Early Adoption of Longitudinal Surveillance for SARS-CoV-2 among Staff in Long-Term Care Facilities: Prevalence, Virologic and Sequence Analysis

Gallichotte, Emily N.; Quicke, Kendra M.; Sexton, Nicole R.; Fitzmeyer, Emily; Young, Michael C.; Janich, Ashley J; Dobos, Karen M.; Pabilonia, Kristy L.; Gahm, Gregory A.; Carlton, Elizabeth J.; Ebel, Gregory D.; Ehrhart, Nicole

doi:10.1128/spectrum.01003-21

Cited by 25 publications

(38 citation statements)

References 48 publications

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“…qrt-PCR was performed using the EXPRESS One-Step SuperScript qrt-PCR reagents and N1, N2, E, and human RNAse P primer/probes as previously described. 13 …”

Section: Methodsmentioning

confidence: 99%

“…Sequencing and analyses were performed as previously described. 13 Briefly, cDNA was generated, PCR amplification was performed using ARTIC primers and Q5 High-Fidelity polymerase, products were purified and libraries were prepared using KAPA HyperPrep kit and unique index primers. Libraries were sequenced on the Illumina MiSeq V2 using 2 × 250 paired-end reads.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Detection of SARS-CoV-2 in exhaled air using non-invasive embedded strips in masks

Gallichotte¹,

Windsor²,

Watts³

et al. 2022

American Journal of Infection Control

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“…qrt-PCR was performed using the EXPRESS One-Step SuperScript qrt-PCR reagents and N1, N2, E, and human RNAse P primer/probes as previously described. 13 …”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Detection of SARS-CoV-2 in exhaled air using non-invasive embedded strips in masks

Gallichotte¹,

Windsor²,

Watts³

et al. 2022

American Journal of Infection Control

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“…Surveillance testing was performed as previously described [8, 9]. Briefly, nasal swabs were collected, processed, viral RNA extracted, and quantitative reverse transcriptase PCR (qPCR) was performed using the Thermo Fisher Scientific TaqPath COVID-10 combo kit, under U.S. FDA Emergency Use Authorization [17].…”

Section: Methodsmentioning

confidence: 99%

“…Due to the high number of cases in LTCFs prior to vaccines and other preventative measures, many staff and residents became infected during 2020 and 2021, with some facilities reporting infection and seroprevalence rates as high as 40% [8][9][10][11]. Therefore, there are two immunologically distinct populations of individuals receiving vaccines; those that are naïve, with no evidence of a prior infection (seronegative), and those that are pre-immune with either a documented prior infection, or serological evidence of prior infection (seropositive).…”

Section: Introductionmentioning

confidence: 99%

Impact of prior infection on SARS-CoV-2 antibody responses in vaccinated long-term care facility staff

Gallichotte

Nehring

Stromberg

et al. 2022

Preprint

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SARS-CoV-2 emerged in 2019 and has resulted in millions of deaths worldwide. Certain populations are at higher risk for infection, especially staff and residents at long term care facilities (LTCF), due to the congregant living setting, and residents with many comorbidities. Prior to vaccine availability, these populations represented a large fraction of total COVID-19 cases and deaths in the U.S. Due to the high-risk setting and outbreak potential, staff and residents were among the first groups to be vaccinated. To define the impact of prior infection on response to vaccination, we measured antibody responses in a cohort of staff members at a LTCF, many of whom were previously infected by SARS-CoV-2. We found that neutralizing, receptor-binding-domain (RBD) and nucleoprotein (NP) binding antibody levels were significantly higher post-full vaccination course in individuals that were previously infected, and NP antibody levels could discriminate individuals with prior infection from vaccinated individuals. While an anticipated antibody titer increase was observed after vaccine booster dose in naïve individuals, boost response was not observed in individuals with previous COVID-19 infection. We observed a strong relationship between neutralizing antibodies and RBD-binding antibodies post-vaccination across all groups, suggesting RBD-binding antibodies may be used as a correlate of neutralization. One individual with high levels of neutralizing and binding antibodies experienced a breakthrough infection (prior to the introduction of Omicron), demonstrating that the presence of antibodies is not always sufficient for complete protection against infection. These results highlight that history of COVID-19 exposure significantly increases SARS-CoV-2 antibody responses following vaccination.ImportanceLong-term care facilities (LTCFs) have been disproportionately impacted by COVID-19, due to their communal nature, high-risk profile of residents and vulnerability to respiratory pathogens. In this study, we analyzed the role of prior natural immunity to SARS-CoV-2 on post-vaccination antibody responses. The LTCF in our cohort experienced a large outbreak with almost 40% of staff becoming infected. We found that individuals that were infected prior to vaccination, had higher levels of neutralizing and binding antibodies post-vaccination. Importantly, the second vaccine dose significantly boosted antibody levels in those that were immunologically naïve prior to vaccination, but not those that had prior immunity. Regardless of pre-vaccination immune status, levels of binding and neutralizing antibodies were highly correlated. The presence of NP-binding antibodies can be used to identify individuals that were previously infected when pre-vaccination immune status is not known. Our results reveal that vaccination antibody responses differ depending on prior natural immunity.

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“…Nasopharyngeal swabs were collected from asymptomatic staff at long-term care facilities and characterized as described here. 37 Briefly, viral RNA was extracted and quantified using qPCR with CDC primer probes. The infectious virus was measured using a standard plaque assay on Vero cells starting with 250 μL of input material.…”

Section: Experimental Sectionmentioning

confidence: 99%

Electrochemical Immunoassay for the Detection of SARS-CoV-2 Nucleocapsid Protein in Nasopharyngeal Samples

et al. 2022

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Point-of-care (POC) methods currently available for detecting SARS-CoV-2 infections still lack accuracy. Here, we report the development of a highly sensitive electrochemical immunoassay capable of quantitatively detecting the presence of the SARS-CoV-2 virus in patient nasopharyngeal samples using stencil-printed carbon electrodes (SPCEs) functionalized with capture antibodies targeting the SARS-CoV-2 nucleocapsid protein (N protein). Samples are added to the electrode surface, followed by horseradish peroxidase (HRP)-conjugated detection antibodies also targeting the SARS-CoV-2 N protein. The concentration of the virus in samples is quantified using chronoamperometry in the presence of 3,3′5,5′-tetramethylbenzidine. Limits of detection equivalent to less than 50 plaque forming units/mL (PFU/mL) were determined with virus sample volumes of 20 μL. No cross-reactivity was detected with the influenza virus and other coronavirus N proteins. Patient nasopharyngeal samples were tested as part of a proof-of-concept clinical study where samples were also tested using the gold-standard real-time quantitative polymerase chain reaction (RT-qPCR) method. Preliminary results from a data set of 22 samples demonstrated a clinical specificity of 100% ( n = 9 negative samples according to RT-qPCR) and a clinical sensitivity of 70% for samples with RT-PCR cycle threshold (Ct) values under 30 ( n = 10) and 100% for samples with Ct values under 25 ( n = 5), which complies with the World Health Organization (WHO) criteria for POC COVID-19 diagnostic tests. Our functionalized SPCEs were also validated against standard plaque assays, and very good agreement was found between both methods ( R 2 = 0.9993, n = 6), suggesting that our assay could be used to assess patient infectivity. The assay currently takes 70 min from sampling to results.

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Early Adoption of Longitudinal Surveillance for SARS-CoV-2 among Staff in Long-Term Care Facilities: Prevalence, Virologic and Sequence Analysis

Abstract: Our work comprises unique data on the characteristics of SARS-CoV-2 dynamics among staff working at LTCFs in the early months of the SARS-CoV-2 pandemic prior to mandated staff surveillance testing. During this time period, LTCF residents were largely sheltering-in-place.

Cited by 25 publications

References 48 publications

Detection of SARS-CoV-2 in exhaled air using non-invasive embedded strips in masks

Detection of SARS-CoV-2 in exhaled air using non-invasive embedded strips in masks

Impact of prior infection on SARS-CoV-2 antibody responses in vaccinated long-term care facility staff

Electrochemical Immunoassay for the Detection of SARS-CoV-2 Nucleocapsid Protein in Nasopharyngeal Samples

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