Hepatocellular carcinoma diagnosis using a novel electrochemiluminescence immunoassay targeting serum IgM-free AIM

Background Recent increases in the number of patients with non-alcoholic steatohepatitis (NASH) warrant the identification of biomarkers for early detection of hepatocellular carcinoma (HCC) associated with NASH (NASH-HCC). IgM-free apoptosis inhibitor of macrophage (AIM), which generally associates with IgM in blood and exerts its biological function by dissociation from IgM, may serve as an effective biomarker for NASH-HCC. Here, we established a fully automatic and high-throughput electrochemiluminescence immunoassay (ECLIA) to measure IgM-free AIM and investigated its efficacy in diagnosing NASH-HCC and viral HCC. Methods IgM-free AIM levels were measured in 212 serum samples from patients with, or without, HCC related to NASH, hepatitis B virus, and hepatitis C virus, using ECLIA. We also developed an ECLIA for measuring both IgM-free and IgM-bound AIM and investigated the existing form of AIM in blood by size-exclusion chromatography. Results IgM-free AIM levels were significantly higher in the HCC group than in the non-HCC group, regardless of the associated pathogenesis. Moreover, the area under the receiver operating curve for IgM-free AIM was greater than that for conventional HCC biomarkers, alpha-fetoprotein or des-γ-carboxy prothrombin, regardless of the cancer stage. ECLIA counts of IgM-free AIM derived from samples fractionated by size-exclusion chromatography were significantly higher in patients with NASH-HCC than in healthy volunteers and in patients with non-alcoholic fatty liver and NASH. Conclusions Serum IgM-free AIM may represent a universal HCC diagnostic marker superior to alpha-fetoprotein or des-γ-carboxy prothrombin. Our newly established ECLIA could contribute to further clinical studies on AIM and in vitro HCC diagnosis.

Seroprevalence of Anti-SARS-CoV-2 Antibodies in COVID-19 Patients in Hyderabad, Sindh

Aim: The world is affected by the severe acute respiratory syndrome coronavirus2 (SARS-CoV-2) pandemic. This virus has emerged as a human pathogen that can cause symptoms ranging from fever to Pneumonia, but it remains asymptomatic or mild. To better understand the virus's ongoing spread, identify those who have been infected, and track the immune response, accurate and robust immunological monitoring and SARS-CoV-2 detection assays are needed. Methods: The estimation of serology tests to assess the presence of antibodies to SARS-CoV-2 in COVID- 19 patients at Asian Institute of Medical Sciences (AIMS) and Isra University & hospital. 1229 patients were selected including males and females with the age being 25 to 65 years living in the territories from 1st August to 30th November 2020. The anti-SARS-CoV-2 test was performed by an electrochemiluminescence immunoassay analyzer. Results: Out of 1229 participants 206 (17%) were positive with COVID-19, and 1023 (83%) were negative. The results further revealed that a higher percentage of positive COVID-19 were detected in males in all age groups as compared to females, and most of them are affected at age of 46-65 years male 40 (24.69%) and female 14(17.5%). Conclusion: The seroprevalence of SARS-COV-2 antibodies has increased in the old age population, which may aid in determining the true number of infected cases. Although the current study is based on a small sample of participants, the findings suggest a study with a larger population to implement stronger and targeted interventions.

Phoenixin as a New Target in the Development of Strategies for Endometriosis Diagnosis and Treatment

Small integral membrane protein 20/phoenixin (SMIM20/PNX) and its receptor GPR173 (G Protein-Coupled Receptor 173) play a role in the regulation of the hypothalamic–pituitary–gonadal axis (HPG). The aim of the study was to determine PNX, FSH, LH, and 17β-estradiol association in women with endometriosis, and the expression of SMIM20/PNX signaling via GPR173. Serum PNX, FSH, LH, and 17β-estradiol concentrations were measured by enzyme and electrochemiluminescence immunoassay. SMIM20/PNX and GPR173 expression in the eutopic and ectopic endometrium was assessed by qPCR and immunohistochemistry. Reduced PNX level, increased LH/FSH ratio and elevated 17β-estradiol concentration were found in patients with endometriosis. No differences in SMIM20 expression were observed between the studied endometria. GPR173 expression was lower in ectopic than in eutopic endometria. SMIM20 expression was mainly restricted to stroma. GPR173 was detected in some eutopic and ectopic stromal cells and in eutopic glandular epithelial cells. Discriminant analysis indicates the diagnostic relevance of PNX and LH/FSH ratio in patients with endometriosis. In women with endometriosis, reduced PNX levels and GPR173 expression may be responsible for HPG axis dysregulation. These new insights may contribute to a better understanding of the pathophysiology of endometriosis and provide the basis for a new strategy for diagnosis and treatment of endometriosis.

Clinical performance of the Roche Elecsys SARS-CoV-2 Antigen fully automated electrochemiluminescence immunoassay

Objective: We assessed the clinical performance of novel Roche Elecsys SARS-CoV-2 Antigen fully automated electrochemiluminescence immunoassay (ECLIA). Design and Methods: We tested 160 subjects, 110 (68.8%) with positive molecular test for SARS-CoV-2 infection in nasopharyngeal samples with Altona Diagnostics RealStar SARS-CoV-2 RT-PCR Kit and Roche Elecsys SARS-CoV-2 Antigen. The local imprecision was validated by analyzing three nasopharyngeal samples with different antigen concentration (1.84, 9.51 and 423.30 TCID50/mL) for 20 consecutive times (intra-assay imprecision) or for 10 consecutive working days (inter-assay imprecision).Results: The local intra- and inter-assay imprecision of Elecsys SARS-CoV-2 Antigen ECLIA was found to be comprised between 2.0-2.0% and 5.8-7.6%, yielding to a total imprecision of 6.2-7.8%. Highly significant correlation was found between Elecsys SARS-CoV-2 Antigen ECLIA and cycle threshold (Ct) values of SARS-CoV-2 S and E genes (both r=-0.91; p<0.001). The area under the curve (AUC), sensitivity and specificity of Elecsys SARS-CoV-2 Antigen ECLIA were 0.83, 0.43 and 1.00 in all samples, 0.99, 0.87 and 0.99 in those with both Ct values <30, as well as 1.00, 1.00 and 0.89 in samples with both Ct values <25. Conclusion: Roche Elecsys SARS-CoV-2 Antigen ECLIA may be a surrogate of molecular testing for identification of super-spreaders.

High levels of osteoprotegerin are associated with coronary artery calcification in patients suspected of a chronic coronary syndrome

Plasma osteoprotegerin (OPG) and vascular smooth muscle cell (VSMC) derived extracellular vesicles (EVs) are important regulators in the process of vascular calcification (VC). In population studies, high levels of OPG are associated with events. In animal studies, however, high OPG levels result in reduction of VC. VSMC-derived EVs are assumed to be responsible for OPG transport and VC but this role has not been studied. For this, we investigated the association between OPG in plasma and circulating EVs with coronary artery calcium (CAC) as surrogate for VC in symptomatic patients. We retrospectively assessed 742 patients undergoing myocardial perfusion imaging (MPI). CAC scores were determined on the MPI-CT images using a previously developed automated algorithm. Levels of OPG were quantified in plasma and two EV-subpopulations (LDL and TEX), using an electrochemiluminescence immunoassay. Circulating levels of OPG were independently associated with CAC scores in plasma; OR 1.39 (95% CI 1.17–1.65), and both EV populations; EV-LDL; OR 1.51 (95% CI 1.27–1.80) and EV-TEX; OR 1.21 (95% CI 1.02–1.42). High levels of OPG in plasma were independently associated with CAC scores in this symptomatic patient cohort. High levels of EV-derived OPG showed the same positive association with CAC scores, suggesting that EV-derived OPG mirrors the same pathophysiological process as plasma OPG.

SARS-CoV-2 Antibody Rapid Tests: Valuable Epidemiological Tools in Challenging Settings

Our study provides a fresh perspective on the possible employment of SARS-CoV-2 LFA antibody tests. We developed an in-depth, large-scale analysis comparing LFA performance to enzyme-linked immunosorbent assay (ELISA) and electrochemiluminescence immunoassay (ECLIA) and evaluating their sensitivity and specificity in identifying COVID-19 patients at different time points from symptom onset.