Accuracy of interferon gamma release assays for the COVID-19 immunity assessment.

Emerging evidence suggests that T-cells play a significant role in COVID-19 immunity both in the context of natural infection and vaccination. Easy to use IGRA assays including QFN SARS are considered attractive alternatives to more "traditional" but laborious methods for detection of SARS-CoV-2-specific T-cell responses. In our Letter we are proposing explanations to an apparently lower than expected T-cell responses (44 % reactive individuals) reported by Krüttgen et al in a small cohort of healthy double vaccinated individuals. These results could have been affected by reporting raw optical density values instead of calculated Interferon-É£ concentrations which is supported by unexpectedly low mitogen responses in healthy individuals. This study highlights an importance of adhering to good laboratory practice principles as well as overall importance of accurate T-cell immunity assessment using IGRA assays.

Preliminary Evaluation of QuantiFERON SARS-CoV-2 and QIAreach Anti-SARS-CoV-2 Total Test in Recently Vaccinated Individuals.

INTRODUCTION: There is an increasing body of evidence surrounding the importance of a T cell-mediated response to SARS-CoV-2 infection and after COVID-19 vaccination. In this internal feasibility study, we evaluated both the total antibody (IgA, IgM, and IgG) and T cell responses in a cohort of COVID-19 convalescents and vaccinated individuals. METHODS: Whole blood specimens were collected weekly from 12 subjects at different time points within/after the COVID-19 mRNA vaccination regimen, and from 4 PCR-confirmed convalescent donors to measure durability of humoral and cell-mediated immune response. T cell and antibody responses were evaluated via the QuantiFERON SARS-CoV-2 research use only (QFN SARS-CoV-2) assay which is an interferon gamma release assay (IGRA) and QIAreach Anti-SARS-CoV-2 total (Anti-CoV-2) test, respectively. RESULTS: In a cohort of recently vaccinated individuals, subjects demonstrated robust total antibody and CD4+/CD8+ T cell response to SARS-CoV-2 mRNA vaccines when followed for 2 months post-2nd dose. In most individuals, T cell response declined between the 1st and 2nd doses suggesting a need for a booster or the completion of the 2-dose vaccine series. In a group of convalescent donors tested with QFN SARS-CoV-2 and Anti-CoV-2 tests, all patients had an antibody and T cell response up to 1 year after natural infection. CONCLUSION: This small feasibility study demonstrates that the QFN-SARS-CoV-2 test is able to identify CD4+ and CD8+ T cell-mediated responses in SARS-CoV-2-vaccinated subjects and those recovered from COVID-19, alongside a qualitative antibody response detectable via the QIAreach Anti-CoV2 test.

First performance report of QIAreach™ Anti-SARS-CoV-2 Total Test, an innovative nanoparticle fluorescence digital detection platform.

In 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused a global pandemic. Disease diagnosis, appropriate clinical management and infection control are all important factors in controlling the spread of SARS-CoV-2. The QIAreach™ Anti-SARS-CoV-2 Total Test (Anti-CoV2) is a rapid, qualitative serological test, using proprietary nanoparticle fluorescence technology to detect total antibody (IgA, IgM, and IgG) against SARS-CoV-2. Here we report the results of the US Food and Drug Administration (FDA) clinical agreement study. Thirty positive plasma or serum samples were taken from consenting individuals with polymerase chain reaction (PCR)-confirmed SARS-CoV-2 infection ≥14 days from symptom onset. Seventy-five samples from before the believed circulation of SARS-CoV-2 (November 1, 2019) were used to assess specificity. Positive percent agreement (PPA) and negative percent agreement (NPA) were calculated along with the corresponding exact two-sided 95 % confidence intervals (CI) using an FDA Emergency Use Authorized PCR test as the reference method. Anti-CoV2 was shown to have 100 % sensitivity (PPA; 95 % CI 88.4-100 %) and 100 % specificity (NPA; 95 % CI 95.2-100 %). Against 157 pre-pandemic samples, no cross-reactivity was observed with seasonal coronaviruses or other respiratory pathogens tested. Additionally, no interference was observed when samples were spiked with: conjugated bilirubin 0.4 mg/ml; unconjugated bilirubin 0.4 mg/ml; hemoglobin 5 mg/ml; prednisolone 0.12 mg/ml; triglycerides 15 mg/ml. In conclusion, Anti-CoV2 provides accurate qualitative detection of total antibodies against SARS-CoV-2.

Characterization of specific CD4 and CD8 T-cell responses in QuantiFERON TB Gold-Plus TB1 and TB2 tubes.

The new generation of the IGRA QuantiFERON-TB Gold Plus (QFT-Plus) includes two antigen tubes, TB1 and TB2 which contain specific Mycobacterium tuberculosis peptides designed to stimulate both CD4 and CD8 T-cells. TB1 is designed to target cell mediated responses from CD4 T-cells, while TB2 contains newly designed peptides designed to also stimulate CD8 T-cells. We identified specific CD4 and CD8 T-cell clones that recognize different regions spanning the length of the CFP-10 protein in M. tuberculosisto directly test in QFT-Plus TB1 and TB2 tubes, followed by Interferon-gamma detection by the QFT-Plus ELISA. These clones showed specific responses to the different QFT-Plus tubes, the CD4 T-cell clone showed dose-dependent responses to both TB1 and TB2 tubes, while the CD8 T-cell clones showed specific and targeted responses to the QFT-Plus TB2 tube (>140-fold difference) versus the QFT-Plus TB1 tubes using the QFT-Plus ELISA. This testing provides direct evidence of the specificity of CD8 T-cell mediated response in QFT-Plus TB2 tubes.

Psychometric testing of a person-centred care scale the Eden Warmth Survey in a long-term care home in New Zealand.

BACKGROUND: Traditional nursing homes have been viewed as dominated by the medical model. Since the 1990s, the Eden Alternative(TM) has become a significant model in systemic transformations in nursing homes. The purpose of this study was to evaluate the psychometric performance of the 20 items of the Eden Warmth Survey - Residents (EWS-R) in an aged-care home. DESIGN: A resident's satisfaction survey was used to collect a sample of 85 long-term care home residents. METHODS: Psychometric evaluation included item analyses, reliability including internal consistency and stability, criterion-related validity and construct validity. RESULTS: The reduced 13 items demonstrated adequate reliability (α = 0.82) with two factors, Trust and Connectedness with Others and Care Practices, extracted and contributed to 57.9% of the total variance. CONCLUSIONS: The 13-item of EWS-R can be considered as a reliable and predictive scale for assessing quality of life and overall satisfaction on people living in long-term care facilities.

Mechanisms and significance of reduced activity and responsiveness in resting frog tadpoles.

Hatchling Xenopus laevis tadpoles spend most of their time attached to objects or the water surface by mucus secreted by a gland on the head. While attached, swimming activity and responsiveness to swim-initiating stimuli are reduced over long periods of time. We have investigated the mechanisms and significance of this apparent long-term inhibition. In behavioural experiments we show, firstly, that innervation of the cement gland and GABA(A)-mediated inhibition are necessary for attachment to reduce responsiveness, and secondly, that denervation of the cement gland increases tadpole activity and increases their predation by damselfly nymphs (Zygoptera). To investigate the neuronal pathway from the cement gland to GABA(A) inhibition, we have devised an immobilized, inverted tadpole preparation where a weight attached to the mucus simulates the force as it hangs. Simulated attachment reduces responsiveness and spontaneous fictive swimming activity. We have recorded the activity and responses of trigeminal neurons innervating the cement gland. They are spontaneously active and simulating attachment results in a sustained increase in this activity. We propose that hanging from a mucus strand increases firing in cement gland afferents. This leads to tonic GABA inhibition that reduces tadpole activity and responses, and leads to fewer attacks by predators.

Validating notifications of pregnancy terminations for birth defects before 20 weeks gestation.

The Victorian Perinatal Data Collection Unit (PDCU) maintains a Birth Defects Register (BDR), including data on terminations of pregnancy for birth defects before 20 weeks. This study aimed to validate the completeness of reporting, and accuracy of diagnoses provided by hospitals on these terminations. From 1995-1999, 13 hospitals notified 321 terminations to the PDCU. An additional 41 cases were found through this study. For matched cases (n=273), 199 (73%) had comparable diagnoses, 21 (8%) had different and 53 (19%) had additional diagnoses. Discrepancies arose because of problems with correct identification of relevant cases and lack of understanding of the importance of including all information on multiple birth defects.