non-reactive donors by sociodemographic variables. RESULTS: Overall 552/74642 donors, had detectable antibodies, adjusted seroprevalence was 7.0/1000 donors (95% CI; 6.3, 7.6). Prevalence was differential by geography, Ontario had the highest rate, 8.8/1000 donors (7.8, 9.8), compared to the Atlantic region 4.5/1000 donors (2.6, 6.4); adjusted odds ratio (aOR) 2.2 (1.5, 3.3). Donors that self-identified as an ethnic minority were more likely than white donors to be sero-reactive aOR 1.5 (1.2, 1.9). No temporal trends were observed. DISCUSSION: Worldwide, blood services have leveraged their operational capacity to inform public health. While >99% of Canadians did not show humoral evidence of past infection, we found regional variability and disparities by ethnicity. Seroprevalence studies will continue to play a pivotal role in evaluating public health policies by identifying trends and monitor disparities.

Improving Time to Antibiotics for Pediatric Oncology Patients With Fever and Suspected Neutropenia by Applying Lean Principles.

BACKGROUND: Fever in the setting of neutropenia is a potentially life-threatening complication of cancer treatment. A time of less than 60 minutes from presentation to antibiotic administration is therefore recommended. OBJECTIVE: To use Lean Six Sigma methodology, a quality improvement initiative, to improve time to antibiotics (TTA) for children with chemotherapy-induced febrile neutropenia presenting to the emergency department. METHODS: Lean Six Sigma is a quality improvement method that engages all impacted stakeholders and focuses on streamlining the process by removing process wastes. Stakeholders identified multiple process wastes in an in-depth study of 49 fever episodes in patients attending a tertiary care pediatric hospital, including patients waiting to be registered, waiting for laboratory technicians, delay in accessing central venous access device, waiting for absolute neutrophil count, and delayed antibiotics orders. We implemented multiple solutions: engaging patients in the process through predischarge tours of the emergency department, home application of topical anesthetic, nurse-initiated pathway, early access of central venous access device for all blood work, and planned antibiotic administration no later than 45 minutes after triage. We prospectively determined the impact of these interventions on TTA. RESULTS: The TTA significantly improved to a median of 59 minutes (interquartile range, 38.5-77.5 minutes) compared with the baseline of 99 minutes (interquartile range, 72.0-132.0 minutes; P < 0.0001). CONCLUSIONS: Lean methodology effectively identifies barriers and provides solutions to remove barriers and improve administration of antibiotics in febrile oncology patients. These can be widely applied, including in smaller institutions with minimal increased utilization of resources.

Supernatant reduction of stored gamma-irradiated red blood cells minimizes potentially harmful substances present in transfusion aliquots for neonates.

BACKGROUND: In neonate transfusion, the use of a dedicated red blood cell (RBC) unit decreases donor exposure. A separate safety measure involves gamma irradiation of the RBCs to abrogate the possibility of transfusion-associated graft-versus-host disease. However, in combination, storage of gamma-irradiated RBCs leads to accumulation of potentially harmful substances in the supernatant. STUDY DESIGN AND METHODS: For this study, RBCs were pooled and split into three study arms. Centrifugation or gravity was used to pack RBCs of matched units thereby reducing the amount of supernatant that would be present in neonate transfusion aliquots; these were compared to matched control units. Supernatant measurements of potassium, hemoglobin (Hb), RBC microvesicle (RMV) content, and mannitol were made in aliquots prepared weekly up to 21 days after gamma irradiation. RBC morphology and osmotic fragility were also assessed to determine if supernatant reduction methods affected the storage lesion. RESULTS: Potassium and mannitol were significantly decreased in transfusion aliquots prepared with either of the supernatant reduction methods. On Day 21, potassium levels from supernatant-reduced aliquots were below those of Day 7 control aliquots. A decrease in free Hb was only detected on Day 21 in centrifuged aliquots. RMVs were significantly reduced in centrifuged aliquots and significantly increased in gravity-settled aliquots. The only measurable effect on storage lesion was a small increase in osmotic fragility of the RBCs subjected to supernatant reduction. CONCLUSION: Supernatant reduction by centrifugation effectively reduces potassium, mannitol, and RMVs in aliquots from gamma-irradiated RBCs stored up to 21 days.

Effect on gene expression of three allelic variants in GATA motifs of ABO, RHD, and RHCE regulatory elements.

BACKGROUND: Only a few genetic variants have been reported in regulatory elements of blood group genes. Most of them affect GATA motifs, binding sites for the GATA-1 transcription factor. STUDY DESIGN AND METHODS: Samples from two patients and one donor with unusual or discrepant serology for ABO, RhD, and RhCE antigens were analyzed by DNA sequencing. Analyzed regions included the coding sequence and portions of regulatory elements. The effect of some variants on gene expression was evaluated in reporter gene assays. RESULTS: Three new alleles were identified. Their key variants are located in the ABO Intron 1 enhancer, the RHD proximal promoter, and the RHCE proximal promoter. IVS1 + 5859A was found in an African American patient with a group O forward type and a group B reverse type. 5'UTR-115C was the only RHD variant sequence found in a mixed-race black and Caucasian prenatal patient showing mixed-field agglutination with anti-D. 5'UTR-83T was found in several black donors and patients in the context of the genetically related RHCE*ceBI and RHCE*ceSM alleles. Reporter assays of promoter constructs including 5'UTR-115C or 5'UTR-83T showed a significant reduction in RH gene expression. CONCLUSION: Three new alleles in the ABO, RHD, and RHCE genes consist of single-nucleotide changes within GATA motifs, emphasizing the key role of GATA transcription factors in the expression of blood group genes.

FISH identifies a KAT6A/CREBBP fusion caused by a cryptic insertional t(8;16) in a case of spontaneously remitting congenital acute myeloid leukemia with a normal karyotype.

Cytogenetics can inform risk stratification in pediatric acute myeloid leukemia (AML). We describe the first case of a newborn with leukemia cutis found to have AML harboring a cryptic insertional t(8;16)(p11.2;p13.3) with associated KAT6A/CREBBP fusion identified exclusively by fluorescence in situ hybridization (FISH). Expectant management resulted in spontaneous leukemia resolution. The identification of t(8;16)(p11.2;p13.3) may serve as a biomarker for spontaneous remission in congenital AML. FISH for this translocation is warranted in congenital AML with a normal karyotype, and patients with KAT6A/CREBBP fusion should be conservatively managed. While 50% of spontaneously remitting congenital AML with t(8;16)(p11.2;p13.3) may recur, high salvage rates are attained with standard therapy.

Study of Glycine and Folic Acid Supplementation to Ameliorate Transfusion Dependence in Congenital SLC25A38 Mutated Sideroblastic Anemia.

Congenital sideroblastic anemia (CSA) is a hematological disorder characterized by the presence of ringed sideroblasts in bone marrow erythroid precursors. Mutations in the erythroid-specific glycine mitochondrial transporter gene SLC25A38 have been found in a subset of patients with transfusion-dependent congenital CSA. Further studies in a zebrafish model identified a promising ameliorative strategy with combined supplementation with glycine and folate. We tested this combination in three individuals with SLC25A38 CSA, with a primary objective to decrease red blood cell transfusion requirements. No significant impact was observed on transfusion requirements or any hematologic parameters.

Congenital methemoglobinaemia due to Hb F-M-Fort Ripley in a preterm newborn.

Methemoglobinaemia is a rare cause of cyanosis in newborns. Congenital methemoglobinaemias due to M haemoglobin or deficiency of cytochrome b5 reductase are even rarer. We present a case of congenital methemoglobinaemia presenting at birth in a preterm infant. A baby boy born at 29 weeks and 3 days of gestation had persistent central cyanosis immediately after delivery, not attributable to a respiratory or cardiac pathology. Laboratory methemoglobin levels were not diagnostic. Cytochrome b5 reductase levels were normal and a newborn screen was unable to pick up any abnormal variants of fetal haemoglobin. Genetic testing showed a γ globin gene mutation resulting in the M haemoglobin, called Hb F-M-Fort Ripley. The baby had no apparent cyanosis at a corrected gestational age of 42 weeks. Although rare, congenital methaemoglobin aemia should be considered in the differential in a preterm with central cyanosis and investigated with genetic testing for γ globin chain mutations if other laboratory tests are non-conclusive.

Exit competencies in pathology and laboratory medicine for graduating medical students: the Canadian approach.

Physicians in every medical and surgical field must be able to use pathology concepts and skills in their practice: for example, they must order and interpret the correct laboratory tests, they must use their understanding of pathogenesis to diagnose and treat, and they must work with the laboratory to care for their patients. These important concepts and skills may be ignored by medical schools and even national/international organizations setting graduation expectations for medical students. There is an evolving international consensus about the importance of exit competencies for medical school graduates, which define the measurable or observable behaviors each graduate must be able to demonstrate. The Canadian Association of Pathologists (CAP) Education Group set out to establish the basic competencies in pathology and laboratory medicine which should be expected of every medical graduate: not competencies for pathologists, but for medical graduates who intend to enter any residency program. We defined 4 targets for pathology and laboratory medicine exit competencies: that they represent only measurable behaviors, that they be clinically focused, that they be generalizable to every medical graduate, and that the final competency document be user-friendly. A set of competencies was developed iteratively and underwent final revision at the 2012 CAP annual meeting. These competencies were subsequently endorsed by the CAP executive and the Canadian Leadership Council on Laboratory Medicine. This clinically focused consensus document provides the first comprehensive list of exit competencies in pathology and laboratory medicine for undergraduate medical education.