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Clin Transl Immunology ; 11(3): e1380, 2022.
Article in English | MEDLINE | ID: covidwho-1750347


Objectives: Antibody testing against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been instrumental in detecting previous exposures and analyzing vaccine-elicited immune responses. Here, we describe a scalable solution to detect and quantify SARS-CoV-2 antibodies, discriminate between natural infection- and vaccination-induced responses, and assess antibody-mediated inhibition of the spike-angiotensin converting enzyme 2 (ACE2) interaction. Methods: We developed methods and reagents to detect SARS-CoV-2 antibodies by enzyme-linked immunosorbent assay (ELISA). The main assays focus on the parallel detection of immunoglobulin (Ig)Gs against the spike trimer, its receptor binding domain (RBD) and nucleocapsid (N). We automated a surrogate neutralisation (sn)ELISA that measures inhibition of ACE2-spike or -RBD interactions by antibodies. The assays were calibrated to a World Health Organization reference standard. Results: Our single-point IgG-based ELISAs accurately distinguished non-infected and infected individuals. For seroprevalence assessment (in a non-vaccinated cohort), classifying a sample as positive if antibodies were detected for ≥ 2 of the 3 antigens provided the highest specificity. In vaccinated cohorts, increases in anti-spike and -RBD (but not -N) antibodies are observed. We present detailed protocols for serum/plasma or dried blood spots analysis performed manually and on automated platforms. The snELISA can be performed automatically at single points, increasing its scalability. Conclusions: Measuring antibodies to three viral antigens and identify neutralising antibodies capable of disrupting spike-ACE2 interactions in high-throughput enables large-scale analyses of humoral immune responses to SARS-CoV-2 infection and vaccination. The reagents are available to enable scaling up of standardised serological assays, permitting inter-laboratory data comparison and aggregation.

JAMA Netw Open ; 5(2): e2146798, 2022 02 01.
Article in English | MEDLINE | ID: covidwho-1694847


Importance: The incidence of infection during SARS-CoV-2 viral waves, the factors associated with infection, and the durability of antibody responses to infection among Canadian adults remain undocumented. Objective: To assess the cumulative incidence of SARS-CoV-2 infection during the first 2 viral waves in Canada by measuring seropositivity among adults. Design, Setting, and Participants: The Action to Beat Coronavirus study conducted 2 rounds of an online survey about COVID-19 experience and analyzed immunoglobulin G levels based on participant-collected dried blood spots (DBS) to assess the cumulative incidence of SARS-CoV-2 infection during the first and second viral waves in Canada. A sample of 19 994 Canadian adults (aged ≥18 years) was recruited from established members of the Angus Reid Forum, a public polling organization. The study comprised 2 phases (phase 1 from May 1 to September 30, 2020, and phase 2 from December 1, 2020, to March 31, 2021) that generally corresponded to the first (April 1 to July 31, 2020) and second (October 1, 2020, to March 1, 2021) viral waves. Main Outcomes and Measures: SARS-CoV-2 immunoglobulin G seropositivity (using a chemiluminescence assay) by major geographic and demographic variables and correlation with COVID-19 symptom reporting. Results: Among 19 994 adults who completed the online questionnaire in phase 1, the mean (SD) age was 50.9 (15.4) years, and 10 522 participants (51.9%) were female; 2948 participants (14.5%) had self-identified racial and ethnic minority group status, and 1578 participants (8.2%) were self-identified Indigenous Canadians. Among participants in phase 1, 8967 had DBS testing. In phase 2, 14 621 adults completed online questionnaires, and 7102 of those had DBS testing. Of 19 994 adults who completed the online survey in phase 1, fewer had an educational level of some college or less (4747 individuals [33.1%]) compared with the general population in Canada (45.0%). Survey respondents were otherwise representative of the general population, including in prevalence of known risk factors associated with SARS-CoV-2 infection. The cumulative incidence of SARS-CoV-2 infection among unvaccinated adults increased from 1.9% in phase 1 to 6.5% in phase 2. The seropositivity pattern was demographically and geographically heterogeneous during phase 1 but more homogeneous by phase 2 (with a cumulative incidence ranging from 6.4% to 7.0% in most regions). The exception was the Atlantic region, in which cumulative incidence reached only 3.3% (odds ratio [OR] vs Ontario, 0.46; 95% CI, 0.21-1.02). A total of 47 of 188 adults (25.3%) reporting COVID-19 symptoms during phase 2 were seropositive, and the OR of seropositivity for COVID-19 symptoms was 6.15 (95% CI, 2.02-18.69). In phase 2, 94 of 444 seropositive adults (22.2%) reported having no symptoms. Of 134 seropositive adults in phase 1 who were retested in phase 2, 111 individuals (81.8%) remained seropositive. Participants who had a history of diabetes (OR, 0.58; 95% CI, 0.38-0.90) had lower odds of having detectable antibodies in phase 2. Conclusions and Relevance: The Action to Beat Coronavirus study found that the incidence of SARS-CoV-2 infection in Canada was modest until March 2021, and this incidence was lower than the levels of population immunity required to substantially reduce transmission of the virus. Ongoing vaccination efforts remain central to reducing viral transmission and mortality. Assessment of future infection-induced and vaccine-induced immunity is practicable through the use of serial online surveys and participant-collected DBS.

COVID-19 Serological Testing/statistics & numerical data , COVID-19/epidemiology , Immunoglobulin G/blood , Adolescent , Adult , Aged , COVID-19/immunology , Canada/epidemiology , Female , Humans , Incidence , Male , Middle Aged , Pandemics , SARS-CoV-2 , Surveys and Questionnaires
EuropePMC; 2021.
Preprint in English | EuropePMC | ID: ppcovidwho-322524


Background: The prevalence of infection in Canada’s COVID-19 viral waves, the predictors of infection, and the durability of antibody responses to infection remain undocumented.Methods: We organized serial online surveys of a representative group of Canadian adults about their COVID experience in the first (n=19 994;April-July 2020) and second viral wave (n=14 621;October 2020-March 2021). We paired these with IgG analysis of SARS-CoV-2 seroprevalence in self-collected dried blood spots after the first (n=8967) and second (n=7102) waves.Findings: Canada’s cumulative seroprevalence of SARS-CoV-2 among unvaccinated adults rose from ~2% after the first wave to 7% after the second. The seropositivity pattern was heterogeneous demographically and geographically during the first wave, but more homogeneous by the second (except in the four Atlantic Provinces, cumulative seroprevalence ~3%). Seroprevalence among visible minorities rose sharply from about 2% to >8% from the first to second wave. About a quarter of those reporting COVID symptoms during the second wave were seropositive, and in both waves the odds ratio (OR) of seropositivity for COVID symptoms exceeded six. About one-fifth of seropositives reported no symptoms. Of 134 seropositive adults in the first wave who were retested after the second, 83% (111) remained seropositive at least seven months later. Current smokers and people with a history of diabetes had lower ORs of infection. We calculated the absolute numbers of seropositive adults nationwide, which nearly quadrupled from 0.57 million to 1.90 million, with the largest increases among older adults. Infection fatality rates fell from 3.7 to 2.6/1000 infections, most notably at older ages.Interpretation: Canada’s COVID pandemic grew substantially between the first and second viral waves. Home-based DBS collection offers a practicable way to document evolving demographic and geographic patterns and to assess the levels and durability of population immunity, including from SARS-CoV-2 vaccination.Funding: Pfizer Global Medical, Unity Health Foundation, and the Canadian COVID-19 Immunity Task Force. Declaration of Interest: None to declare. Ethical Approval: The Ab-C study was approved by the Unity Health Toronto Ethics Review Board.