Immunoassay Method Research Articles

Diagnostic Accuracy of IgA Anti-Transglutaminase Assessed by Chemiluminescence: A Systematic Review and Meta-Analysis.

A new chemiluminescence immunoassay method (CLIA) for detecting IgA anti-transglutaminase (atTG IgA) in celiac disease (CD) has prompted inquiries into its diagnostic performance. We conducted a systematic review and meta-analysis comparing CLIA with traditional enzyme-linked immunosorbent assay (ELISA) and fluorescence enzyme immunoassay (FEIA). We searched PubMed, Medline, and Embase databases up to March 2024. The diagnostic references were intestinal biopsy and ESPGHAN guidelines. We calculated the sensitivity and specificity of atTG IgA assessed by CLIA and the odds ratio (OR) between the assays. Eleven articles were eligible for the systematic review and seven for the meta-analysis. Sensitivity and specificity of atTG IgA CLIA-assay were 0.98 (95% CI, 0.95-0.99) and 0.97 (95% CI, 0.94-0.99), respectively. The sensitivity of atTG IgA antibody detection did not significantly vary across the three assay modalities examined (CLIA vs. ELISA OR: 1.08 (95% CI, 0.56-2.11; p = 0.8); CLIA vs. FEIA OR: 6.97 (95% CI, 0.60-81.03; p = 0.1). The specificity of atTG IgA assessed by FEIA was higher than for CLIA (OR 0.17 (95% CI, 0.05-0.62); p < 0.007). According to the systematic review, normalization of atTG IgA levels in CD patients following a gluten-free diet was delayed when using CLIA compared to ELISA and FEIA methods. Conflicting findings were reported on the antibody threshold to use in order to avoid biopsy confirmation.

The Correlation Between Interleukin-6 (IL-6) Levels and the Extent of Coronary Lesions Based on SYNTAX Scores in Patients with Acute Coronary Syndrome (ACS)

&lt;p&gt;&lt;strong&gt;Introduction: &lt;/strong&gt;Atherosclerosis plays an important role in the pathogenesis of acute coronary syndrome (ACS). Inflammation plays a causal role in atherosclerosis. There are many inflammatory markers related to ACS, including C-reactive protein (CRP), homocysteine, matrix metal loproteinases, interleukin (IL)-6, IL-8, and IL-27. IL-6 is the sign of synthesis of acute phase proteins. Recently, the clinical trials related to ACS showed that IL-6 may be an indicator of increased risk for ACS. An Interleukin-6 (IL-6) examination is can be used to assess risk factors for acute coronary syndrome (ACS). Interleukin-6 is a marker of local inflammation of coronary plaque and peripheral blood circulation which increases the occurrence of coronary heart disease causing plaque instability and rupture. The SYNTAX scoring system has correlation assessed through angiography assessing the complexity of the severity of coronary artery stenosis. This study aims to determine the between Interleukin-6 (IL-6) levels and the extent of coronary lesions based on the SYNTAX score in patients with acute coronary syndrome (ACS).&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;This is a cross-sectional study of 45 patients with acute coronary syndrome (ACS) who were treated at H. Adam Malik General Hospital, Medan (February 2023 – May 2023). IL-6 levels were examined using the Electrochemiluminescent Immunoassay (ECLIA) method using Cobas e 411. The extent of the coronary lesion was calculated based on the SYNTAX score from coronary angiography which was calculated by two invasive cardiologist as suggested before using an online calculator. We used the Spearman correlation test and Mann Whitney test to analyze the data.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;The were 39 male subjects (86.7%) total.&lt;strong&gt; &lt;/strong&gt; The mean age of the subjects was 54.89 ± 10.3 years old. Most of the subjects experienced STEMI in total 28 people (62.2%). The Median IL-6 levels was 19.22 pg/mL while the median of SYNTAX score level was 13.00. By using &lt;em&gt;cross-sectional&lt;/em&gt; study and the spearman correlationa test, there is a significant correlation between IL-6 and the SYNTAX score with the correlation value obtained r = 0.343 (p = 0.021).&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;There is a significant relationship between IL-6 and the extent of coronary lesions based on the SYNTAX score in patients with acute coronary syndrome.&lt;/p&gt;&lt;p&gt; &lt;/p&gt;

Screening and identification of peptidyl arginine deiminase 4 inhibitors from herbal plants extracts and purified natural products by a trypsin assisted sensitive immunoassay based on streptavidin magnetic beads

Peptidyl arginine deiminase 4 (PAD4) plays a critical role in many autoimmune diseases including rheumatoid arthritis. Herein, a trypsin assisted highly immunoassay method was established to determine PAD4 activity and screen potent inhibitors from herbal plants extracts and purified natural products. The method was applied to determine endogenous PAD4 activity in both cell and tissue lysates, as well as the inhibitory effects of 20 herbal plants and 50 purified natural products. The Cinnamomi ramulus extract showed strongest inhibitory potency with IC50 value lower than 5 μg/mL. Meanwhile, pyrroloquinoline quinone (PQQ), widely used as a dietary supplement, was discovered as a promising PAD4 inhibitor with an IC50 value lower than 4 μM. The inhibition kinetic analysis, drug affinity response target stability (DARTS) and molecular docking were performed to confirm the interaction between PQQ and PAD4. This method has great potential for researchers to monitor activities and discover potential inhibitors of PAD4.

A miniaturized platform based on reciprocating-flowing immunobinding for ultrafast, highly-sensitive and on-site immunoassay

Immunoassays are considered important methods for detecting disease biomarkers for biomedical diagnosis. Particularly, enzyme-linked immunosorbent assay (ELISA) is a commonly-used immunoassay method due to its high reliability and low cost. Nowadays, automatic analyzers integrating all steps of ELISA can achieve detection without manual operation to save labor, increase detection efficiency and avoid manual error. However, most ELISA analyzers require more than 30 min for one detection, which are not fast enough for on-site point-of-care testing (POCT). Herein, based on the reciprocating-flowing immunobinding strategy, we developed a miniaturized reciprocating-flowing POCT platform (RF-POCT platform) for rapid immunodetection. RF-POCT platform, which included an electronic control module, a fluid control module, a mechanical control module and a detection module, enabled fast immunobinding of antigen and antibody. Taking ELISA as an example of immunoassay methods for RF-POCT platform, we developed a miniaturized reciprocating-flowing ELISA analyzer (RF-ELISA analyzer) to automatically detect SARS-CoV-2 nucleocapsid protein (SARS-CoV-2 N protein) and dust mite-specific immunoglobulin E (DM-IgE), with the LOD of 5.2–6.6 pg/mL and the CV less than 3 %. The detection required only 3.5 min. Afterwards, we detected the SARS-CoV-2 N protein in nasal swab samples and the DM-IgE in serum samples; the specificities were 85.7–100 %, the sensitivities were 96.3–100 %. The quantitative results from RF-ELISA analyzer were generally-consistent with those from large ELISA analyzers in hospitals. Therefore, RF-ELISA analyzer demonstrated excellent performance on ultrafast, highly-sensitive, and on-site immunoassay, with potential to support on-site POCT for primary hospitals and community clinics.

Hybrid Mn Atomic Clusters/Single-Dispersed Atoms with Dual Antioxidant Activities for a Chemiluminescent Immunoassay.

Single-dispersed atoms (SDAs) as catalysts have drawn extensive attention due to their ultimate atom utilization efficiency and desirable catalytic capability. Atomic clusters (ACs) with potential multiple enzyme-like activities also display great practicability in catalysis-based biosensing. In this work, hybrid Mn ACs/SDAs were implanted in the frameworks of defect-engineered MIL 101(Cr) modulated by excess acetic acid, with a high loading capability of 13.9 wt %. Distinctively, Mn SDAs display weak superoxide dismutase (SOD)-like activity for specifically eliminating superoxide anion (O2•-), while Mn ACs/SDAs display both catalase-like and SOD-like activities for remarkable elimination of total reactive oxygen species (ROS) due to the cooperative effect of the two atom-scale catalytic sites. Thus, Mn ACs/SDAs can efficiently inhibit the chemiluminescent (CL) emission of multiple ROS-mediated luminol systems with a superior quenching rate of 85.5%. To validate the practicability of Mn ACs/SDAs for a sensitive CL assay, an immunoassay method was established to detect acetamiprid by using Mn ACs/SDAs as signal quenchers, which displayed a quantification range of 10 pg mL-1-25 ng mL-1 and a detection limit of 3.3 pg mL-1. This study paves an avenue for developing ACs/SDAs with multiple antioxidant activities that are suitable for application in biosensing.

The Influence of Growth Hormone and Insulin-like Growth Factor 1 on Anthropometric Parameters in Children with Type 1 Diabetes Mellitus

The goal is to study the indicators of somatotropic hormone and insulin-like growth factor-1 to assess the effect on anthropometric indicators of children with type 1 diabetes mellitus. 74 children with type 1 diabetes mellitus and 14 children in the control group were examined. Average age 12±3.5 years. The assessment of physical development was carried out on the basis of an assessment of anthropometric indicators — height and body weight with the calculation of BMI and compared with centile tables. Determination of the concentration of hormones such as GH and IGF-1 was determined by special ELISA kits using the enzyme immunoassay method using an analyzer (Mindray MR-96A). The obtained data were subjected to statistical processing. It was revealed that 8 (36.4%) boys and the majority of girls 14 (63.6%) achieved the target level of glycated hemoglobin. Children with T1DM are statistically significant and stunted in height and weight compared to healthy children in the control group.

Increased serum ferritin is associated with severity of orbital disease in COVID-19-associated rhino-orbito-cerebral mucormycosis: A quantitative analysis.

Effect of serum ferritin on severity of coronavirus disease 2019 (COVID-19)-associated rhino-orbito-cerebral mucormycosis. To study the association between increased serum ferritin and severity of orbital disease in COVID-19-associated rhino-orbito-cerebral mucormycosis. A cross-sectional study. Hundred (n) out of 155 treatment-naive patients of COVID-19 infection presenting with the signs and symptoms of rhino-orbito-cerebral mucormycosis were enrolled in study. Based on the classification proposed by Honavar, the study patients were classified into four stages: Stage 1: involvement of the nasal mucosa (n = 11), Stage 2: involvement of paranasal sinuses (n = 14), Stage 3: involvement of the orbit (n = 37), Stage 4: involvement of the central nervous system (n = 38). Stage 3 was further divided into four substages: 3a: nasolacrimal duct, medial orbit, vision unaffected (n = 4); 3b: diffuse orbital involvement (>1 quadrant or >2 structures), vision unaffected (n = 15); 3c: central retinal artery occlusion or ophthalmic artery occlusion, superior ophthalmic vein thrombosis, involvement of superior orbital fissure, inferior orbital fissure, orbital apex, diminution or loss of vision (n = 13); 3d: bilateral orbital involvement (n = 5). Fasting blood sugar (FBS), postprandial blood sugar (PPBS), and inflammatory markers (serum ferritin, interleukin-6, C-reactive protein, and D-dimer) were assessed. Serum level of ferritin was analyzed by using chemiluminescence immunoassay method. Mean FBS (mg/dl) was 165.03 ± 70.43 for stage 1, 185.67 ± 64.82 for stage 2, 159.05 ± 68.60 for stage 3, and 158.20 ± 62.05 for stage 4. Mean PPBS (mg/dl) was 238.70 ± 141.29 for stage 1, 252 ± 103.69 for stage 2, 257.09 ± 103.48 for stage 3, and 229.53 ± 76.81 for stage 4. Mean serum ferritin (µg/l) was 302.67 ± 266.95 in stage 1, 444.19 ± 116.36 in stage 2, 504.85 ± 205.99 in stage 3, and 825.95 ± 777.30 in stage 4. A statistically significant increase in serum ferritin levels with severity of disease (P = 0.005) was noted. Similar trend was observed in substages of stage 3. Pearson correlation analysis showed a positive correlation between serum ferritin and severity of disease (P = 0.0007). Increased serum ferritin was significantly independently associated with severity of orbital disease in COVID-19-associated rhino-orbito-cerebral mucormycosis.

Drugs in blood and urine samples from victims of suspected exposure to drink spiking: A prospective observational study from Oslo, Norway.

People regularly contact emergency medicine services concerned that they have been exposed to drink spiking, i.e., exposure to drugs without their knowledge or permission. We identified drugs in blood and urine samples from patients suspecting exposure to drink spiking, with special consideration for drugs not reported taken by the patient (unreported drugs). From September 2018 to May 2019, we collected blood and urine samples from patients 16 years or older presenting at an emergency clinic in Oslo, Norway, within 48 hours of suspected exposure to drink spiking. We also collected information on ethanol ingestion and drugs taken. Blood samples were analyzed for 20 classical recreational drugs using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and an automated enzymatic method for ethanol. Urine samples were analyzed using immunoassay methods and a specific gas chromatography mass spectrometry (GCMS) method for gammahydroxybutyrate (GHB). From 100 included patients (median age 24 years, 62 females), we collected 100 blood samples and 72 urine samples. Median time since exposure was 5 hours. Unreported drugs were found in 15 patients. Unreported drugs in the blood samples were clonazepam in 3, methylenedioxymethamphetamine (MDMA) in 3, amphetamine in 2, tetrahydrocannabinol (THC) in 2, tramadol in 1, cocaine in 1, and methamphetamine in 1. Unreported drugs in the urine samples were cocaine in 5, amphetamine in 4, ecstasy in 3, and cannabis in 2. Ethanol was found in 69 patients, all reporting ethanol ingestion. Median blood ethanol concentration was higher in patients with no unreported drugs detected, 1.00‰ (interquartile range (IQR) 0-1.52) vs. 0‰ (IQR 0-0.46) (p<0.001). GHB was not detected. Unreported drugs, possibly used for drink spiking, were found in 15% of patients. Blood ethanol concentration was higher when no unreported drugs were found. GHB was not detected in any patient.

Glyphosate contamination: implications for honeybee Apis mellifera and consumers in Southeastern Mexico

In a study conducted from June 2021 to May 2022 in two apiaries in southeastern Mexico, levels of glyphosate residues in pollen collected by bee Apis mellifera were analyzed to assess potential risks to both bees and humans. The analysis used an immunoassay method after residue extraction using the QUECHERS method. The results revealed the presence of glyphosate in all samples, with concentrations ranging between 3.71 and 7.29 μg/kg. However, risk analysis, as indicated by the pollen hazard quotient, suggested that these quantities did not pose a serious threat to bees or humans. The levels were within acceptable limits of the acceptable daily intake (ADI), acute reference dose (ARfD) and acceptable operator exposure level (AOEL). In conclusion, although this study did not find any significant association between glyphosate and potential risks for both humans and bees, its persistence in the environment is demonstrated. Glyphosate levels at the study site were low, suggesting minimal risk to both species. However, the wide distribution of glyphosato in the region makes it necessary to emphasize long-term studies to understand the possible chronic effects of the pesticide on all species in the area.

High prevalence of hepatitis B and HIV among women survivors of sexual violence in South Kivu province, eastern Democratic Republic of Congo.

Limited data are available on the prevalence rates of hepatitis B and acquired immunodeficiency syndrome (AIDS) among women survivors of sexual violence (WSSV) in South Kivu province, in the eastern part of the Democratic Republic of Congo (DRC), where armed conflicts persist. Here, we aimed to assess the prevalence of these two infections in this vulnerable local population. A total of 1002 WSSV, aged from 18 to 70 years old were enrolled from May 2018 to May 2020 at three healthcare facilities (Panzi, Mulamba and Bulenga hospitals), which are called "The One-Stop Centre Care Model" for the management of sexual violence in South Kivu. Blood samples were collected and tested for hepatitis B virus (HBV) and human immunodeficiency virus (HIV) antigens and antibodies using enzyme-linked immunoassay (ELISA) methods. Subsequently, viral load quantification for HBV and HIV were performed using the GeneXpert. Univariate and multivariate logistic regression models were used to assess factors associated with HIV-positive and HBV-positive status. For HBV, overall prevalence was 8.9% (95% CI; 7.2-10.8%), 32.1% (95% CI; 29.3-35.0%), and 14.5% (95% CI; 12.3-16.8%) for HBsAg, anti-HBc and anti-HBs antibodies, respectively. Among the 89 HBsAg-positive patients, 17 (19.1%) were HBeAg-positive. The median age of individuals with a positive HBsAg test was higher than those with a negative test (median: 40 years (IQR 30-52) compared to 36 years (IQR 24-48)). Risk factors for HBV infection were age (≥35 years) (AOR = 1.83 [1.02-3.32]; p = 0.041), having no schooling (AOR = 4.14 [1.35-12.62]; p = 0.012) or only primary school-level (AOR = 4.88 [1.61-14.75]; p = 0.005), and multiple aggressors (AOR = 1.76 [1.09-2.84], p = 0.019). The prevalence of HIV was 4.3% [95% CI: 3.1-5.7%]. HIV/HBV co-infection occurred only in 5 individuals (0.5%). The HBV viral load was detectable (> 1 log10 UI/mL) in 61.8% of HBsAg-positive subjects and 64.8% HIV-positive subjects had a high viral load (≥ 3 log10 copies/mL). This study revealed a high prevalence of HBV and HIV infections among WSSV in South Kivu. The results generated highlight the urgent need for systematic screening of HBV and HIV by integrating fourth-generation ELISA tests in HIV and HBV control programs.

Methods of Analysis of Phytoestrogenic Compounds: An Up-to-Date of the Present State

Phytoestrogens, natural compounds with structural similarity to 17-β-estradiol, are known to have potential health benefits, including in hormone-dependent malignancies. The therapeutic interest and some safety concerns observed triggered the need to develop accurate methods to assess their level in different matrices. This narrative review examines the existing analytical methods used to isolate, quantify, and characterize phytoestrogens and their metabolites in plants, foods, and biological samples. Different conventional and modern extraction techniques, such as ultrasonication-assisted extraction, supercritical fluid extraction, or enzyme-assisted extraction, were presented and compared. The advantages and limitations of the existing analytical methods, such as high-performance liquid chromatography using different sensitive detectors, gas chromatography often coupled with mass spectrometry, and immunoassay methods, are highlighted, along with the need for ongoing research to improve the sensitivity and selectivity of the analysis.

Exploring the Association between Depression, Suicidality, and Serum Brain-Derived Neurotrophic Factor, FAM19A5, Total Antioxidant Status, Total Oxidant Status, Oxidative Stress Index, and Cytokine Levels: A Case-Control Study of Drug-Naive Adolescents with First-Episode Major Depressive Disorder

Introduction: Considering the importance of neuroinflammation and neurodegeneration in the pathophysiology of major depressive disorder (MDD), peripheral blood biomarkers are promising for the prediction of diagnosis and treatment outcomes. We aimed to elucidate the neuroinflammatory pathophysiology of depression by evaluating serum levels of FAM19A5 as a new biomarker of inflammatory activation, proinflammatory cytokines, brain-derived neurotrophic factor (BDNF), and oxidative stress parameters. Methods: Adolescents diagnosed with first-episode drug-naive MDD (n = 35) were compared neurobiologically healthy control group (n = 33). Serum FAM19A5 levels, cytokine levels, BDNF and oxidative stress parameters were evaluated using the enzyme-linked immunoassay method. All participants were assessed with the Level-2 Depression Severity Scale, Sleep Disturbance Scale, Somatic Symptom Scale. Results: BDNF levels were significantly higher in the patient group compared to the control group. While BDNF showed a positive correlation with all scale scores; BDNF was significantly higher in the suicide risk groups than the control group. IL-1β levels displayed a negative correlation with the severity of sleep disturbances. Conclusions: In adolescents with MDD, inflammatory and oxidative stress markers were not raised in peripheral blood, unlike in adults. However, BDNF levels, which typically decrease in neurodegenerative conditions, were higher in those with MDD.

Serum DNase1, sTNFR1 and sTNFR2 as Risk Factors for Lupus Nephritis in Systemic Lupus Erythematosus Patients

BACKGROUND: Early detection and management of lupus nephritis (LN) in systemic lupus erythematosus (SLE) are essential to prevent irreversible kidney damage and improve patient outcomes; therefore, identifying reliable biomarkers to predict LN is paramount. However, there are still relatively few studies examining the potential biomarkers for LN in SLE patients. This study was conducted to investigate serum deoxyribonuclease I (DNase), soluble tumor necrosing factor 1 (sTNFR1) and soluble tumor necrosing factor 2 (sTNFR2) as a risk factor for LN in SLE patients.METHODS: A case-control study involving SLE patients aged 20-60 years was conducted. Blood was withdrawn from each subject for the measurement of serum level of DNase1, sTNFR1, and sTNFR2 that was performed using enzyme-linked immunoassay (ELISA) methods. Data was then analyzed using Chi-Square test and logistic regression tests.RESULTS: A total of 22 patients with LN and 22 without LN were included. The cut-off value for DNase1, sTNFR1, and sTNFR2 were 5.05 ng/mL, 6.52 ng/mL, and 7.02 ng/mL, respectively. The risk factors of LN in SLE patients were the low level of serum DNase1 (aOR=6.64; 95%CI: 1.25-35.29; p=0.026), low level of serum sTNFR1 (aOR=8.12; 95% CI: 1.56-42.10; p=0.013), and low level of serum sTNFR2 (aOR=5.57; 95%CI: 1.03-30.11; p=0.046).CONCLUSION: Serum DNase1 lower than 5.05 ng/mL, sTNFR1 lower than 6.52 ng/mL, and sTNFR2 lower than 7.02 ng/mL were risk factors for lupus nephritis in SLE patients. Hence, serum DNase1, sTNFR1 and sTNFR2 could be used as risk factors predictors for LN in SLE patients.KEYWORDS: DNase1, sTNFR1, sTNFR2, SLE, lupus nephritis

Association of Age at Menarche with Inflammation and Glucose Metabolism Biomarkers in U.S. Adult Women: NHANES 1999-2018.

Early age at menarche (AAM) is a risk factor for type 2 diabetes later in life, but the pathogenic pathways that confer increased risk remain unknown. We examined the associations between AAM and inflammatory and glucose metabolism biomarkers among U.S. adult women who were free of diabetes. Using the National Health and Nutrition Examination Survey (NHANES) 1999-2018, 19,228 women over 20 years old who were free of self-reported cancer and diabetes were included in this cross-sectional analysis. AAM was the self-reported age at first menstruation. CRP, fasting glucose, fasting insulin, and ferritin levels were measured as biomarkers of inflammation and glucose metabolism in adult blood samples using latex-enhanced nephelometry, enzymatic, and immunoassay methods. Multiple linear regression was used to relate AAM to the biomarkers. The median age at the time of blood sample collection was 44 years (IQR, 33-62). After age adjustment, there was an association between a lower AAM and higher CRP (P-trend=0.006); fasting glucose (P-trend<0.0001); fasting insulin (P-trend <0.0001); and ferritin (p-trend<0.0001). These remained significant after additional adjustment for demographic, reproductive, lifestyle, and adiposity variables, except for ferritin. Smoking modified the effect of AAM on CRP (p-interaction = 0.014), fasting insulin (p-interaction <0.001), and fasting glucose (p-interaction<0.001). In stratified analysis, the observed associations became more pronounced in non-smokers, while they were attenuated to non-significance in active smokers. Earlier age at menarche is associated with an unfavorable inflammatory and glucose metabolic biomarker profile in a nationally representative sample of adult women free of diabetes, especially among non-smokers.

Epizootic situation and molecular genetic characteristics of the rabies virus isolated from the brain of domestic reindeer in Yakutia

Rabies was registered among horses, domestic reindeer and dogs on the territory of the Republic of Sakha (Yakutia) in 2001– 2021. Rabies virus (Rabies lyssavirus) was detected in biomaterial from wild animals in Arctic foxes, red foxes, wolves, colts, ermine and sables. In the Arctic zone of Yakutia in 2023, rabies virus was isolated from a brain sample from one of the domestic reindeer of a herd showing clinical signs of severe agitation and aggression to humans. It is assumed that the reindeer was infected with rabies virus when bitten by a rabies-sick Arctic fox, as the range of distribution of these animals in the region is quite large. Rabies virus detection studies were conducted using polymerase chain reaction, enzyme immunoassay and fluorescent antibody method. The diagnosis of rabies was confirmed in all diagnostic tests. The nucleic acid fragments were isolated from the positive samples by the PCR method and their nucleotide sequences were determined. Analysis of the nucleotide sequences of the viral genome revealed phylogenetic relatedness of the isolated rabies virus isolate in the Arctic zone of Yakutia with the rabies viruses circulating in some areas of the North American continent, including Greenland (two isolates), Canada (six), and the United States (three). The isolate of the rabies virus from the brain of domestic reindeer in the Arctic zone of Yakutia belongs to the group of strains of American origin (USA1) and is phylogenetically close to the strains isolated in Greenland, Canada, and the USA. The studied virus isolate territorially corresponds to the known antigenic variant 3, established earlier in the north of the Asian part of Russia, in particular in Yakutia.

Non-Surgical Periodontal Therapy's Influence on Alpha-Synuclein and Inflammatory Marker Levels: A Pilot Study.

Background: Microbial dysbiosis may contribute to alpha-synuclein (α-Syn) homeostasis disruption, yet the burden of inflammatory periodontal infection and its treatment have never been studied in this regard. We aimed to compare the cytokine and α-Syn levels in the saliva and blood of patients with periodontitis who underwent non-surgical periodontal therapy (NSPT) and those of their healthy counterparts. Methods: Periodontal examination and saliva and blood sample collection were carried out in incoming patients at a university clinic. The periodontitis group (PG) received NSPT. The sample collection and periodontal observation were repeated 30 days after. IL-6, IL1-β and total α-Syn were quantified using immunoassay methods. The periodontal inflamed surface area (PISA) was calculated as a proxy for periodontal inflammation. Results: Eleven participants formed the PG, and there were fifteen healthy controls (HC). At baseline, no correlation between salivary and plasma α-Syn was found. The salivary α-Syn levels revealed a tendency to decrease 30 days after, particularly in the PD cases. The variation in PISA and α-Syn showed significant correlation. Salivary α-Syn correlated negatively with salivary IL-6 levels at both timepoints in the total sample (rho = -0.394 and rho = -0.451) and in the HC (rho = -0.632 and rho = -0.561). Variations in plasma IL-6 and α-Syn were negatively correlated (rho = -0.518) in the healthy participants. Baseline plasma IL1-β negatively correlated with plasmatic α-Syn at 30 days in the HC (rho = -0.581). Conclusions: Salivary and plasma α-Syn bioavailability operate independently, and periodontal diagnosis was not a confounding factor. Salivary α-Syn levels were significantly affected by NSPT, contrary to plasma levels. These results should be confirmed in future larger and prospective studies.

Atomically Co-dispersed nitrogen-doped carbon for sensitive electrochemical immunoassay of breast cancer biomarker CA15-3.

The development of an ultrasensitive and precise measurement of a breast cancer biomarker (cancer antigen 15-3; CA15-3) in complex human serum is essential for the early diagnosis of cancer in groups of healthy populations and the treatment of patients. However, currently available testing technologies suffer from insufficient sensitivity toward CA15-3, which severely limits early large-scale screening of breast cancer patients. We report a versatile electrochemical immunoassay method based on atomically cobalt-dispersed nitrogen-doped carbon (Co-NC)-modified disposable screen-printed carbon electrode (SPCE) with alkaline phosphatase (ALP) and its metabolite, ascorbic acid 2-phosphate (AAP), as the electrochemical labeling and redox signaling unit for sensitive detection of low-abundance CA15-3. During electrochemical detection by differential pulse voltammetry (DPV), it was found that the Co-NC-SPCE electrode did not have a current signal response to the AAP substrate; however, it had an extremely favorable response current to ascorbic acid (AA). Based on the above principle, the target CA15-3-triggered immunoassay enriched ALP-catalyzed AAP produces a large amount of AA, resulting in a significant change in the system current signal, thereby realizing the highly sensitive detection of CA15-3. Under the optimal AAP substrate concentration and ALP catalysis time, the Co-NC-SPCE-based electrochemical immunoassay demonstrated a good DPV current for CA15-3 in the assay interval of 1.0 mU/mL to 10,000 mU/mL, with a calculated limit of detection of 0.38 mU/mL. Since Co-NC-SPCE has an excellent DPV current response to AA and employs split-type scheme, the constructed electrochemical immunoassay has the merits of high preciseness and anti-interference, and its clinical diagnostic results are comparable to those of commercial kits.

Clinical Performance of the Line Immunoassay and Digital Liquid Chip Method for Detecting Autoimmune Liver Disease Autoantibodies.

The identification of autoantibodies associated with autoimmune liver disease (ALD) is crucial for diagnosis and management. Various laboratory methods have been introduced to detect autoantibody profiles. However, the variable performance of these assays may create challenges for clinicians and patients. To investigate the concordance rates and diagnostic performance of 2 commercially available assays, line immunoassay (LIA) and digital liquid chip method (DLCM), in patients with ALD. A total of 291 serum samples were collected, consisting of 180 sera from patients with ALD and 111 sera from controls. The samples were detected through LIA and DLCM. The agreement and diagnostic performance of each assay were analyzed. There was substantial to almost perfect agreement among prevalent autoantibodies (anti-mitochondrial antibody M2, antibodies against gp210, Sp100, and Ro52). Nevertheless, the Cohen κ coefficient of some uncommon autoantibodies (anti-LKM-1, anti-LC-1, and anti-SLA/LP) between the 2 methods was not ideal. LIA showed slightly better sensitivity, accuracy, and negative predictive value, while DLCM exhibited slightly higher specificity and positive predictive value. LIA and DLCM demonstrated comparable performance for the detection of common ALD-related autoantibodies. LIA seemed to be more sensitive, while DLCM displayed more specificity. However, standardization of ALD autoantibody detection still faces challenges between these diverse detection systems. Comprehensive interlaboratory validation is essential to mitigate potential misunderstanding and confusion among patients and clinicians.

The prooxidant-antioxidant balance in diagnosis and developmental prognosis of premature neonates with asphyxia.

The antioxidant system in a preterm neonate is premature. The imbalance between the prooxidant and antioxidant systems can make these neonates prone to oxidative stress. Birth asphyxia is one of the factors that can disturb this balance. We studied the prooxidant-antioxidant balance (PAB) in the diagnosis and developmental prognosis of preterm neonates with asphyxia. This cohort study has been conducted between 2016 and 2022 with 2 years follow-up on 183 premature neonates admitted to Ghaem Hospital Mashhad, by using a convenience sampling method. The data-collection tool and the researcher-made checklist included the mothers' and the neonate's information, and the third segment included laboratory information. PAB was studied by using standard solutions and the Enzyme immunoassays (ELISA) method. After discharging the newborns from the hospital, they were under follow-up at 6 months, 12 months, 18 months, and 24 months, by using the Denver II test. PAB was compared among newborns with asphyxia, those without asphyxia, and also newborns with normal and abnormal outcomes in both groups. The mean ± standard deviation of the PAB factor reported is as follows: in newborns without asphyxia (21.00 ± 18.14 HK), those with asphyxia (31.00 ± 45.42 HK), in newborns with asphyxia having abnormal outcomes (40.00 ± 60.84 HK), and those having normal outcomes (21.00 ± 18.67 HK) (P ≤ 0.05). PAB results >25 HK have been used for the diagnosis of asphyxia prognosis in newborns, with 83.3% sensitivity and 81% specificity. The PAB index showed a significant increase after asphyxia. It can be used as a diagnostic marker for the prognosis of premature newborns with asphyxia. Thus, diagnosis and prognosis of asphyxia in premature newborns can be predicted by using the PAB index.

All-in-one platform: Versatile, Easy, and User-friendly System (VEUS) based on automated and expert-independent antibody immobilization and immunoassay by utilizing customized movement of magnetic particles.

The ELISA is the most worldwide method for immunoassay. However, the ELISA is losing ground due to low reproducibility of manual experimental processes in both R&D and IVD areas. An automated platform is a good solution, but there are still limitations owning to extremely high cost and requiring large space to set up especially for a small size laboratory. Here, we present a novel all-in-one platform called "VEUS" settable on the laboratory table that offers comprehensive automation of the entire multiplex immunoassay process by exploiting antibody conjugated magnetic particles, quality control and then immunoanalytical reaction, thereby enhancing detection sensitivity and high reproducibility. As a proof of concept, the system exhibits a sensitive LOD of 0.6 and 3.1 pg mL-1 within 1 h run, comparable precision that of molecular diagnostic systems based on PCR method, enabling rapid multiplex diagnosis of Influenza A, Influenza B, and COVID-19 viruses with similar symptoms. Through automation by the all-in-one system, it can be used by novice users, something innovative for immunoassays, relying heavily on user experience. Furthermore, it can contribute to streamline entire immunoassay processes of diverse biomarkers with high reproducibility and convenience in laboratories.