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Objetivo: Evaluar la validez diagnóstica del ensayo de inmunoabsorción ligado a enzima (Enzyme-Linked Immunosorbent Assay, elisa) para el virus de inmunodeficiencia humana (vih) en bancos de sangre, con base en estudios publicados entre 2000 y 2020. Metodología: Se realizó una revisión sistemática y metaanálisis de pruebas diagnósticas, mediante un modelo de efectos aleatorios para la sensibilidad, la especificidad, el cociente de probabilidad positivo y negativo, la razón de odds (or) diagnóstica y la curva roc, con sus intervalos de confianza del 95 %. La heterogeneidad se evaluó con el estadístico Q(χ2) DerSimonian-Laird y el I2 de inconsistencia, y la incertidumbre, con el porcentaje de peso de cada estudio. Resultados: Se incluyeron 15 investigaciones; la elisa de tercera generación (detección de anticuerpos) se aplicó en 2992 infectados y 4076 sanos; las de cuarta generación (determinación simultánea de antígeno-anticuerpo), en 967 infectados y 154 264 sanos; ambas presentaron sensibilidad cercana al 100 %, pero la especificidad fue mejor en los ensayos de cuarta generación (98 vs. 100 %). Para ambas tecnologías, los cocientes de probabilidad, or diagnóstica y curva roc evidenciaron excelente discriminación de sanos e infectados. Conclusión: Se confirmó que las elisa de tercera y cuarta generación presentan excelente validez y utilidad diagnóstica en donantes de sangre, lo que es importante para las políticas de sangre segura y control del vih.

Bioimprinting was performed against ovalbumin (OVA) to confer its binding cavities for kwakhurin (Kwa), an isoflavonoid, produced solely by Pueraria candollei var. mirifica (P. candollei). The characterization of bioimprinted-OVA (biOVA), evaluated by an enzyme-linked immunosorbent assay (ELISA), revealed that it functioned as a specific receptor for Kwa. Using biOVA, two systems, i.e., an indirect competitive ELISA (icELISA) and the even simpler and more rapid competitive enzyme-linked bioimprinted-protein assay (cELBIA), were developed as novel techniques for the quantitative analysis of Kwa in P. candollei and its related products. The two analysis methods were found to have limits of detection (LOD) of 4.0 and 2.5 µg/mL, respectively. The high reliability of the developed icELISA and cELBIA using biOVA was also demonstrated by various validation analyses. Subsequently, bioimprinting was performed using eight other proteins to investigate them as candidate scaffolds for the generation of binding cavities for Kwa. Interestingly, two bioimprinted-IgG monoclonal antibodies (biMAbs) recognized Kwa, but their original binding affinity to hapten was lost. That is, the MAbs obtained a new binding ability to Kwa in exchange for their original binding affinity, raising the possibility that biMAb could be alternatively used as a probe for the quantitative analysis of Kwa as well as biOVA. This is the first report of small molecules recognition by MAbs used as proteins for bioimprinting.

Background The programmed cell death‐1/programmed cell death‐1 ligand (PD‐1/PD‐L1) pathway plays a crucial role in tumor evasion. This study evaluated the association between circulating PD‐L1 expression and clinical characteristics in patients with advanced non‐small cell lung cancer (NSCLC). Methods A total of 109 advanced NSCLC and 65 healthy patients from the Beijng Cancer Hospital were enrolled in the study. Circulating PD‐L1 expression was tested by enzyme‐linked immunosorbent assay. The associations between the level of PD‐L1 expression and clinicopathologic features and prognosis were statistically analyzed. Results The expression of PD‐L1 in advanced NSCLC patients was significantly upregulated compared with the healthy control (P < 0.001). The expression of PD‐L1 was significantly correlated with abdominal organ metastasis (P = 0.004). A high PD‐L1 expression had a worse prognosis than a low expression in patients (18.7 vs. 26.8 month, P < 0.001). Conclusions PD‐L1 was elevated in advanced NSCLC patients and may play an important role in tumor immune evasion and patient prognosis.

ABSTRACT Cavin‐4 was identified as a potential autoantigen for immune‐mediated rippling muscle disease (iRMD). To validate this, we developed and tested various immunoassays, including a cell‐based assay (CBA), cavin‐4 recombinant protein ELISA, and multi‐peptide ELISA. Among 19 iRMD patients, all exhibited muscle rippling, and 13 had percussion‐induced mounding. All immunoassays demonstrated clinical and analytical specificities greater than 95%. The protein ELISA had the highest sensitivity (94.7%) and specificity (99.9%), outperforming CBA (sensitivity 89.5%, specificity 99.6%) and the multi‐peptide ELISA (sensitivity 79.0%, specificity 97.2%). Our results suggest that the cavin‐4 protein ELISA is a promising tool for high‐throughput clinical testing in iRMD.

ABSTRACT Purpose Infectious bovine rhinotracheitis (IBR) is one of the most important diseases affecting production and productivity. Methodology Cross‐sectional study was aimed at to determine the seroprevalence of IBR and associated risk factors, and animal owners’ knowledge, attitude and practice towards the disease from April 2021 to June 2022. Accordingly, a total of 384 serum samples were collected from both crossbreed (70) and local breed (314) cattle from purposively selected districts of East Wollega zone of Western Ethiopia. Competitive enzyme‐linked immunosorbent assay (ELISA) was used for testing glycoprotein antibodies (anti‐gB) for bovine herpes virus‐1 (BoHV‐1) virus in collected serum, and the obtained data were analysed by multiple logistic regressions by using R software 3.62 version. However, questionnaire data were analysed for descriptive statistics by SPSS version 20.0 (IBM. Corp, 2011). Result The total prevalence of IBR in the study area was found to be 70.54% at herd and 80.47% at individual cattle level. The significant association (p <0.05$ < \\ 0.05$) was found for breed, age, body condition and herd size but not for district and sex as risk factors. The BoHV‐1 virus seropositivity in adult animals increased significantly, with an odds ratio of 1.65 (95% CI 0.705–3.85) compared to young. Local breed cattle were 2.055 times more likely to test positive for IBR with an odds ratio of 0.77 (95% CI 0.23–2.22) compared to crossbreed cattle. The chances of cattle in medium herds testing positive for the BoHV‐1 virus with an odds ratio of (1.78 95% CI 1.303–7.50) are greater than the chances of cattle in smaller herds testing positive. The survey results showed that 70% of animal owners identified IBR as a major challenge in animal production, whereas 35% mentioned long calving intervals. However, 92% of the participants were not informed about the level of knowledge and attitude regarding particular diseases such as IBR. Conclusion This study showed that there is a high prevalence of IBR in cattle in the study area, and that owners have low awareness of the disease. Therefore, it is necessary to develop an immediate control system and conduct additional research on molecular detection to evaluate its effects on reproductive performance. Cross‐sectional study was used to determine the seroprevalence of IBR, associated risk factors and KAP. A total of 384 cattle's sera samples were collected and competitive ELISA for testing anti‐gB. A total of 100 questionnaire data were collected from animal owners. The total prevalence was 70.54% at herd and 80.47% at individual cattle level. It was influenced by age, body condition, breed and location. Overall, 92% of animal owners had no awareness towards specific diseases like IBR. Hence, control mechanism and further studies should be designed.

Human performance enhancing drugs (PEDs), frequently used in sport competitions, are strictly prohibited by the World Anti-Doping Agency (WADA). Biological samples collected from athletes and regular patients are continuously tested regarding the identification and/or quantification of the banned substances. Current work is focused on the application of a new analytical method, molecularly imprinted nanoparticles (nanoMIPs), to detect and determine concentrations of certain prohibited drugs, such as β-blockers, in water and human urine samples. These medications are used in the treatment of cardiovascular conditions, negative effects of adrenaline (helping to relief stress), and hypertension (slowing down the pulse and softening the arteries). They can also significantly increase muscle relaxation and improve heart efficiency. The new method of the detection and quantification of β-blockers is based on synthesis, characterization, and implementation of nanoMIPs (so-called plastic antibodies). It offers numerous advantages over the traditional methods, including high binding capacity, affinity, and selectivity for target molecules. Additionally, the whole process is less complicated, cheaper, and better controlled. The size and shape of the nanoMIPs is evaluated by dynamic light scattering (DLS) and transmission electron microscope (TEM). The affinity and selectivity of the nanoparticles are investigated by competitive pseudo enzyme-linked immunosorbent assay (pseudo-ELISA) similar to common immunoassays employing natural antibodies. To provide reliable results towards either doping detection or therapeutic monitoring using the minimal invasive method, the qualitative and quantitative analysis of these drugs is performed in water and human urine samples. It is demonstrated that the assay can detect β-blockers in water within the linear range 1 nmol·L−1–1 mmol·L−1 for atenolol with the detection limit 50.6 ng mL−1, and the linear range 1 mmol·L−1–10 mmol·L−1 for labetalol with the detection limit of 90.5 ng·mL−1. In human urine samples, the linear range is recorded in the concentration range 0.1 mmol·L−1–10 nmol·L−1 for atenolol and 1 mmol·L−1–10 nmol·L−1 for labetalol with a detection limit of 61.0 ng·mL−1 for atenolol and 99.4 ng·mL−1 for labetalol.

In this study, an extremely highly sensitive enzyme-linked immunosorbent assay (ELISA) based on a newly produced monoclonal antibody (mAb) for the detection of ochratoxin A (OTA) in food samples was developed. OTA-Bovine serum albumin (BSA) conjugate was prepared and used as the immunogen for the production of the mAb. Among four hybridoma clones (8B10, 5C2, 9B7, and 5E11), the antibody from 8B10 displayed the highest affinity recognition for OTA. Based on the mAb (8B10), the IC[sub.50] and LOD of the ELISA for OTA were 34.8 pg mL[sup.−1] and 1.5 pg mL[sup.−1], respectively, which was 1.53~147 times lower than those in published ELISAs, indicating the ultra-sensitivity of our assay. There was no cross-reactivity of the mAb with the other four mycotoxins (AFB[sub.1], ZEN, DON, and T-2). Due to the high similarity in molecular structures among OTA, ochratoxin B (OTB), and ochratoxin C (OTC), the CR values of the mAb with OTB and OTC were 96.67% and 22.02%, respectively. Taking this advantage, the ELISA may be able to evaluate total ochratoxin levels in food samples. The recoveries of the ELISA for OTA in spiked samples (corn, wheat, and feed) were 96.5–110.8%, 89.5–94.4%, and 91.8–113.3%; and the RSDs were 5.2–13.6%, 8.2–13.0%, and 7.7–13.7% (n = 3), respectively. The spiked food samples (corn) were measured by ELISA and HPLC-FLD simultaneously. A good correlation between ELISA (x) and HPLC-FLD (y) with the linear regression equation of y = 0.918x − 0.034 (R[sup.2] = 0.985, n = 5) was obtained. These results demonstrated that the newly produced mAb-based ELISA was a feasible and ultra-sensitive analytical method for the detection of OTA in food samples.

Identification of smokers having predisposition to COPD is important for early intervention to reduce the huge global burden of the disease. Using a guinea pig model, we have shown that -benzoquinone ( -BQ) derived from cigarette smoke (CS) in the lung is a causative factor for CS-induced emphysema. -BQ is also derived from CS in smokers and it elicits the production of anti- -BQ antibody in humans. We therefore hypothesized that anti- -BQ antibody might have a protective role against COPD and could be used as a predictive biomarker for COPD in smokers. The objective of this study was to compare the serum anti- -BQ antibody level between smokers with and without COPD for the evaluation of the hypothesis. Serum anti- -BQ antibody concentrations of current male smokers with (n=227) or without (n=308) COPD were measured by an indirect enzyme-linked immunoabsorbent assay (ELISA) developed in our laboratory. COPD was diagnosed by spirometry according to Global Initiative for Chronic Obstructive Lung Disease (GOLD) guidelines. A significant difference was observed in the serum anti- -BQ antibody level between smokers with and without COPD (Mann-Whitney -test =4,632.5, =0.000). Receiver operating characteristic (ROC) curve analysis indicated that the ELISA had significant precision (area under the curve [AUC] =0.934, 95% confidence interval [CI]: 0.913-0.935) for identifying smokers with COPD from their low antibody level. The antibody cutoff value of 29.4 mg/dL was constructed from the ROC coordinates to estimate the risk for COPD in smokers. While 90.3% of smokers with COPD had a low antibody value (≤29.4 mg/dL), the majority (86.4%) of smokers without COPD had a high antibody value (≤29.4 mg/dL); 13.6% of current smokers without COPD having an antibody level below this cutoff value (odds ratio [OR] =59.3, 95% CI: 34.15-101.99) were considered to be at risk for COPD. Our results indicate that serum anti- -BQ antibody level may be used as a biomarker to identify asymptomatic smokers at risk for COPD for early intervention of the disease.