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Diagnosis of ischemic stroke using circulating levels of brain-specific proteins measured via high-sensitivity digital ELISA.

ABSTRACT: Limited lower detection ranges associated with traditional immunoassay techniques have prevented the use of brain-specific proteins as blood biomarkers of stroke in the acute phase of care, as these proteins are often only present in circulation at low concentrations. Digital ELISA is a newly developed technique with allows for quantification of proteins in biofluids with up to 1000 times greater sensitivity than conventional ELISA techniques. The purpose of this study was to determine whether the extended lower limits of detection associated with digital ELISA could enable the use of brain-specific proteins as blood biomarkers of ischemic stroke during triage. Blood was sampled from ischemic stroke patients (n = 14) at emergency department admission, as well as from neurologically normal controls matched in terms of risk factors for cardiovascular disease (n = 33). Plasma levels of two brain-specific axonal proteins, neurofilament light chain (NfL) and tau, were measured via digital ELISA, and receiver-operating characteristic analysis was used to determine their ability to discriminate between groups. Plasma levels of NfL and tau were both significantly elevated in stroke patients versus controls, and could respectively discriminate between groups with 92.9% sensitivity / 84.9% specificity, and 85.7% sensitivity / 54.6% specificity. Furthermore, adjustment of measured NfL and Tau levels according to the lower-limits of detection associated with commercially-available conventional ELISA assays resulted in a dramatic and statistically significant decrease in diagnostic performance. Collectively, our results suggest that the increased analytical sensitivity of digital ELISA could enable the use of brain-specific proteins as blood biomarkers of ischemic stroke during triage.

SUBMITTER: O'Connell GC

PROVIDER: S-EPMC7409378 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The precision by which animal diseases are diagnosed affects our ability to make informed decisions with regards to animal health management, from a clinical and economic perspective. Lameness is a major health condition in dairy cattle. The underlying causes of lameness include bovine digital dermatitis (BDD), which is reported as one of the main causes of infectious lameness in dairy cattle. Presently, the gold standard for BDD diagnosis in dairy cattle is visual inspection of lifted hooves-a labour intensive and subjective method. Research has suggested that Treponema spp. are the main pathogens associated with the establishment of BDD. We explored the potential of indirect enzyme-linked immunosorbent assay (ELISA) as a diagnostic serological tool in the identification of cows at different stages of BDD. Additionally, we evaluated the predictive power of this diagnostic tool on the future occurrence of BDD lesions. A total of 232 cows from three farms were used in the study. Serum samples and hoof health data were collected at three time points: ~ 30 days pre-calving, around calving, and approximately 30 days post-calving. The mean absorbance from the ELISA test was compared across different clinical presentations of BDD as assessed by visual inspection of the hooves according to the M-stage classification system. A transition model was developed to estimate the probability of lesion occurrence in time t + 1 based on the spectrophotometer (absorbance) reading in time t. The mean absorbance reading for both IgG1 and IgG2 anti-Treponema antibodies was associated with disease presence-apart from M4.1 lesions, animals with no lesions had a lower mean when compared to animals with lesions regardless of the score. Additionally, the mean absorbance reading of animals with active lesions was higher when compared to animals with no lesions. However, the anti-Treponema antibody assays failed to identify disease presence in a consistent manner. Moreover, indirect ELISA readings were not a predictor of the future occurrence of BDD lesions. In conclusion, although the levels anti-Treponema antibodies were associated with disease presence, the ELISA test failed to detect disease unequivocally and had no predictive value in the future occurrence of BDD lesions.

Project description:BackgroundTo determine whether dizziness can contribute to stroke as a main cause still remains challenging. This study aims to explore clinical biomarkers in the identification of ischemic stroke patients from people with dizziness and the prediction of their long-term recovery.MethodsFrom January 2018 to June 2019, 21 ischemic stroke patients with a main complaint of dizziness, 84 non-stroke dizziness patients and 87 healthy volunteers were recruited in this study. Then, their peripheral blood samples were collected, and the percentages of circulating lymphocytes T cells, CD4+ T cells, CD8+ T cells, T-/- cells (DNTs), CD4+ regulatory T cells (Tregs), CD8+ Tregs, B cells and regulatory B cells (Bregs) were examined to identify biomarkers with clinical value.ResultsAccording to our data, a significant difference in the DNTs proportion was detected between non-stroke dizziness and ischemic stroke patients with dizziness (p = 0.0009). The Bregs proportion in ischemic stroke patients with dizziness was lower than that in non-stroke dizziness patients (p = 0.035). In addition, the percentage of Bregs and DNTs within lymphocytes in patients' peripheral blood exhibited a significant negative correlation with stroke occurrence (Bregs, p = 0.039; DNTs, p = 0.046). Moreover, the Bregs and DNTs within lymphocytes were negatively related to participants' age, while presented a weak relationship with clinical risks like smoking, hypertension, and diabetes. Then, area under the receiver operating characteristic curve (AUC) of Bregs and DNTs together was 0.768, the risk factors and Bregs or DNTs ranged from 0.795 and 0.792, respectively, and the AUC value of risk factors, Bregs and DNTs combination was further increased to 0.815. Furthermore, the Bregs percentage within lymphocytes at admission was also a potential predictor of repair at discharge and the following 3 months.ConclusionBregs and DNTs could be the clinical biomarkers together in the identification of ischemic stroke patients from people with dizziness.

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Diagnosis of ischemic stroke using circulating levels of brain-specific proteins measured via high-sensitivity digital ELISA.

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