Journal of immunoassay & immunochemistry最新文献

Background: The rising global burden of breast cancer demands early detection and effective treatment, with a focus on prognostic and predictive markers. The eighth edition of the American Joint Committee on Cancer staging manual introduced a new prognostic staging system to increase the predictive power of the existing anatomical staging system of breast cancer. The current study aimed to establish the correlation between Ki67 expression with molecular subtypes and with the pathological prognostic stage of invasive ductal carcinoma.

Materials and methods: A total of 40 patients were included in the study with samples from 32 modified radical mastectomies and 8 biopsies. Hematoxylin and Eosin staining, histopathological analysis and Ki67 immunostaining were conducted. Descriptive and inferential statistical analyses were performed.

Results: Bloom Richardson Grade II was the predominant histological grade. In Grade II cases, 15 of 24 had a Ki67 labeling index of 26-45%, while 6 exceeded 45% (p = 0.001). Pathological prognostic staging reclassified 27 cases, with 24 (75%) downstaged, 3 (9.38%) upstaged, and 5 (15.63%) retaining their clinical stage.

Conclusions: Ki67 immunohistochemistry is an effective tool for assessing proliferative activity of invasive ductal carcinoma, aiding in pathological prognostic stage stratification and offering insights into tumor biology.

Background: Application of antibodies in therapeutics and diagnostics are growing Continually. Herein, we aimed to find the most qualified immunoglobulin (Ig) chemical preparation method.

Methods: A rabbit was immunized against recombinant SARS-CoV-2 nucleocapsid (NP) and reactive polyclonal antibodies were prepared using the ammonium sulfate (AS), caprylic acid (CA), polyethylene glycol (PEG), and caprylic acid/ammonium sulfate (CA/AS) methods. Different antibody solutions were analyzed by SDS-PAGE and subsequently quantified by ImageJ software for further analysis in terms of Ig purity, Ig recovery, and albumin impurity. Ultimately, the prepared antibodies were assessed via Western blotting and ELISA to evaluate their ability to bind NP.

Results: Prepared Ig solutions via the CA/AS method had the highest Ig purity (followed by CA, PEG, and AS) and lowest albumin impurity (followed by CA, AS, and PEG). The PEG method had the highest recovery followed by AS, CA, and CA/AS methods. Moreover, antibodies prepared via different methods demonstrated comparable binding capacities to NP in ELISA and Western blotting.

Conclusions: CA/AS, closely followed by CA, proved to be the most qualified method for the preparation of Ig yielding the highest Ig purity while the PEG method resulted in the highest Ig recovery rate.

Background: Pneumococcal diseases pose a significant public health concern globally, particularly among young children and the elderly. Vaccination plays a crucial role in their prevention. This study evaluated the functional immune responses to Pneumococcal polysaccharide vaccine serotypes in healthy Indian adults before and after administering a single dose of PPSV23 immunization.

Methods: A total of 125 healthy participants aged 18-65 received the PPSV23 vaccine, and their pre- and post-immunization sera were analyzed by MOPA. Opsonic Index, Geometric mean OPA titers and fold increase for each serotype was calculated.

Results: The highest baseline OPA GMTs were observed for serotypes 33F,17F,9N,20, and 6B. The lowest OPA GMTs were noted against types 1 and 12F. OPA GMTs post-vaccination increased significantly for all serotypes, with geometric mean fold rise (GMFR) ranging from 4.3 to 267.5. Participants with low pre-immunization OPA titers (<8 & <64) showed significant increases in OI fold raise across all tested serotypes post-vaccination. This robust immune response was consistent across serotypes, indicating highly effective seroconversion in individuals with low baseline antibody levels.

Conclusion: The PPSV23 vaccine elicits a strong immunogenic response in individuals with low pre-immunization OPA titers, achieving substantial increases in opsonic index fold raise across various serotypes.

Vitiligo is a common skin disorder where melanocytes, the cells that produce skin pigment, are destroyed by the immune system, leading to white patches on the skin and mucous membranes. This condition affects 0.4% to 2.0% of the global population, with a higher prevalence in females and often beginning in childhood. In India, about 1% of the population is affected, particularly in northern regions, with a higher incidence in females and links to other autoimmune diseases. This review examines recent progress in understanding vitiligo and its treatment. It focuses on the genetic, autoimmune, and environmental factors involved in the disease and highlights new therapies, such as targeted molecular treatments and advanced repigmentation methods. Current research shows that oxidative stress and genetic predispositions contribute to the autoimmune destruction of melanocytes. Novel drug delivery systems, including liposomes, nanoemulsions, and nanostructured lipid carriers, have improved treatment effectiveness. Clinical trials are exploring new treatments like Ruxolitinib cream and melanocyte transplantation, while teledermatology is becoming useful for managing patients. Vitiligo also poses a significant economic burden due to its impact on patients' quality of life. Continued research is essential to develop better, more accessible treatments and reduce the economic impact of vitiligo.

Our study evaluated the possible interference of Burosumab (human recombinant monoclonal antibody directed against N-terminal domain of FGF23) on the immunoassay of intact FGF23 (iFGF23) with the Liaison XL. The analytical method uses three different antibodies, one of which directed against the N-terminal portion of FGF23. The evaluation of the method accuracy involved the fully automated execution of a dilution test on EDTA plasma from 5 subjects who had not received any monoclonal antibody (mAb), 20 EDTA plasma from patients treated with Burosumab, and 2 EDTA plasma from subjects who had not received any mAb in witch an adequate volume of Burosumab had been added in vitro. One sample with specific diluent (LIAISON® FGF 23) with an adequate volume of Burosumab had been added in vitro. The dilution assay provided highly inaccurate iFGF23 results in samples with therapeutic concentrations of Burosumab and in samples with concentrations below the LoQ (6.5 pg/mL). The addition of Burosumab to the diluent did not produce any analytical interference. Dissociation of iFGF23 from the mAb-target complex in diluted sample could explain the loss of accuracy in the iFGF23 immunoassay using the Liaison XL analyzer. Burosumab could be an interferent in immunoassay procedures of iFGF23.

Aim: In 2019, cardiovascular diseases accounted for 32% of global deaths. So, early detection of cardiac disorders is crucial. The study aimed to examine the suitability of Pro-neurotensin and Heart-type fatty acid binding protein as dependable biomarkers for cardiac patients with Heart failure as a primary diagnosis.

Methodology: The prospective study involved 204 Egyptian volunteers (100 cardiac patients and 104 controls) enrolled from El-Sahel Teaching Hospital in Cairo, Egypt, between October 2022 and May 2023. Inclusion criteria included a high risk of cardiovascular events with symptoms like a fast or irregular pulse, shortness of breath, and fatigue. Exclusion criteria included asymptomatic individuals, cognitive disorders, and psychiatric conditions. The Research Ethics Committee approved the protocol. The consultant conducted a clinical examination of all patients and assessed their heart state. Serum ProNT and H-FABP were detected using a kit for the sandwich ELISA technique.

Results: ProNT and H-FABP were significantly elevated in patients compared to controls with p < 0.001. Demonstrated sensitivity of 81% and 84%, with a specificity of 89% and 91%, respectively.

Conclusion: Elevated ProNT and H-FABP levels are associated with severe CVDs, suggesting their potential as diagnostic biomarkers for patients, specifically those with heart failure, as a primary characteristic.

Rotavirus diarrhea and Ascaris lumbricoides (Al) infection increase intestinal morbidity and were associated with altered immune responses that compromise the vaccine efficacy in children. The serum level of rotavirus specific IgA (RV-IgA) and cytokine profiles in A. lumbricoides (AI) infected preschool-aged Nigerian children were estimated following oral rotavirus vaccination. Nineteen of the 149 preschool-aged children (aged 6 to 60 months) with Ascaris lumbricoides infection paired with age and sex-matched helminth - free children were administered with oral rotavirus vaccine after intestinal helminth screening using stool sample concentration technique. Separated sera from 3 mL venous blood samples were collected and estimated for cytokines (IFN-γ, TNF-α, IL-4, IL-8 IL-6, IL-10) and RV-IgA before and three weeks after rotavirus vaccination using Enzyme Linked Immunosorbent Assay. IFN-γ, IL-8, IL-4 were significantly lower at post-vaccination in Al-infected children compared with pre-vaccination. Serum IL-10 was significantly higher at post-vaccination in both Al-infected children and helminth-free controls, compared with pre-vaccination levels (p < 0.05). Pre-vaccination IL-8 and IL-6 were significantly higher in Ascaris lumbricoides-infected children, while the post-vaccination IL-8 was significantly higher in Ascaris lumbricoides-infected compared with control. At post-vaccination period, RV-IgA level was lower in Al-infected children and significantly higher in helminth - free control group compared to pre-vaccination RV-IgA level. Ascaris lumbricoides infection contributed to down-regulation of some cytokines and antibody responses to oral rotavirus vaccine.

Background: Numerous genes are involved in immune system modulation, and their polymorphisms may contribute to developing autoimmune disorders. Genetic variation contributes significantly to disease susceptibility to autoimmune thyroid disease (AITD).

Objectives: This work aims to investigate the role of single-nucleotide polymorphisms (SNPs) of interferon induced with helicase C domain 1 (IFIH1) rs1990760 and glutamate decarboxylase (GAD) rs769404 in AITD development.

Methods: The study had 330 participants, including 153 cases of Hashimoto's thyroiditis (HT), 77 cases of Graves' disease (GD), and 100 healthy controls. All subjects underwent medical history assessment and clinical evaluation. Tests were conducted using real-time PCR, including genotyping of IFIH1 (rs1990760) and GAD (rs769404) via an allele discrimination assay.

Results: Most patients with AITD were females. About 18.3% of HT cases and 15.6% of GD cases have a positive family history of thyroid disease. A significant statistical difference was observed between AITD cases and control regarding IFIH1 (rs1990760) and GAD (rs769404) gene polymorphism. Moreover, GD patients, HT patients, and the control group showed increased CT and TT alleles in patients compared to those in controls.

Conclusion: IFIH1 and GAD polymorphisms are involved in AITDs (HT and GD) development and are associated with some clinical presentations. HT and GD cases had a positive family history of thyroid disease. There was a significant statistical difference between AITD cases and control regarding IFIH1 (rs1990760) and GAD (rs769404) gene polymorphism.

This study is aimed at determining the prevalence of oral HPV infection and the risk indicators for oral HPV carriage in people living with HIV. Data on socio-demographics, sexual behavioral practices, and lifestyle practices of the participants were collected from 66 people living with HIV. The HIV parameters of each study participant were obtained from clinical records. Oral rinses obtained from each participant were subjected to HPV ELISA antigen test for screening and extracted DNA was subjected to nested PCR and whole-genome sequencing for genotyping. Approximately 36% (10 of 28) HIV-positive individuals had oral HPV carriage with one person carrying oncogenic type, HPV16. In addition, 80% (8 of 10) of those with HPV positivity by PCR are females, but with no statistically significant association. The CD4 count showed no significant association with oral HPV carriage in HIV positive individuals; however, age at first sex is a determinant of oral HPV infection in people living with HIV with positive association observed on both bivariate analysis and logistic regression (AOR: 150.49, 95% CI: 1.40-16,155.47, p = 0.036).