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Low-volume antibody assays can be used to track SARS-CoV-2 infection rates in settings where active testing for virus is limited and remote sampling is optimal. We developed 12 ELISAs detecting total or antibody isotypes to SARS-CoV-2 nucleocapsid, spike protein or its receptor binding domain (RBD), 3 anti-RBD isotype specific luciferase immunoprecipitation system (LIPS) assays and a novel Spike-RBD bridging LIPS total-antibody assay. We utilized pre-pandemic (n=984) and confirmed/suspected recent COVID-19 sera taken pre-vaccination rollout in 2020 (n=269). Assays measuring total antibody discriminated best between pre-pandemic and COVID-19 sera and were selected for diagnostic evaluation. In the blind evaluation, two of these assays (Spike Pan ELISA and Spike-RBD Bridging LIPS assay) demonstrated >97% specificity and >92% sensitivity for samples from COVID-19 patients taken >21 days post symptom onset or PCR test. These assays offered better sensitivity for the detection of COVID-19 cases than a commercial assay which requires 100-fold larger serum volumes. This study demonstrates that low-volume in-house antibody assays can provide good diagnostic performance, and highlights the importance of using well-characterized samples and controls for all stages of assay development and evaluation. These cost-effective assays may be particularly useful for seroprevalence studies in low and middle-income countries.
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http://dx.doi.org/10.3389/fimmu.2022.968317 | DOI Listing |
J Am Soc Nephrol
September 2025
Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA.
Background: Genetic modifiers are believed to play an important role in the onset and severity of polycystic kidney disease (PKD), but identifying these modifiers has been challenging due to the lack of effective methodologies.
Methods: We generated zebrafish mutants of IFT140, a skeletal ciliopathy gene and newly identified autosomal dominant PKD (ADPKD) gene, to examine skeletal development and kidney cyst formation in larval and juvenile mutants. Additionally, we utilized ift140 crispants, generated through efficient microhomology-mediated end joining (MMEJ)-based genome editing, to compare phenotypes with mutants and conduct a pilot genetic modifier screen.
Med Vet Entomol
September 2025
Entomology Research Unit, Department of Zoology, The University of Burdwan, Burdwan, India.
The biting midges, Culicoides peregrinus Kieffer and Culicoides oxystoma Kieffer (Diptera: Ceratopogonidae) are the most significant vector species of bluetongue virus (BTV) in the Oriental region, including India. Rearing of these vector species was cumbersome; previous researchers supplemented the rearing substrates primarily with cattle dung (the habitat), yeast and nutrient broth. Other investigations reiterated that an enriched milieu of live bacteria is required for the oviposition and developmental progression of the immatures as they failed to develop in sterile medium.
View Article and Find Full Text PDFProc Natl Acad Sci U S A
September 2025
State Key Laboratory of Bioactive Molecules and Druggability Assessment, Guangdong Province Key Laboratory of Pharmacodynamic Constituents of Traditional Chinese Medicine and New Drugs Research, International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug De
Proliferative retinopathy is a leading cause of irreversible blindness in humans; however, the molecular mechanisms behind the immune cell-mediated retinal angiogenesis remain poorly elucidated. Here, using single-cell RNA sequencing in an oxygen-induced retinopathy (OIR) model, we identified an enrichment of sorting nexin (SNX)-related pathways, with SNX3, a member of the SNX family that is involved in endosomal sorting and trafficking, being significantly upregulated in the myeloid cell subpopulations of OIR retinas. Immunostaining showed that SNX3 expression is markedly increased in the retinal microglia/macrophages of mice with OIR, which is mainly located within and around the neovascular tufts.
View Article and Find Full Text PDFJAMA Netw Open
September 2025
Perinatal Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
Importance: Exposure to inflammation from chorioamnionitis places the fetus at higher risk of premature birth and may increase the risk of neurodevelopmental impairments, though the evidence for the latter is mixed.
Objective: To evaluate whether moderate to severe histologic chorioamnionitis (HCA) is directly associated with adverse motor performance, independent of the indirect mediating effects of premature birth.
Design, Setting, And Participants: This prospective, population-based cohort study recruited participants between September 16, 2016, and November 19, 2019, from referral and nonreferral neonatal intensive care units of 5 southwestern Ohio hospitals.