SG-APSIC1054: Sputnik-V postvaccination immunologic responses in nasal mucosa: A prospective cohort study in Kazakhstan

Objectives: Sputnik-V (Gam-COVID-Vac) is a recombinant adenoviral (rAdv) vector-based, COVID-19 vaccine now used in >70 countries. Mucosal immunity is thought to be important for protection against COVID-19. We did a prospective cohort study to assess Sputnik-V–elicited mucosal SARS-CoV-2 antibody responses. Methods: We divided 82 COVID-19–free participants into prior COVID-19 and no prior COVID-19 groups and followed them at day 21 after Sputnik-V dose 1′ (rAd5) and dose 2′ (rAd26). Nasopharyngeal swabs and blood were collected to perform SARS-CoV-2 diagnostic and immunologic assays. SARS-CoV-2 spike-specific IgG and IgA ELISAs were performed on both nasal swabs and blood. SARS-CoV-2 real-time RT-PCR testing was performed to exclude infectious influencing. Results: Nasal S-IgG levels increased 25-fold after dose 1′ (P < .001) and remained high after dose 2 in all participants. Prior COVID-19 exposure was associated with both elevated baseline mucosal IgG and IgA and higher postvaccination IgG, but not IgA, boost. Nasal IgA levels increased 16.5-fold after dose 1′ (P < .001) and remained high after dose 2' in all participants. Compared to dose 1′, Sputnik-V dose 2′ did not further increase either mucosal IgG levels (P = .626) or IgA levels (P = .609). Conclusions: A single dose of Sputnik-V boosted mucosal SARS-CoV-2 immunity. The effects of Sputnik-V dose 2′ on mucosal immunity were minimal. These findings indicate (1) that intramuscularly administered adenoviral vaccines enhance SARS-CoV-2 immunity via both systemic and mucosal routes and (2) that cost-effectiveness and the efficacy of Sputnik-V vaccination could be improved by adjusting the current prime-booster regimen and extending the 21-day interval between the doses. Trial registration: Registered on ClinicalTrials.gov (no. NCT04871841).

SG-APSIC1053: Detection of SARS-COV-2 in nasopharyngeal swags with MALDI-TOF MS and machine learning

Objectives: The widespread distribution of SARS-CoV-2 and its high contagiousness pose a challenge for researchers seeking to develop a rapid and cost-effective screening method to identify carriers of this virus. RT-PCR is considered the gold standard for detecting viral RNA in nasopharyngeal swabs, but it is time-consuming and requires constant changes in the primer composition due to the mutation of SARS-CoV-2 strains. We propose a method for the detection of SARS-CoV-2 in nasopharyngeal swabs using MALDI-TOF MS and machine learning. Methods: Nasopharyngeal swabs from patients with PCR-confirmed COVID-19 and control participants were tested (130 and 80 swabs, respectively) with MALDI-TOF MS MicroFlex LT using the HCCA matrix. MALDI spectra were preprocessed in R version 4.1.2 software with the MALDIquant R package using the workflow: sqrt transformation, wavelet smoothing, SNIP-based base removal, and PQN intensity calibration. Peaks were detected with MAD algorithms with following Peak alignment on the following parameters: minFreq 70% and tolerance 0.005. Machine learning was performed with the rtemis r package on GLM, random forest, and XGBoost models. Results: These models were characterized by specificity, sensitivity, and F1 score. GLM models (specificity 1 and sensitivity 0.5) showed a low F1 score of 0.71. However, the random forest and XGBoost models demonstrated sensitivity, specificity, and F1 score equaling 1. Conclusions: We propose a screening method for SARS-CoV-2 detection (sensitivity 1 and specificity 1). This methodology combines the analysis of nasopharyngeal swab samples using MALDI-TOF-MS with machine learning. It is suitable for screening patients with COVID-19 at the first stages of diagnosis. Random forest and XGBoost models demonstrated sensitivity, specificity, and F1 scores equaling 1.

Bacterial Co-Infections and Antimicrobial Resistance in Patients Hospitalized with Suspected or Confirmed COVID-19 Pneumonia in Kazakhstan.

Our study was carried out to characterize respiratory tract microbiota in patients with "COVID-like pneumonia" in Kazakhstan and analyze differences between COVID-19 positive and negative groups. Sputum samples were collected from hospitalized patients, ≥18 years old, in the three cities in Kazakhstan with the highest COVID-19 burden in July 2020. Isolates were identified by MALDI-TOF MS. Susceptibility testing was performed by disk diffusion. We used SPSS 26 and MedCalc 19 for statistical analysis. Among 209 patients with pneumonia, the median age was 62 years and 55% were male. RT-PCR-confirmed SARS-CoV-2 cases were found in 40% of patients, and 46% had a bacterial co-infection. Co-infection was not associated with SARS-CoV-2 RT-PCR test results, but antibiotic use was. The most frequent bacteria were Klebsiella pneumoniae (23%), Escherichia coli (12%), and Acinetobacter baumannii (11%). Notably, 68% of Klebsiella pneumoniae had phenotypic evidence of extended-spectrum beta-lactamases in disk diffusion assays, 87% of Acinetobacter baumannii exhibited resistance to beta-lactams, and >50% of E. coli strains had evidence of ESBL production and 64% were resistant to fluoroquinolones. Patients with a bacterial co-infection had a higher proportion of severe disease than those without a co-infection. The results reinforce the importance of using appropriate targeted antibiotics and effective infection control practices to prevent the spread of resistant nosocomial infections.

Mortality Risk Factors of Early Neonatal Sepsis During COVID-19 Pandemic.

Purpose: This study aimed to determine predisposing factors for negative outcome in infants with early neonatal sepsis during COVID-19. Patients and Methods: A prospective cohort study of 172 newborns up to 4 days diagnosed with neonatal sepsis was carried out in Karaganda (Kazakhstan). The microbiological examination was used to identify a causative agent of bloodstream infection. ELISA was performed to determine the total anti-SARS-CoV-2 antibodies. Gestational age, mode of delivery, birth weight, C-reactive protein and procalcitonin levels, comorbidities, type of pathogen, duration of hospitalization and mother's infection diseases were used for statistical analysis. Results: Mortality in infants with neonatal sepsis was 22% (38/172). Anti-SARS-CoV-2 antibodies were detected in 68.3% of the newborns. Culture-negative ELBW infants have a 5.3-fold higher risk of death (p<0.001). Low gestational age and a shorter period of hospitalization were statistically associated with fatality. CRP is generally higher in deceased children (p=0.002). Necrotizing enterocolitis (p<0.001), pneumonia (p=0.009) and anemia (p=0.016) were significantly associated with negative outcome. And, 31.4% of the infants with sepsis had positive blood cultures. The leading cause of sepsis in newborns was CoNS - 57%. Conclusion: During COVID-19 pandemic neonatal sepsis mortality was associated with low birth weight, gestational age, and comorbidities as in non-pandemic time. The relationship between COVID-19 in the mother and neonatal mortality was not found. However, anti-SARS-CoV-2 antibodies were detected in more than half of newborns.

Sputnik-V reactogenicity and immunogenicity in the blood and mucosa: a prospective cohort study.

Sputnik-V (Gam-COVID-Vac) is a heterologous, recombinant adenoviral (rAdv) vector-based, COVID-19 vaccine now used in > 70 countries. Yet there is a shortage of data on this vaccine's performance in diverse populations. Here, we performed a prospective cohort study to assess the reactogenicity and immunologic outcomes of Sputnik-V vaccination in Kazakhstan. COVID-19-free participants (n = 82 at baseline) were followed at day 21 after Sputnik-V dose 1 (rAd5) and dose 2 (rAd26). Self-reported local and systemic adverse events were captured using questionnaires. Blood and nasopharyngeal swabs were collected to perform SARS-CoV-2 diagnostic and immunologic assays. We observed that most of the reported adverse events were mild-to-moderate injection site or systemic reactions, no severe or potentially life-threatening conditions were reported, and dose 1 appeared to be more reactogenic than dose 2. The seroconversion rate was 97% post-dose 1, remaining the same post-dose 2. The proportion of participants with detectable virus neutralization was 83% post-dose 1, increasing to 98% post-dose 2, with the largest relative increase observed in participants without prior COVID-19 exposure. Dose 1 boosted nasal S-IgG and S-IgA, while the boosting effect of dose 2 on mucosal S-IgG, but not S-IgA, was only observed in subjects without prior COVID-19. Systemically, vaccination reduced serum levels of growth regulated oncogene (GRO), which correlated with an elevation in blood platelet count. Overall, Sputnik-V dose 1 elicited both blood and mucosal SARS-CoV-2 immunity, while the immune boosting effect of dose 2 was minimal. Thus, adjustments to the current vaccine dosing regimen are necessary to optimize immunization efficacy and cost-effectiveness. While Sputnik-V reactogenicity is similar to that of other COVID-19 vaccines, the induced alterations to the GRO/platelet axis warrant investigation of the vaccine's effects on systemic immunology.

High SARS-CoV-2 seroprevalence in Karaganda, Kazakhstan before the launch of COVID-19 vaccination.

COVID-19 exposure in Central Asia appears underestimated and SARS-CoV-2 seroprevalence data are urgently needed to inform ongoing vaccination efforts and other strategies to mitigate the regional pandemic. Here, in a pilot serologic study we assessed the prevalence of SARS-CoV-2 antibody-mediated immunity in a multi-ethnic cohort of public university employees in Karaganda, Kazakhstan. Asymptomatic subjects (n = 100) were recruited prior to their first COVID-19 vaccination. Questionnaires were administered to capture a range of demographic and clinical characteristics. Nasopharyngeal swabs were collected for SARS-CoV-2 RT-qPCR testing. Serological assays were performed to detect spike (S)-reactive IgG and IgA and to assess virus neutralization. Pre-pandemic samples were used to validate the assay positivity thresholds. S-IgG and -IgA seropositivity rates among SARS-CoV-2 PCR-negative participants (n = 100) were 42% (95% CI [32.2-52.3]) and 59% (95% CI [48.8-69.0]), respectively, and 64% (95% CI [53.4-73.1]) of the cohort tested positive for at least one of the antibodies. S-IgG titres correlated with virus neutralization activity, detectable in 49% of the tested subset with prior COVID-19 history. Serologically confirmed history of COVID-19 was associated with Kazakh ethnicity, but not with other ethnic minorities present in the cohort, and self-reported history of respiratory illness since March 2020. Overall, SARS-CoV-2 exposure in this cohort was ~15-fold higher compared to the reported all-time national and regional COVID-19 prevalence, consistent with recent studies of excess infection and death in Kazakhstan. Continuous serological surveillance provides important insights into COVID-19 transmission dynamics and may be used to better inform the regional public health response.