Fluorescence Immunoassay Research Articles

Comparative analysis of contemporary anti-double stranded DNA antibody assays for systemic lupus erythematosus.

Elevated double-stranded DNA (dsDNA) antibody levels in blood serum are considered a disease-specific marker in systemic lupus erythematosus (SLE), correlate with disease activity and the incidence of lupus nephritis, and can be detected in up to 86% of all SLE cases. Despite the high clinical relevance, the variety of dsDNA antibody testing methods with heterogenous performance in clinical use remains challenging. This study is the first to prospectively investigate the performance of two of today's most commonly applied anti-dsDNA testing methods head-to-head under real-world conditions, as well as their correlation with other clinical and serological disease parameters in SLE patients. In this prospective study, all SLE patients undergoing treatment at the Department of Rheumatology at the University Hospital Bonn within a 13-months period (n=41) and control patients without connective-tissue disease (n=51) were consecutively enrolled and examined. For all study participants' serum samples both anti-dsDNA-NcX enzyme-linked immunoassay testing EUROIMMUN, Luebeck, Germany) and the fluorescence immunoassay ELiA dsDNA (Thermo Fisher Scientific, Waltham, USA) were performed. In addition, demographic data, further laboratory values and disease activity parameters were recorded. Clinical disease activity was assessed by SLEDAI-2K. Both assays showed high specificity (anti-dsDNA-NcX ELISA: 0.9, ELiA dsDNA: 0.959), but there were notable differences in sensitivity (anti-dsDNA-NcX ELISA: 0.51, ELiA dsDNA: 0.38). Pearsons's correlation yielded a positive correlation between anti-dsDNA concentrations and CRP concentrations for the anti-dsDNA-NcX ELISA (R=0.22; p=0.038) and a mild-to-moderate inverse correlation between concentrations of anti-dsDNA and complement C4 for the ELiA dsDNA test (R=-0.22; p=0.045) when SLE and control patients were considered together. Other than, no significant correlation between anti-dsDNA concentrations and clinical or laboratory findings was found for either test procedure. Both anti-dsDNA antibody assays represent reliable examination methods with high specificity for the diagnosis of SLE that fulfill EULAR/ACR requirements. However, the anti-dsDNA-NcX ELISA showed superior sensitivity and significant correlation with disease activity (as measured by CRP concentrations).

Cancer Marker Immunosensing through Surface-Enhanced Photoluminescence on Nanostructured Silver Substrates.

Nanostructured noble metal surfaces enhance the photoluminescence emitted by fluorescent molecules, permitting the development of highly sensitive fluorescence immunoassays. To this end, surfaces with silicon nanowires decorated with silver nanoparticles in the form of dendrites or aggregates were evaluated as substrates for the immunochemical detection of two ovarian cancer indicators, carbohydrate antigen 125 (CA125) and human epididymis protein 4 (HE4). The substrates were prepared by metal-enhanced chemical etching of silicon wafers to create, in one step, silicon nanowires and silver nanoparticles on top of them. For both analytes, non-competitive immunoassays were developed using pairs of highly specific monoclonal antibodies, one for analyte capture on the substrate and the other for detection. In order to facilitate the identification of the immunocomplexes through a reaction with streptavidin labeled with Rhodamine Red-X, the detection antibodies were biotinylated. An in-house-developed optical set-up was used for photoluminescence signal measurements after assay completion. The detection limits achieved were 2.5 U/mL and 3.12 pM for CA125 and HE4, respectively, with linear dynamic ranges extending up to 500 U/mL for CA125 and up to 500 pM for HE4, covering the concentration ranges of both healthy and ovarian cancer patients. Thus, the proposed method could be implemented for the early diagnosis and/or prognosis and monitoring of ovarian cancer.

Time-resolved fluorescence immunoassay based on glucose oxidase-encapsulated metal-organic framework for amplified detection of foodborne pathogen

BackgroundFluorescence immunoassays are commonly employed for the detection of pathogenic bacteria as a means of ensuring food safety and preserving public health. However, the challenges such as poor photostability and background interference have limited their sensitivity and accuracy. The emergence of metal-organic frameworks (MOFs) as a label probe offers a promising solution for advancing fluorescence immunoassays owing to their tunable nature. Nonetheless, the low fluorescence efficiency of MOFs and the potential risk of dye leakage pose obstacles to achieving high detection sensitivity. Therefore, there exists a pressing need to fully utilize the potential of MOF composites in fluorescence immunoassays. ResultsWe explored the potential of glucose oxidase-encapsulated zeolitic imidazole framework-90 (GOx@ZIF-90) as a label probe to construct a time-resolved fluorescence immunoassay with amplified detection signal. This immunoassay involved functionalizing Fe3O4 nanoparticle with porcine antibody to specifically capture and separate the target bacteria, Staphylococcus aureus (S. aureus). The captured S. aureus was then bound by GOx@ZIF-90 modified with vancomycin, resulting in a fluorescence response in the europium tetracycline (EuTc). The encapsulation of GOx in ZIF-90 provided a confinement effect that significantly enhanced the catalytic activity and stability of GOx. This led to a highly efficient conversion of glucose to H2O2, amplifying the fluorescence signal of EuTc. The immunoassay demonstrated a high sensitivity in detecting S. aureus, with a detection limit of 2 CFU/mL. We also obtained satisfactory results in milk samples. Attractively, the time-resolved detection mode of EuTc allowed the immunoassay to eliminate background fluorescence and enhance accuracy. SignificanceThis study not only presented a new method for detecting foodborne pathogens but also highlighted the potential of enzyme-encapsulated MOF composites as label probes in immunoassays, providing valuable insights for the design and fabrication of MOF composites for various applications.

Analysis on epidemiological characteristics of allergens with allergic skin diseases in a hospital in Beijing City from 2017 to 2021

Objectives: In order to provide valuable information for the diagnosis and treatment of dermatitis, the prevalence rate and trend changes of common allergens in patients with allergic skin diseases in Beijing City were investigated and analyzed. Methods: This study was a retrospective data collection study. A total of 2 822 patients diagnosed with moderate to severe atopic dermatitis with allergen examinations were collected from Peking University First Hospital from 2017 to 2021, most of them were adults, and 83%(2 340/2 822) were over 18 years old. The ImmunoCAP fluorescent enzyme-linked immunoassay system was used to quantify specific IgE. The positive rate of each allergen was counted according to age, gender and year. The epidemiological characteristics and trends were analyzed. Results: In the past five years, 22 503 allergen-specific IgE test data were collected, and the positive rate of allergens in 40%(1 122/2 822) of the 2 822 patients was mainly multiple sensitization, the positive rate of allergens and multiple sensitization rates were the highest in people aged 13-18 years old. The 2 822 patients were dominated by inhaled allergens, and the highest positive rate was inhaled mixture (29.3%). The top five positive rates of inhaled allergens were Dermatophagoides farina (26.7%), Dermatophagoides pteronyssinus (25.5%), Mugwort (23.9%), Birch pollen (19.2%) and Siberian cocklebur (18.2%). The top five positive rates of food allergens were shrimp (15.2%), peanut (14.8%), sesame (14.7%), wheat (13.2%) and milk (13.1%). Analysis of the positive rate of allergens in different age groups showed except for the positive rate of birch pollen peaked at 3-12 years old, other inhaled allergens were the highest food allergens in 13-18 years old. Among food allergens, the positive rates of fx5, milk, egg and wheat were the highest in <3 years old, the positive rate decreased gradually with age, and the positive rate of other food allergens peaked at 13-18 years old, and the shrimp and crab was the highest in adult. The positive rate of various allergens in the 2 822 patients showed little difference between male and female and the male was slightly higher than female. In the past five years, the positive rate of allergens has been on the rise. The changes of inhaled allergens in cat/dog hair dander, Dermatophagoides farina and cicadas were large, while the positive rate of food allergens was stable. Conclusions: The etiology of the allergic skin diseases is complex, which may be related to allergen exposure, heredity, immunity, environmental factors, abnormal skin barrier function, etc. This study showed the distribution of allergens in patients with allergic skin diseases in Beijing City to a certain extent. It provided epidemiological data and clinical evidence for the prevention and treatment of allergic diseases.

Ratiometric fluorescence and absorbance dual-model immunoassay based on 2,3-diaminophenazine and carbon dots for detecting Aflatoxin B1

In this work, a dual-model immunoassay for detecting Aflatoxin B1 (AFB1) was developed based on 2,3-diaminophenazine (DAP) and carbon dots (CDs). Under the catalysis of horseradish peroxidase (HRP), the o-phthalylenediamine (OPD) was oxidized to DAP which had a yellow color and intense fluorescence. The color changes form colorless to yellow was used to design absorbance model immunoassay. Meanwhile, the absorption spectrum of DAP overlapped with the emission spectrum of CDs which caused the fluorescence of CDs to be quenched. The fluorescence changes of DAP and CDs were used to develop ratiometric fluorescence immunoassay. The dual-model immunoassay showed excellent sensitivity with the limits of detection (LODs) of 0.013 ng/mL for fluorescence mode and 0.062 ng/mL for absorbance mode. Meanwhile, both models exhibited great selectivity for AFB1. Additionally, the recovery rates suggested the proposed dual-model immunoassay had great potential in actual samples detection.

Effects of different inhalant allergens on lung functions in adult patients with bronchial asthma.

To analyze the effects of different types of inhalant allergens on the lung functions of adult patients with bronchial asthma. This cross-sectional study included a total of 47 adults diagnosed with bronchial asthma at the Respiratory Outpatient Department of Tianjin First Central Hospital. Patients were divided into non-sensitized and sensitized groups based on the number of positive allergens detected and classified into four groups (the dust mite mixed group, animal dander mixed group, pollen-mixed group, and mold mixed group) based on the type of positive allergen detected. They were tested for the serum concentration of allergen-specific immunoglobulin E (sIgE) using a fluorescence immunoassay analyzer, and lung function was assessed using a pulmonary function testing machine. One-way analysis of variance was used to compare normally distributed data, while the rank sum test was utilized for non-normally distributed data. There was no statistically significant difference in lung function indicators between these two groups (p > .05). There were statistically significant differences in forced expiratory volume in one second as a percentage of the predicted value (FEV1 %pred) (p = .028), FEV1 /forced vital capacity as a percentage of the predicted value; (FVC%pred) (p = .016), peak expiratory flow as a percentage of the predicted value (PEF%pred) (p = .001), forced expiratory flow at 50% of the predicted value of forced vital capacity (FEF50%pred) (p = .003), forced expiratory flow at 75% of the predicted value of forced vital capacity (FEF75%pred) (p = .023), and maximal midexpiratory flow (MM)EF75/25%pred (p = .002) among the four groups. The pollen-mixed group had higher PEF%pred (pollen vs. animal dander, p = .067; pollen vs. dust-mites, p = .008; pollen vs. molds, p = .001) and MMEF75/25%pred (pollen vs. animal dander, p = .048; pollen vs. dust-mites, p = .003; pollen vs. molds, p = .001) than the other three groups. The pollen-mixed group had higher FEF50%pred than the dust-mites mixed group (p = .008) and molds-mixed group (p = .001). The pollen-mixed group had higher FEF75%pred (p = .005), FEV1 %pred (p = .001), and FEV1 /FVC%pred (p = .001) than the molds-mixed group. Different inhalant allergens had different effects on lung functions in adults with asthma.

EE315 The Budget Impact of Including the Automated Time-Resolved Fluorescence Immunoassay As Point-of-Care Testing on Cardiac Markers in an Emergency Department in China

EE315 The Budget Impact of Including the Automated Time-Resolved Fluorescence Immunoassay As Point-of-Care Testing on Cardiac Markers in an Emergency Department in China

In Situ Real-Time Monitoring for Aseptic Drilling: Lessons Learned from the Atacama Rover Astrobiology Drilling Studies Contamination Control Strategy and Implementation and Application to the Icebreaker Mars Life Detection Mission.

In 2019, the Atacama Rover Astrobiology Drilling Studies (ARADS) project field-tested an autonomous rover-mounted robotic drill prototype for a 6-Sol life detection mission to Mars (Icebreaker). ARADS drilled Mars-like materials in the Atacama Desert (Chile), one of the most life-diminished regions on Earth, where mitigating contamination transfer into life-detection instruments becomes critical. Our Contamination Control Strategy and Implementation (CCSI) for the Sample Handling and Transfer System (SHTS) hardware (drill, scoop and funnels) included out-of-simulation protocol testing (out-of-sim) for hardware decontamination and verification during the 6-Sol simulation (in-sim). The most effective five-step decontamination combined safer-to-use sterilants (3%_hydrogen-peroxide-activated 5%_sodium-hypochlorite), and in situ real-time verification by adenosine triphosphate (ATP) and Signs of Life Detector (SOLID) Fluorescence Immunoassay for characterization hardware bioburden and airborne contaminants. The 20- to 40-min protocol enabled a 4-log bioburden reduction down to <0.1 fmoles ATP detection limit (funnels and drill) to 0.2-0.7 fmoles (scoop) of total ATP. The (post-cleaning) hardware background was 0.3 to 1-2 attomoles ATP/cm2 (cleanliness benchmark background values) equivalent to ca. 1-10 colony forming unit (CFU)/cm2. Further, 60-100% of the in-sim hardware background was ≤3-4 bacterial cells/cm2, the threshold limit for Class <7 aseptic operations. Across the six Sols, the flux of airborne contaminants to the drill sites was ∼5 and ∼22 amoles ATP/(cm2·day), accounting for an unexpectedly high Fluorescence Intensity (FI) signal (FI: ∼6000) against aquatic cyanobacteria, but negligible anthropogenic contribution. The SOLID immunoassay also detected microorganisms from multiple habitats across the Atacama Desert (anoxic, alkaline/acidic microenvironments in halite fields, playas, and alluvial fans) in both airborne and post-cleaning hardware background. Finally, the hardware ATP background was 40-250 times lower than the ATP in cores. Similarly, the FI peaks (FImax) against the microbial taxa and molecular biomarkers detected in the post-cleaned hardware (FI: ∼1500-1600) were 5-10 times lower than biomarkers in drilled sediments, excluding significant interference with putative biomarker found in cores. Similar protocols enable the acquisition of contamination-free materials for ultra-sensitive instruments analysis and the integrity of scientific results. Their application can augment our scientific knowledge of the distribution of cryptic life on Mars-like grounds and support life-detection robotic and human-operated missions to Mars.

CD22 CAR T Cell-Related IEC-HS Is Associated with an IFN-γ Cytokine Signature

Introduction Chimeric antigen receptor T-cell therapy targeting CD22 (CAR22) shows promise for patients with relapsed / refractory B-cell acute lymphoblastic leukemia (ALL) and large B cell lymphoma (LBCL) (Fry et al. Nat Med 2018; Baird et al. Blood 2020). The overall response rate for CAR22 in adults who received prior CD19 CAR therapy is 100% and 68% in ALL and LBCL respectively (Tandem 2023) (NCT04088890). However, 33% of pediatric ALL patients who received CAR22 developed immune effector cell-associated hemophagocytic lymphohistiocytosis-like syndrome (IEC-HS) (Shah et al. JCO 2020). IEC-HS is characterized by hyperferritinemia, cytopenias, coagulopathy and end organ damage. Pediatric ALL data suggest many cytokines including IFN-γ, IL-1β, IL-18 and TNF-α are upregulated in IEC-HS (Ishii et al. JCI 2020, Lichtenstein et al. Blood 2021). Here, we report the clinical features and cytokines associated with IEC-HS after CAR22 infusion in predominantly adult subjects with ALL and LBCL. Methods Plasma was collected weekly from Day 0 to 28 for 54 patients (ALL: n = 16, LBCL: n = 38; adult: n = 51, pediatrics: n = 3) who were treated with CAR22 manufactured using an automated, closed-system Miltenyi CliniMACS Prodigy device. Three dose levels were tested (ALL: 3 x 10 5 cells/kg, LBCL dose level 1 (DL1): 1 x 10 6 cells/kg, LBCL dose level 2 (DL2): 3 x 10 6 cells/kg). Clinical characteristics associated with IEC-HS were retrospectively collected and graded according to consensus criteria (Hines et al. TCT 2023). Eighty cytokines were quantified in batch via multiplexed fluorescence immunoassay (Luminex) to generate mean fluorescent indices (MFIs). Unsupervised hierarchical clustering using Euclidean distance and ward.D2 linkage were employed to analyze cytokine dynamics. Statistical analyses were performed using R (v 4.2.2). Results 8 of 54 (15%) patients developed IEC-HS, all were ≥18 years old. 5 patients had LBCL, 3 had ALL. 5 cases were grade 2, 3 were grade 4. Median onset was D+12 (range 8 - 21). All patients had prior CRS with median onset at D+1 (range 0 - 9). The 3 ALL patients had ongoing CRS and spectral overlap with IEC-HS at time of diagnosis whereas LBCL patients' CRS resolved prior to IEC-HS onset. IEC-HS patients had significant increases in inflammatory markers (ferritin, CRP, LDH, triglycerides), coagulation markers (INR, PTT), cytopenias (hemoglobin, platelets) and transaminases (Table 1). Patients most commonly received high-dose glucocorticoids, tocilizumab and anakinra. Additional agents for grade 4 IEC-HS included anti-thymoglobulin, dasatanib and emapalumab. 5 of 8 patients developed serious infections requiring IV antimicrobials including bacteremia, invasive fungal infections and viral reactivation. 5 patients with IEC-HS died after achieving complete response: 3 due to IEC-HS and concomitant infection, 1 from t-AML and 1 lost to follow up. IEC-HS occurred more frequently at higher doses in LBCL where multiple doses were tested (DL2: 33% vs DL1: 7%, p = 0.08). Peak serum CAR22 concentration was markedly elevated in patients who developed IEC-HS compared to those who did not (median 648 vs 42 cells/µL, p&amp;lt;0.001). Unsupervised hierarchical clustering performed on serial cytokine measurements identified a signature of IFN-γ-related cytokines including CXCL9, CXCL10, TNFα, FasL, IL6 and IL18 as being closely related in IEC-HS (Figure 1). Most cytokines in the signature peaked at D+14, consistent with onset of clinical IEC-HS. Peak levels of nearly all cytokines in the interferon-γ cluster were significantly elevated in IEC-HS patients compared to patients who did not develop IEC-HS, and this cluster contained most cytokines with significant increases in peak levels (Figure 1). Conclusions IEC-HS is a potentially life-threatening toxicity associated with CAR22 across age and underlying malignancy. The high rates of non-relapse mortality and severe infections in this study highlight the importance of understanding the immune dysregulation underlying IEC-HS. Higher CAR22 dose and expansion were associated with IEC-HS, indicating optimal dosing as a potential preventive strategy. The observed IFN-γ cytokine signature is consistent with myeloid cell activation and Th1 T-cell skewing that likely contributes to IEC-HS pathogenesis. These cytokines may be amenable to therapeutic intervention and further study of approaches targeting JAK/STAT and IFN-γ are warranted.

Ratiometric fluorescence immunoassay based on silver nanoclusters and calcein-Ce3+ for detecting ochratoxin A

Ochratoxin A (OTA), a dangerous mycotoxin, is found in many crops. It is essential to create sensitive OTA detection techniques to ensure food safety. Based on the principle of p-nitrophenol (PNP) quenched the fluorescence of bovine serum albumin silver nanocluster (BSA-AgNCs) through an internal filtering effect, and phosphate activated fluorescence of calcein-Ce3+ system, a ratiometric fluorescence immunoassay for OTA detection was developed. In this strategy, the value of F518/F640 was used as a signal for response of OTA concentration. The detection range of this strategy was 0.625–25 ng/mL, the limit of detection (LOD) was 0.04 ng/mL. This new immunoassay offered a brand-new platform for detecting OTA.

Circadian behavior and vitamin D status: a study on students at Faculty of Health, Universitas Nahdlatul Ulama Surabaya

Background: Vitamin D deficiency is still common in various community groups, including women of childbearing age. Not only because of a lifestyle that avoids the sun, but recent research has shown that circadian behavior, which includes eating and sleeping patterns, is associated with vitamin D deficiency. This study aims to provide information about female students' vitamin D status and circadian behavior to prevent further health problems. Methods: This research was quantitative, analytic, and observational, with a cross sectional study approach. The population was students of the academic year 2019 in the Faculty of Health, Universitas Nahdlatul Ulama Surabaya, in 2022. The sample size was 50 people. Data collection includes circadian behavior with parameters of sleep patterns, eating patterns, and plasma vitamin D levels. Sleep pattern data was collected using The Pittsburg Sleep Quality Index (PSQI) questionnaire. Circadian eating pattern data was taken with 24-hour recall instruments. Plasma vitamin D levels were obtained by taking blood samples analyzed using the Fluorescence Immunoassay Rapid Quantitative Test method. Data analysis was performed using the Mantel-Haenszel statistical test on the SPSS program. Results: The results showed that 80.0% of samples had a predominantly daytime (pDT) eating pattern, 64.0% had poor sleep quality, and 68.0% had vitamin D deficiency. Bivariate analysis for eating patterns and vitamin D status showed OR = 0.890 (0.197-4.012). Bivariate analysis for sleep pattern and vitamin D status showed OR value = 1.362 (0.385-4.817). However, the p-value indicates an insignificant correlation (p=0.880 and p=0.631) Conclusion: The study shows poor sleep quality and predominantly daytime eating patterns are not risk factors for vitamin D deficiency.

Accuracy of serological tests for bovine brucellosis: A systematic review and meta-analysis

The direct methods for diagnosis of bovine brucellosis have several limitations, therefore serological tests are the basis for the diagnosis of the disease. However, a meta-analysis estimating the diagnostic sensitivity (DSe) and diagnostic specificity (DSp) on the main tests used in bovine brucellosis control programs worldwide has not been performed. This systematic review and meta-analysis aimed to estimate the DSe, DSp and thereby accuracy of serological tests individually used in the diagnosis of bovine brucellosis. The databases CABI, Cochrane Library, PubMed/MEDLINE, SciELO, Scopus and Web of Science were used to select articles. The search resulted in 5308 studies, of which 71 were selected for systematic review using quality assessment tools and 65 studies were included in the meta-analysis. For the meta-analysis, 178 assays and 11 different serological tests were considered. To estimate DSe and DSp of the tests, studies were divided according to animal selection for the studies: (1) studies that carried out a random or consecutive selection of participants (noncasecontrol studies) and (2) all studies, including casecontrol studies. Considering only the non-case-control studies to estimate the DSe, the tests that exhibited the best and worst performance were the iELISA test (indirect enzyme immunoassay - bacterial suspension as antigen - BS) (96.5%, 95% CI: 94.1–97.9%) and 2ME (2- mercaptoethanol test) (85.0%, 95% CI: 79.6–89.1%), respectively; while for DSp, the FPA (fluorescence polarization assay) (99, 7%, 95% CI: 99.5–99.8%) and PCFIA tests (protein concentration fluorescence immunoassay) (78.5%, 95% CI: 70.0–85.1%) showed better and worse performance, respectively. Overall, our results showed an overestimation in the DSe and DSp of the eleven serological tests assessed when casecontrol studies were included in the meta-analysis, which is a concern considering its impacts on the time and costs associated with populational diagnosis of the diseases, since several of these tests are routinely used in the control and eradication programs of bovine brucellosis worldwide. Furthermore, the tests that exhibited the best DSe and DSp, iELISA (BS) and FPA, respectively, are relatively easy to perform and interpret and the test which showed the best overall accuracy was FPA.

Role of cardiac marker troponin I in chronic periodontitis with coronary artery disease

Link of Video Abstract: https://youtu.be/Cw5keIvNHoQ Introduction: This study aims to analyze the effect of chronic periodontitis (CP) on Troponin I levels in coronary artery disease (CAD) patients. Patients and methods: This study was a cross-sectional study. A total of 90 CAD patients at Dr. Doris Sylvanus Hospital, Palangka Raya, Central Kalimantan, Indonesia, who underwent coronary angiography and had CP, was involved in the study. CAD was examined based on significant stenosis on coronary angiography. CP was measured based on mean physical diagnostic (PD) and panoramic X-rays. The blood sample was collected to determine Troponin I level by sandwich immunodetection method with fluorescence immunoassay (FIA). Bivariate analysis of factors affecting Troponin I (with threshold ROC results) was conducted using the Chi-square test, followed by multivariate analysis. A logistic regression test was also evaluated to determine the most influential independent variable on Troponin I. Results: Based on bivariate analysis, CP and dyslipidemia significantly differed in both troponin groups. There was 53.3% mild CP at Troponin I level &lt;0.015 ng/mL, while 80% moderate-severe CP at Troponin I level &gt;0.015 ng/mL with p value=0.018. Based on multivariate analysis showed that CP had a significant effect on Troponin I. Additionally, patients with moderate-severe CP tended to have more Troponin I &gt; 0.015 ng/mL with an odds ratio (OR) of 5.528 (95% CI 1.373-22.247; p value=0.016). Conclusion: There was an effect of CP on increasing Troponin I levels in CAD patients.

NIR-II fluorescent Ag2Se polystyrene beads in a lateral flow immunoassay to detect biomarkers for breast cancer.

Fluorescent lateral flow immunoassay (LFA), one tool in point of care testing (POCT) systemsfor breast cancer, has attracted attention because it is quick, simple, and convenient. However, samples and the constituent material exhibit autofluorescence in the visible region, which is a very large obstacle in the development of fluorescent LFAs. The autofluorescence of biological samples is scarcely found in the second near-infrared (NIR-II) range and samples scatter and absorb less NIR-II light than visible light. Here, we report an NIR-II QD-LFA platform using the NIR-II fluorescent Ag2Se quantum dots (QDs) with 1020nm emission encapsulated into polystyrene beads as fluorescent probes. The NIR-II LFA platform was established to detect breast cancer tumour markers (CEA and CA153) within 15min with a low limit of detection (CEA: 0.768ngmL-1, CA153: 1.192 U mL-1), high recoveries (93.7% ~ 108.8%), and relative standard deviations (RSDs) of less than 10%. This study demonstrated the potential of NIR-II Ag2Se polystyrene beads as a fluorescent probe in LFA for rapid and accurate identification of biomarkers. They are suited for use in professional situations.

Associations between host microbiome and inflammation suggest role for host microbiome in driving COVID-19 disease severity

Systemic inflammation and innate immune activation are associated with COVID-19 disease severity. Knowledge gaps remain in the relationships between microbiome, inflammation and COVID-19 disease severity. To better characterise these associations, we performed 16SrDNA analysis of stool samples in COVID-19 subjects to explore diversity and taxanomic composition. We correlated these to host inflammatory profiles, derived from soluble plasma biomarkers measured by bead-based fluorescence and electrochemiluminescence immunoassays. Associations of microbial diversity and inflammatory biomarkers on maximal COVID-19 severity (mild, moderate v severe/critical) was explored using logistic regression and weighted gene correlation network analysis (WGCNA). Of 79 subjects, 58% were male and 88% were Caucasian with 36% experiencing mild disease, 22% moderate disease and 40% critical/severe COVID-19. Hierarchical clustering and principal component analysis (PCo) revealed distinct inflammatory clusters that were found to correlate with 4 modules of microbiome profiles. Modules 3 and 4 were associated with both older age and severe/critical disease outcomes. These modules were enriched in pathogenic and inflammatory bacteria that mapped to a pro-inflammatory biomarker cluster. In contrast, module 1 exhibited enrichment of anti-inflammatory bacteria, was associated with younger age and mild/moderate disease outcomes and mapped to a less-inflamed biomarker cluster. This study provides further insights into links between host microbiome, inflammatory responses to SARS-CoV-2 infection and clinical COVID-19 disease severity, suggesting a role for the microbiome in shaping distinct host inflammatory responses to infection.

Differences of Hepatitis B Serological Tests in Cirrhosis and Hepatocellular Carcinoma Patients

Cirrhosis and hepatocellular carcinoma (HCC) are the most common complications of chronic hepatitis B. Hepatitis B Virus (HBV) reactivation occurs in the inactive phase, characterized by reappearance of HBsAg or negative HBsAg. The prevalence of occult hepatitis B infection in cirrhosis and HCC ranges from 20% to 60%. This study aimed to analyze the differences in hepatitis B serological tests in patients with cirrhosis and HCC for diagnosis of acute or chronic hepatitis B. The current research was analytical and descriptive with a cross-sectional study design. This study involved 177 patients, including 50 cirrhosis patients and 127 HCC patients. Hepatitis B serological tests (HBsAg, HBeAg, anti-HBe, total anti-HBc) were analyzed using VIDAS instruments by the Enzyme-Linked Fluorescent Immunoassay (ELFA) method. Hepatitis B serological test results were grouped based on the interpretation of serological test results. The data were processed with the statistical test Kolmogorov-Smirnov test, independent T-test, and Chi-Square, and results with p&lt;0.05 were reported as significant. This study found that most males suffer from cirrhosis and HCC due to chronic HBV infection, with a mean age of 57. Chronic hepatitis was most common in patients with cirrhosis and HCC (71.2%). There was no significant difference in the interpretation of the hepatitis B serological test between patients with cirrhosis and HCC, with a p-value of 0.230 (p &gt; 0.05). There was no significant difference in interpreting the hepatitis B serological test between cirrhosis and HCC. Both reactive HBsAg and non-reactive HBsAg can be obtained in cirrhosis and HCC.

Circulating Inflammatory Factor Levels in the Early Phase of COVID-19 are Associated with the Progression of Respiratory Failure: A Single-Center Retrospective Study.

To evaluate the potential relationships between serum interleukin (IL)-2, IL-4, IL-6, IL-10, IL-17, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α levels and occurrence of respiratory failure in patients with early-stage COVID-19 disease. We analyzed clinical characteristics, laboratory parameters, and immunoinflammatory markers in 302 patients diagnosed with SARS-CoV-2 infection who required hospitalization at Changshu Hospital of Nantong University. IL-2, IL-4, IL-6, IL-10, IL-17, IFN-γ, and TNF-α levels in the peripheral blood of patients hospitalized five days after disease onset were measured using multiplex bead-based flow fluorescent immunoassay (MBFFI). Patients with respiratory failure had higher serum IL-4 [0 (0, 0.54) pg/mL], IL-6 [40.76 (12.33, 90.28) pg/mL], IL-10 [6.65 (4.12, 11.34) pg/mL], and IL-17 [9.48 (4.31, 12.13) pg/mL] levels than patients without respiratory failure (P=0.042, P<0.0001, P=0.012, and P=0.036, respectively). Serum IL-2, IFN-γ, and TNF-α levels were not significantly different between the two groups. The occurrence of respiratory failure was positively correlated with sex (R=0.122, P=0.034), lactic acid (R=0.193, P=0.007), white blood cell count (R=0.121, P=0.038), erythrocyte distribution width (R=0.131, P=0.024), thyrocalcitonin (R=0.280, P<0.0001), and D-dimer levels (R=0.214, P<0.0001) but negatively correlated with oxygen partial pressure (R=-0.208, P=0.004), oxygen saturation (R=-0.220, P=0.002), lymphocyte count (R=-0.129, P=0.026), and calcium (R=-0.152, P=0.042). Among the immunoinflammatory biomarkers, the occurrence of respiratory failure was positively correlated with IL-4 (R=-0.117, P=0.042), IL-6 (R=0.206, P<0.0001), IL-10 (R=0.145, P=0.012), and IL-17 (R=0.121, P=0.036) levels. Serum levels of pro-inflammatory cytokines IL-6 and IL-17 and anti-inflammatory cytokines IL-4 and IL-10 were significantly elevated in patients with respiratory failure and weakly positively correlated with the occurrence of respiratory failure. Further studies are required to explore these key immune mechanisms to help clinicians better manage acute complications, long-term sequelae, and possible future COVID-19 variants and be flexible in managing future epidemics and similar public health threats.

Controllable formation of polydopamine on carbon dots for ultrasensitive detection of alkaline phosphatase and ratiometric fluorescence immunoassay of benzocaine.

Sensing alkaline phosphatase (ALP) activity with high sensitivity and accuracy is critical for both ALP-related health and food safety supervision and the development of ALP-triggered immunoassay platforms. Herein, an ultrasensitive ratiometric fluorescence (RF) sensing system based on the controllable formation of luminescent polydopamine and efficient quenching of carbon dots was proposed for the ALP activity assay, achieving quantitative detection in the range of 0.01-100mU/L. Furthermore, this RF sensing system was integrated with an ALP-based ELISA platform to construct an RF-ELISA for benzocaine, a potentially abused anesthetic in edible fish, and ultrasensitive assay at the level of fg/mL was realized. This ratiometric strategy-based platform effectively shields various interferences through the self-calibration effect, thus providing more accurate and reliable quantification results. This study not only offers an efficient method for ultratrace detection of ALP and benzocaine but also proposes a universal platform for ultrasensitive detection of diverse targets in food analysis by replacing the recognition unit.

Inner filter effect-based fluorescence immunoassay using nanobody-alkaline phosphatase fusion and gold nanoclusters for detecting ochratoxin A in pepper

Inner filter effect-based fluorescence immunoassay using nanobody-alkaline phosphatase fusion and gold nanoclusters for detecting ochratoxin A in pepper

Construction of L-type lectin displaying Saccharomyces cerevisiae for Vibrio parahaemolyticus agglutination.

The utilization of Aga1P anchor protein in the display system for expressing heterologous proteins on the surface of Saccharomyces cerevisiae has been shown to be an ideal approach. This system has the ability to improve the expression of target proteins beyond the cell surface, resulting in increased activity and stability of the expression system. Recent studies have demonstrated that a new L-type lectin from Litopenaeus vannamei (LvLTLC1) has been found to possess the capability of agglutinating Vibrio parahaemolyticus, a pathogen responsible for causing acute hepatopancreatic necrosis disease (AHPND) in shrimp. In this study, LvLTLC1 protein was designed to be expressed on the surface of S. cerevisiae via Aga1P anchor. The expression of LvLTLC1 protein on the surface of S. cerevisiae::pYIP-LvLTLC1-Aga1P was confirmed through the use of analytical techniques including SDS-PAGE, dot blot, and fluorescent immunoassay with LvLTC1-specific antibody. Subsequently, the newly generated yeast strain was evaluated for its ability to agglutinate V. parahaemolyticus and A. hydrophila. The obtained results indicated that S. cerevisiae expressing LvLTLC1 protein on its surface had the ability to agglutinate both AHPND-causing V. parahaemolyticus and A. hydrophila. This newly generated yeast strain could be served as a feed supplement for controlling bacteria in general and AHPND in particular.