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  • 1
    Online Resource
    Online Resource
    Ultrasensitive assay for saliva-based SARS-CoV-2 antigen detection
    Walter de Gruyter GmbH ; 2022
    In:  Clinical Chemistry and Laboratory Medicine (CCLM) Vol. 60, No. 5 ( 2022-04-26), p. 771-777
    Details
    In: Clinical Chemistry and Laboratory Medicine (CCLM), Walter de Gruyter GmbH, Vol. 60, No. 5 ( 2022-04-26), p. 771-777
    Abstract: Widespread SARS-CoV-2 testing is invaluable for identifying asymptomatic/pre-symptomatic individuals. There remains a technological gap for highly reliable, easy, and quick SARS-CoV-2 diagnostic tests suitable for frequent mass testing. Compared to nasopharyngeal (NP) swab-based tests, saliva-based methods are attractive due to easier and safer sampling. Current saliva-based SARS-CoV-2 rapid antigen tests (RATs) are hindered by limited analytical sensitivity. Here, we report one of the first ultrasensitive, saliva-based SARS-CoV-2 antigen assays with an analytical sensitivity of 〈 0.32 pg/mL, corresponding to four viral RNA copies/µL, which is comparable to that of PCR-based tests. Methods Using the novel electrochemiluminescence (ECL)-based immunoassay, we measured the SARS-CoV-2 nucleocapsid (N) antigen concentration in 105 salivas, obtained from non-COVID-19 and COVID-19 patients. We then verified the results with a second, independent cohort of 689 patients (3.8% SARS-CoV-2 positivity rate). We also compared our method with a widely used point-of-care rapid test. Results In the first cohort, at 100% specificity, the sensitivity was 92%. Our assay correctly identified samples with viral loads up to 35 CT cycles by saliva-based PCR. Paired NP swab-based PCR results were obtained for 86 cases. Our assay showed high concordance with saliva-based and NP swab-based PCR in samples with negative ( 〈 0.32 pg/mL) and strongly positive ( 〉 2 pg/mL) N antigen concentrations. In the second cohort, at 100% specificity, sensitivity was also 92%. Our assay is about 700-fold more sensitive than the Abbott Panbio Rapid Test. Conclusions We demonstrated the ultrasensitivity and specificity assay and its concordance with PCR. This novel assay is especially valuable when compliance to frequent swabbing may be problematic.
    Type of Medium: Online Resource
    ISSN: 1434-6621 , 1437-4331
    URL: Article
    DOI: 10.1515/cclm-2021-1142
    Language: English
    Publisher: Walter de Gruyter GmbH
    Publication Date: 2022
    detail.hit.zdb_id: 1492732-9
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  • 2
    Online Resource
    Online Resource
    Quantitation of neurofilament light chain protein in serum and cerebrospinal fluid from patients with multiple sclerosis using the MSD R-PLEX NfL assay
    Walter de Gruyter GmbH ; 2023
    In:  Diagnosis Vol. 10, No. 3 ( 2023-08-14), p. 275-280
    Details
    In: Diagnosis, Walter de Gruyter GmbH, Vol. 10, No. 3 ( 2023-08-14), p. 275-280
    Abstract: Neurofilament light (NfL) chain is a marker of neuroaxonal damage in various neurological diseases. Here we quantitated NfL levels in the cerebrospinal fluid (CSF) and serum from patients with multiple sclerosis (MS) and controls, using the R-PLEX NfL assay, which employs advanced Meso Scale Discovery ® (MSD) electrochemiluminescence (ECL)-based detection technology. Methods NfL was quantitated in samples from 116 individuals from two sites (Ottawa Hospital Research Institute and Mayo Clinic), consisting of patients with MS (n=71) and age- and sex-matched inflammatory neurological controls (n=13) and non-inflammatory controls (n=32). Correlation of NfL levels between CSF and serum was assessed in paired samples in a subset of MS patients and controls (n=61). Additionally, we assessed the correlation between NfL levels obtained with MSD’s R-PLEX ® and Quanterix’s single molecule array (Simoa ® ) assays in CSF and serum (n=32). Results Using the R-PLEX, NfL was quantitated in 99% of the samples tested, and showed a broad range in the CSF (82–500,000 ng/L) and serum (8.84–2,014 ng/L). Nf-L levels in both biofluids correlated strongly (r=0.81, p 〈 0.0001). Lastly, Nf-L measured by MSD’s R-PLEX and Quanterix’s Simoa assays were highly correlated for both biofluids (CSF: r=0.94, p 〈 0.0001; serum: r=0.95, p 〈 0.0001). Conclusions We show that MSD’s R-PLEX NfL assay can reliably quantitate levels of NfL in the CSF and serum from patients with MS and controls, where levels correlate strongly with Simoa.
    Type of Medium: Online Resource
    ISSN: 2194-802X
    URL: Article
    DOI: 10.1515/dx-2022-0125
    Language: English
    Publisher: Walter de Gruyter GmbH
    Publication Date: 2023
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  • 3
    Online Resource
    Online Resource
    Sensitive Serology Measurements in the Saliva of Individuals with COVID-19 Symptoms Using a Multiplexed Immunoassay
    Oxford University Press (OUP) ; 2022
    In:  The Journal of Applied Laboratory Medicine Vol. 7, No. 6 ( 2022-10-29), p. 1354-1365
    Details
    In: The Journal of Applied Laboratory Medicine, Oxford University Press (OUP), Vol. 7, No. 6 ( 2022-10-29), p. 1354-1365
    Abstract: There are numerous benefits to performing salivary serology measurements for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative pathogen for coronavirus disease 2019 (COVID-19). Here, we used a sensitive multiplex serology assay to quantitate salivary IgG against 4 SARS-CoV-2 antigens: nucleocapsid, receptor-binding domain, spike, and N-terminal domain. Methods We used single samples from 90 individuals with COVID-19 diagnosis collected at 0 to 42 days postsymptom onset (PSO) and from 15 uninfected control subjects. The infected individuals were segmented in 4 groups (0–7 days, 8–14 days, 15–21 days, and & gt;21 days) based on days PSO, and values were compared to controls. Results Compared to controls, infected individuals showed higher levels of antibodies against all antigens starting from 8 days PSO. When applying cut-offs with at least 93.3% specificity at every time interval segment, nucleocapsid protein serology had the best sensitivity at 0 to 7 days PSO (60% sensitivity [35.75% to 80.18%], ROC area under the curve [AUC] = 0.73, P = 0.034). Receptor-binding domain serology had the best sensitivity at 8 to 14 days PSO (83.33% sensitivity [66.44%–92.66%], ROC AUC = 0.90, P & lt; 0.0001), and all assays except for N-terminal domain had 92% sensitivity (75.03%–98.58%) at & gt;14 days PSO. Conclusions This study shows that our multiplexed immunoassay can distinguish infected from uninfected individuals and reliably (93.3% specificity) detect seroconversion (in 60% of infected individuals) as early as the first week PSO, using easy-to-collect saliva samples.
    Type of Medium: Online Resource
    ISSN: 2475-7241
    URL: Article
    DOI: 10.1093/jalm/jfac073
    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2022
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