Journal of immunoassay & immunochemistry最新文献

Background: IL2 is one of the key cytokines essential for regulating the immune system and the inflammation-related carcinogenesis process. Few studies have examined the relationship between lung cancer and the IL2-330 (rs2069762) gene polymorphism, despite several studies demonstrating that it is linked to numerous cancer types.

Objective: Our study aimed to investigate the association between IL2-330 (rs2069762) polymorphism and lung cancer risk and explore the role of IL2-330 polymorphism in survival outcomes (OS and PFS).

Patients and methods: The study was conducted from October 2023 to November 2024, including 50 randomly selected patients diagnosed with lung cancer and 50 subjects matched for age and gender were used as controls in this case-control study. The IL2-330 (rs2069762) gene polymorphism was assessed using real-time PCR.

Results: We found that the AC genotype was associated with a notably lower risk of lung cancer in comparison to the AA genotype (95% CI: 0.06-0.61, p = 0.01). Conversely, the CC genotype showed no significant association with lung cancer risk when compared to the reference genotype. The comprehensive comparison of survival distributions among the AA, AC, and CC genotypes through the Log-Rank (Mantel-Cox) test indicated no statistically significant difference.

Conclusions: According to our research, The AC genotype of IL2-330 (rs2069762) is associated with a significantly higher survival rate and lower risk of lung cancer. Further future studies are needed to confirm these findings.

Oral squamous cell carcinoma (OSCC) accounts for over 90% of all oral malignancies and has a 50-60% five-year survival rate, despite advances in treatment. Ki67, a nuclear protein involved in cellular proliferation, has been studied as a prognostic marker in various malignancies, including OSCC. However, its link with different histological grades of OSCC is uncertain. The study aimed to evaluate Ki-67 expression in well-differentiated (WDOSCC), moderately differentiated (MDOSCC), and poorly differentiated (PDOSCC) tumors to ascertain its correlation with tumor differentiation and aggressiveness. Forty OSCC cases were classified using Broder's histological grading system. Ki67 immunohistochemical staining was performed, and three groups were categorized based on Ki67 expression: low (1-25%), moderate (26-50%), and high ( > 50%) proliferation. Statistical analysis assessed the significance of Ki67 expression across OSCC grades. Among 40 cases, 35% were WDOSCC, 45% MDOSCC, and 20% PDOSCC. Low Ki67 proliferation was seen in 64% of WDOSCC, moderate in 61% of MDOSCC, and high in 75% of PDOSCC. There was a significant (p < 0.001) association between OSCC grading and Ki67 expression. Ki67 expression correlates with OSCC histological grades, increasing with tumor proliferation. These findings support Ki67 as a prognostic marker, warranting further large-scale validation studies.

Background: Diagnosis is an important factor in controlling disease. Single-chain fragment variables (scFvs) can be used for diagnosis; however, due to their immobilization issues, their application has been limited. Herein, we isolated a SARS-CoV-2 nucleocapsid phosphoprotein (NP)-specific scFv and propose it as a diagnostic tool in the scFv-displaying phage format to overcome the immobilization issue.

Method: Spleen from NP-immunized BALB/c mice was isolated, total RNA was extracted, and cDNA was synthesized. An scFv library was constructed, using the splicing by overlap extension (SOE) PCR technique, which was cloned into the pCANTAB5E phagemid. The phage library was panned against the NP antigen, and the output phages with the highest binding capability were screened for the most qualified scFv, which was later assessed in terms of sensitivity and specificity.

Results: The scFv-displaying phage library was panned against the recombinant NP in three rounds and 40 randomly selected colonies from the third round's outputs were screened. Alongside several clones, clone #31 was chosen as the most qualified scFv, which later exhibited favorable sensitivity and specificity against NP in further ELISA-based experiments.

Conclusions: Clone #31 could be utilized to develop diagnostic tools and therapeutics against SARS-CoV-2.

Chronic Obstructive Pulmonary Disease is a major global health concern with significant morbidity and mortality, characterized by heterogeneity influenced by inflammation, oxidative stress, and autoimmunity. This study investigated the role of autoimmunity in stable and exacerbated COPD phenotypes using proteomic and immunological analyses to elucidate molecular mechanisms and identify potential biomarkers. Plasma samples from COPD phenotypes and healthy controls were analyzed using label-free mass spectrometry to identify differentially expressed proteins. Functional annotation and pathway enrichment analysis highlighted proteins linked to autoimmunity, while immunological assays assessed anti-nuclear antibody prevalence and intensity using ANA ELISA and indirect immunofluorescence. The study identified differentially expressed proteins, namely, DNA repair protein XRCC2, phosphatidyl inositol glycan-specific phospholipase D, E3 ubiquitin protein ligase SHPRH, and Protocadherin-β, implicated in autoimmune pathways. Pathway enrichment analysis of these proteins highlighted the uPAR-mediated signaling, mTOR, PI3K/Akt, ARF6, and S1P signaling pathways, known for their roles in autoimmunity. Immunological assays revealed ANA positivity in 47% of stable COPD and 36% of exacerbated COPD, among which, 80% exhibited a speckled fluorescence pattern, often associated with anti-SSA and anti-SSB antibodies. The findings highlight the potential role of autoimmunity in COPD pathogenesis, suggesting phenotype-specific immune dysregulation, providing a basis for future biomarker and therapeutic research.

This study explores the relationship between oxidative - antioxidative balance and stromal cell-derived factor-4 (SDF-4) levels in individuals with Generalized Anxiety Disorder (GAD), aiming to clarify their roles in the disorder's pathophysiology. By examining these biomarkers, the research investigates whether oxidative stress mechanisms contribute to GAD. A cross-sectional study was conducted with 43 GAD patients and 40 age- and sex-matched healthy controls. Participants were assessed using the Beck Anxiety Inventory (BAI) and the Structured Clinical Interview for DSM-5. Serum levels of SDF-4, total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) were measured. GAD patients showed significantly higher levels of SDF-4, TAS, TOS, and OSI compared to controls (p < 0.001). No significant differences were found in other hematological parameters. In the GAD group, SDF-4 levels were not significantly correlated with anxiety severity or oxidative markers. ROC analysis indicated good diagnostic performance of SDF-4 for GAD (AUC = 0.870). The findings suggest that oxidative - antioxidative imbalance and elevated SDF-4 levels may play a role in GAD. SDF-4 might represent a compensatory response to oxidative stress. Elevated TAS levels may indicate enhanced antioxidant defense. Further studies are needed to clarify the mechanistic and therapeutic implications.

Serum-free light-chain assays are important in the diagnosis and in monitoring therapeutic responses of plasma cell disorders and are complementary to serum protein electrophoresis. The serum-free light-chain assay detects the light-chain portion of immunoglobulin in its free form with high sensitivity. In combination with serum protein electrophoresis and serum immunofixation electrophoresis, the free light-chain assay serves an important role in predicting disease progression in monoclonal gammopathy. Here, we compare the performance of a cartridge-based system Mispa i3 using Diazyme reagent in comparison to Roche Cobas using Freelite reagent. Both of these reagents use polyclonal antibodies for the detection of serum-free light chains. Mispa i3 is a cartridge-based protein analyzer and has a unique channel shifting technology with both turbidimetric and nephelometric principles for immuno assays. Samples of 196 patients were included in this study, and very good agreement was observed between these two assays. Our data show that even though discrepancies were observed for high concentration samples, they are clinically correlated by the free light-chain ratios. We observed a very good concordance of 89% between these two assays for free light-chain ratios.

Delta-9-tetrahydrocannabinol (THC) is one of the cannabinoid metabolites present in Cannabis sativa. Currently, cannabis is an ingredient in foods, cosmetics, and medicinal products. Many countries control cannabis and THC for illegal or under-regulation products. This study focused on development of recombinant antibodies specific to THC, and their application of analytical methods using enzyme-linked immunosorbent assay (ELISA) techniques. Conjugation of THC with bovine serum albumin or ovalbumin was performed using Mannich and N,N'-carbonyldiimidazole reactions. The recombinant antibody was expressed from Escherichia coli in form of fragmented antigen-binding (Fab) antibody. Antigen-conjugates and Fab were performed using ELISA-based analytical method and validated method for sensitivity, accuracy, and precision. The developed Fab in this study demonstrates high specificity for detection of THC. The analytical range was between 0.78 and 25 µg/mL, with limit of detection of 0.59 µg/mL. Accuracy ranged from 98.15-108.97%, with intra- and inter-assay % relative standard deviation of 2.15-8.03 and 1.83-8.45%, respectively. Parameters for method validation, including accuracy and precision, were within acceptable limits which are comparable to modified HPLC method from previous study. The approach demonstrated outstanding sensitivity and specificity for analysis of THC in cannabis extracts, suggesting it to be suitable for the quantitative assessment of THC in variety of products.

Immunohistochemistry (IHC) using the BRAF V600E antibody is a sensitive and specific method for detecting BRAF V600E mutations in colorectal cancer (CRC). Given that BRAF and KRAS mutations are mutually exclusive, this study aimed to assess the expression of BRAF V600E in CRC according to KRAS mutation status. Automated IHC analysis was performed on tissue samples from metastatic CRC patients diagnosed between 2012 and 2022 using the anti-BRAF V600E antibody (GenomeME, IHC600). Positive and negative control tissues were used for validation. Cytoplasmic staining was considered positive, with intensity classified as weak, moderate, or strong. The percentage of positive tumor cells was recorded semi-quantitatively. Thirty-five cases were included.The mean age of patients was 60 years (±12.41), with a male-to-female ratio of 2.9. KRAS mutations were present in 48% of cases. BRAF V600E cytoplasmic staining was observed in 37.1%, with a mean percentage of positive cells of 50% (range: 5%-100%). Diffuse and focal staining were found in 7/13 and 6/13 cases, respectively. Significant associations were observed with mucinous carcinoma subtype, invasion patterns, and lymph node metastasis. All KRAS-mutated cases showed negative BRAF staining. Complete absence of BRAF V600E IHC staining in CRC is strongly associated with KRAS mutation, suggesting IHC as an efficient tool to predict KRAS mutational status.

Avian influenza is one of the major threats to poultry and human health in northern Benin, while mixed-species farming systems increase the risks of viral transmission. The present study estimated the seroprevalence of avian influenza subtypes H9N2 and H5 in indigenous chickens and guinea fowls in the Atacora and Donga regions. A total of 300 birds including 191 indigenous chickens and 109 guinea fowls, from six districts were sampled through a cross-sectional survey using systematic random sampling. Hemagglutination inhibition assay was used to detect antibodies, revealing an overall H9N2 seroprevalence of 41%, with 17.5% of samples testing positive for H5. The seroprevalence of H9N2 was notably higher in guinea fowls (51.81% in Atacora and 52% in Donga) compared to chickens (34.95% in Atacora and 34.83% in Donga). H5 antibodies were found only in guinea fowls in Atacora (46.66%). The study also found that farms with both chickens and guinea fowls had a significantly higher odds ratio for H9N2 positivity (OR = 4.25, p < 0.001) compared to chicken-only farms. The results underscore the importance of mixed-species systems in the transmission of avian influenza, suggesting that targeted surveillance and biosecurity measures are essential for controlling the spread of these viruses.

Background: Recent studies have reported aberrant GATA3 immunohistochemical expression in high-grade prostate carcinoma, raising concerns about its specificity. This study aimed to evaluate GATA3 immunohistochemical expression in prostate adenocarcinoma.

Methods: We retrospectively collected cases of primary prostate adenocarcinoma diagnosed in our pathology department(2019-2024). Clinical and pathological data, including Gleason and ISUP grades, were collected.An automated immunohistochemical analysis was performed using the anti-GATA3 antibody(Leica, L50-823). Only nuclear staining was considered positive.

Results: Fifty-eight cases have been included in the study with a mean age of 67.8 years-old. GATA3 nuclear expression was observed in 6.9% (4/58) of cases, all classified as Gleason 7 (4   +   3) and ISUP grade 3. Only the Gleason 4 or 5 components showed staining, while Gleason 3 components were negative. Additionally, benign prostate glands and seminal vesicles showed rare GATA3 expression (4/58 and 1/58 cases, respectively).

Conclusion: While GATA3 is commonly used as a marker for urothelial carcinoma, our findings indicate that it can also be expressed in prostate adenocarcinoma, particularly in higher Gleason grade components, which may pose a diagnostic pitfall. Further studies are needed to determine the clinical significance of GATA3 expression in prostate adenocarcinoma and its potential impact on tumor behavior and prognosis.