High Performance of SARS-Cov-2N Protein Antigen Chemiluminescence Immunoassay as Frontline Testing for Acute Phase COVID-19 Diagnosis: A Retrospective Cohort Study

Objectives: COVID-19 emerged and rapidly spread throughout the world. Testing strategies focussing on patients with COVID-19 require assays that are high-throughput, low-risk of infection, and with small sample volumes. Antigen surveillance can be used to identify exposure to pathogens and measure acute infections.Methods: A total of 914 serum samples, collected from 309 currently infected COVID-19 patients, 48 recovered ones, and 410 non-COVID-19 patients, were used to measure N protein antigen levels by a chemilumineseent immunoassay. Diagnostic performances were analyzed in different periods after onset.Results: There was a high level of N protein antigen in COVID-19 patients (0.56 COI), comparing to the recovered patients (0.12 COI) and controls (0.19 COI). In receiver-operating characteristic curve analysis, the area under the curve of serum N protein antigen was 0.911 in the first week after onset. In this period, Sensitivity and specificity of serologic N protein antigen testing was 76.27 and 98.78%. Diagnosis performance of specific antibodies became better from the third week after onset. Subgroup analysis suggested that severe patients had higher levels of antigens than mild patients.Conclusions: High level of serum antigen suggested early infection and serious illness. Serum N protein antigen testing by chemiluminescence immunoassay is considered as a viable assay used to improve diagnostic sensitivity for current patients.

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Sphingolipid Analysis Indicate Lactosylceramide as a Potential Biomarker of Inflammatory Bowel Disease in Children

Biomolecules ◽

10.3390/biom10071083 ◽

2020 ◽

Vol 10 (7) ◽

pp. 1083

Author(s):

Aleksandra Filimoniuk ◽

Agnieszka Blachnio-Zabielska ◽

Monika Imierska ◽

Dariusz Marek Lebensztejn ◽

Urszula Daniluk

Keyword(s):

Inflammatory Bowel Disease ◽

Bowel Disease ◽

High Performance ◽

Area Under The Curve ◽

Study Groups ◽

Diagnostic Value ◽

Serum Samples ◽

Specificity And Sensitivity ◽

Potential Biomarker ◽

Inflammatory Bowel

An altered ceramide composition in patients with inflammatory bowel disease (IBD) has been reported recently. The aim of this study was to evaluate the concentrations of sphingolipids in the serum of treatment-naive children with newly diagnosed IBD and to determine the diagnostic value of the tested lipids in pediatric IBD. The concentrations of sphingolipids in serum samples were evaluated using a quantitative method, an ultra-high-performance liquid chromatography-tandem mass spectrometry in children with Crohn’s disease (CD) (n=34), ulcerative colitis (UC) (n = 39), and controls (Ctr) (n = 24). Among the study groups, the most significant differences in concentrations were noted for C16:0-LacCer, especially in children with CD compared to Ctr or even to UC. Additionally, the relevant increase in C20:0-Cer and C18:1-Cer concentrations were detected in both IBD groups compared to Ctr. The enhanced C24:0-Cer level was observed only in UC, while C18:0-Cer only in the CD group. The highest area under the curve (AUC), specificity, and sensitivity were determined for C16:0-LacCer in CD diagnosis. Our results suggest that the serum LacC16-Cer may be a potential biomarker that distinguishes children with IBD from healthy controls and differentiates IBD subtypes. In addition, C20:0-Cer and C18:0-Cer levels also seem to be closely connected with IBD.

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A novel combination of serum microRNAs for the detection of early gastric cancer

Gastric Cancer ◽

10.1007/s10120-021-01161-0 ◽

2021 ◽

Author(s):

Seiichiro Abe ◽

Juntaro Matsuzaki ◽

Kazuki Sudo ◽

Ichiro Oda ◽

Hitoshi Katai ◽

...

Keyword(s):

Gastric Cancer ◽

Early Gastric Cancer ◽

Expression Profiles ◽

Characteristic Curve ◽

Area Under The Curve ◽

Cancer Center ◽

Serum Samples ◽

Retrospective Case ◽

Serum Mirnas ◽

Validation Set

Abstract Background The aim of this study was to identify serum miRNAs that discriminate early gastric cancer (EGC) samples from non-cancer controls using a large cohort. Methods This retrospective case–control study included 1417 serum samples from patients with EGC (seen at the National Cancer Center Hospital in Tokyo between 2008 and 2012) and 1417 age- and gender-matched non-cancer controls. The samples were randomly assigned to discovery and validation sets and the miRNA expression profiles of whole serum samples were comprehensively evaluated using a highly sensitive DNA chip (3D-Gene®) designed to detect 2565 miRNA sequences. Diagnostic models were constructed using the levels of several miRNAs in the discovery set, and the diagnostic performance of the model was evaluated in the validation set. Results The discovery set consisted of 708 samples from EGC patients and 709 samples from non-cancer controls, and the validation set consisted of 709 samples from EGC patients and 708 samples from non-cancer controls. The diagnostic EGC index was constructed using four miRNAs (miR-4257, miR-6785-5p, miR-187-5p, and miR-5739). In the discovery set, a receiver operating characteristic curve analysis of the EGC index revealed that the area under the curve (AUC) was 0.996 with a sensitivity of 0.983 and a specificity of 0.977. In the validation set, the AUC for the EGC index was 0.998 with a sensitivity of 0.996 and a specificity of 0.953. Conclusions A novel combination of four serum miRNAs could be a useful non-invasive diagnostic biomarker to detect EGC with high accuracy. A multicenter prospective study is ongoing to confirm the present observations.

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Relationship between Plasma Trimethylamine N-Oxide Levels and Renal Dysfunction in Patients with Hypertension

Kidney & Blood Pressure Research ◽

10.1159/000513033 ◽

2021 ◽

pp. 1-12

Author(s):

Jia Zhou ◽

Dingkun Wang ◽

Bingong Li ◽

Xuelian Li ◽

Xingjun Lai ◽

...

Keyword(s):

Blood Pressure ◽

Kidney Disease ◽

Renal Dysfunction ◽

High Performance ◽

Characteristic Curve ◽

Area Under The Curve ◽

Hypertensive Patients ◽

Ckd Stage ◽

Liquid Chromatography Tandem Mass ◽

The Relationship

Introduction: Trimethylamine N-oxide (TMAO) is a metabolite produced by gut bacteria. Although increased TMAO levels have been linked to hypertension (HTN) and chronic kidney disease (CKD) with poor prognosis, no clinical studies have directly addressed the relationship between them. In this study, we investigated the relationship between TMAO and renal dysfunction in hypertensive patients. Methods: We included healthy controls (n = 50), hypertensive patients (n = 46), and hypertensive patients with renal dysfunction (n = 143). Their blood pressure values were taken as the highest measured blood pressure. Renal function was evaluated using the estimated glomerular filtration rate. Plasma TMAO levels were measured using high-performance liquid chromatography tandem mass spectrometry. Results: We found significant differences in plasma TMAO levels among the 3 groups (p < 0.01). The plasma TMAO of patients with HTN was significantly higher than that of healthy people, and the plasma TMAO of patients with HTN complicated by renal dysfunction was significantly higher than either of the other groups. Patients in the highest TMAO quartile were at a higher risk of developing CKD stage 5 than those in the lowest quartile. In the receiver operating characteristic curve, the area under the curve of TMAO combined with β 2-macroglobulin for predicting renal dysfunction in patients with HTN was 0.85 (95% confidence interval 0.80–0.90). Conclusion: An elevated TMAO level reflects higher levels of HTN and more severe renal dysfunction. TMAO, combined with β 2-macroglobulin levels, may assist in diagnosing CKD in hypertensive patients. Plasma TMAO has predictive value for early kidney disease in hypertensive patients.

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Diagnosis of Primary Aldosteronism by Seated Saline Suppression Test—Variability Between Immunoassay and HPLC-MS/MS

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/clinem/dgz150 ◽

2019 ◽

Vol 105 (3) ◽

pp. e477-e483 ◽

Cited By ~ 2

Author(s):

Moe Thuzar ◽

Karen Young ◽

Ashraf H Ahmed ◽

Greg Ward ◽

Martin Wolley ◽

...

Keyword(s):

Primary Aldosteronism ◽

High Performance ◽

Area Under The Curve ◽

Lower Sensitivity ◽

Operating Characteristics ◽

Chemiluminescence Immunoassay ◽

Plasma Aldosterone Concentration ◽

Salt Loading ◽

Performance Area ◽

Test Variability

Abstract Background In primary aldosteronism (PA), excessive, autonomous secretion of aldosterone is not suppressed by salt loading or fludrocortisone. For seated saline suppression testing (SSST), the recommended diagnostic cutoff 4-hour plasma aldosterone concentration (PAC) measured by high-performance liquid chromatography–mass spectrometry (HPLC-MS/MS is 162 pmol/L. Most diagnostic laboratories, however, use immunoassays to measure PAC. The cutoff for SSST using immunoassay is not known. We hypothesized that the cutoff is different between the assays. Methods We analyzed 80 of the 87 SSST tests that were performed during our recent study defining the HPLC-MS/MS cutoff. PA was confirmed in 65 by positive fludrocortisone suppression testing (FST) and/or lateralization on adrenal venous sampling and excluded in 15 by negative FST. PAC was measured by a chemiluminescence immunoassay (PACIA) in the SSST samples using the DiaSorin Liaison XL analyzer, and receiver operating characteristics (ROC) analysis was performed to identify the PACIA cutoff. Results ROC revealed good performance (area under the curve = 0.893; P < .001) of 4-hour postsaline PACIA for diagnosis of PA and an optimal diagnostic cutoff of 171 pmol/L, with sensitivity and specificity of 95.4% and 80.0%, respectively. A higher cutoff of 217 pmol/L improved specificity (86.7%) with lower sensitivity (86.2%). PACIA measurements strongly correlated with PAC measured by HPLC-MS (r = 0.94, P < .001). Conclusions A higher diagnostic cutoff for SSST should be employed when PAC is measured by immunoassay rather than HPLC-MS/MS. The results suggest that (i) PA can be excluded if 4-hour PACIA is less than 171 pmol/L, and (ii) PA is highly likely if the PACIA is greater than 217 pmol/L by chemiluminescence immunoassay. A gray zone exists between the cutoffs of 171 and 217 pmol/L, likely reflecting a lower specificity of immunoassay.

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Exploring serum metabolic markers for the discrimination of ccRCC from renal angiomyolipoma by metabolomics

Biomarkers in Medicine ◽

10.2217/bmm-2019-0215 ◽

2020 ◽

Vol 14 (8) ◽

pp. 675-682

Author(s):

Mingfeng Xiang ◽

Feng Du ◽

Jing Dai ◽

Ling Chen ◽

Ruijin Geng ◽

...

Keyword(s):

Differential Diagnosis ◽

Uric Acid ◽

Characteristic Curve ◽

Area Under The Curve ◽

Serum Levels ◽

Resonance Spectroscopy ◽

Renal Angiomyolipoma ◽

Serum Samples ◽

Metabolic Markers ◽

Cell Renal Carcinoma

Aim: The discrimination of renal cell carcinoma from renal angiomyolipoma (RAML) is crucial for the effective treatment of each. Materials & methods: Serum samples were analyzed by nuclear magnetic resonance spectroscopy-based metabolomics and a number of metabolites were further quantified by HPLC–UV. Results: Clear-cell renal carcinoma (ccRCC) was characterized by drastic disruptions in energy, amino acids, creatinine and uric acid metabolic pathways. A logistic model for the differential diagnosis of RAML from ccRCC was established using the combination of serum levels of uric acid, the ratio of uric acid to hypoxanthine and the ratio of hypoxanthine to creatinine as variables with area under the curve of the receiver operating characteristic curve value of 0.907. Conclusion: Alterations in serum purine metabolites may be used as potential metabolic markers for the differential diagnosis of ccRCC and RAML.

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piRNA-823 Is a Unique Potential Diagnostic Non-Invasive Biomarker in Colorectal Cancer Patients

Genes ◽

10.3390/genes12040598 ◽

2021 ◽

Vol 12 (4) ◽

pp. 598

Author(s):

Norhan A. Sabbah ◽

Wael M. Abdalla ◽

Walid A. Mawla ◽

Nagla AbdAlMonem ◽

Amal F. Gharib ◽

...

Keyword(s):

Colorectal Cancer ◽

Cancer Patients ◽

Characteristic Curve ◽

Area Under The Curve ◽

Serum Levels ◽

P Element ◽

Serum Samples ◽

Expression Levels ◽

Non Invasive ◽

Colorectal Cancer Patients

Early detection of colorectal cancer (CRC) is the most important factor in deciding its prognosis, so the need to develop an accurate screening test is a must. P-element induced wimpy testis (PIWI) RNA-823 (piR-823) is one of the first piRNAs recognized to be linked to malignancy. We aimed to investigate the expression levels of piR-823 in both serum and tissues of colorectal cancer patients and the ability to use its serum level as a non-invasive diagnostic biomarker to detect colorectal cancer. We determined piR-823 expression levels in 84 serum samples of CRC patients, 75 serum samples of healthy controls, and biological specimens obtained from the 84 patients with colorectal cancer from both the tumor tissues and the normal neighboring tissues using quantitative real-time reverse transcriptase-PCR. We showed that piR-823 had significantly higher serum and tissue expression levels in CRC patients compared to the controls. We observed a significant positive correlation between piR-823 serum levels and the staging of CRC, with significantly higher levels exhibiting advanced stages of CRC (III and IV). This translates into poorer differentiation and lymph node metastasis. The receiver operating characteristic curve (ROC curve) test showed 83.3% sensitivity and 89.3% specificity at a cut-off value of >5.98-fold change, with an area under the curve of 0.933 (p < 0.0001) concerning the ability of piR-823 in diagnosing patients with colorectal carcinoma. piR-823 expression is upregulated in colorectal cancer patients’ serum and tissues, and it can be used as a diagnostic noninvasive biomarker for CRC.

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High Concentrations of Measles Neutralizing Antibodies and High-Avidity Measles IgG Accurately Identify Measles Reinfection Cases

Clinical and Vaccine Immunology ◽

10.1128/cvi.00268-16 ◽

2016 ◽

Vol 23 (8) ◽

pp. 707-716 ◽

Cited By ~ 27

Author(s):

Sun B. Sowers ◽

Jennifer S. Rota ◽

Carole J. Hickman ◽

Sara Mercader ◽

Susan Redd ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Natural Infection ◽

Characteristic Curve ◽

Area Under The Curve ◽

Neutralizing Antibody ◽

The United States ◽

High Avidity ◽

Serum Samples ◽

Negative Results ◽

High Concentrations

ABSTRACTIn the United States, approximately 9% of the measles cases reported from 2012 to 2014 occurred in vaccinated individuals. Laboratory confirmation of measles in vaccinated individuals is challenging since IgM assays can give inconclusive results. Although a positive reverse transcription (RT)-PCR assay result from an appropriately timed specimen can provide confirmation, negative results may not rule out a highly suspicious case. Detection of high-avidity measles IgG in serum samples provides laboratory evidence of a past immunologic response to measles from natural infection or immunization. High concentrations of measles neutralizing antibody have been observed by plaque reduction neutralization (PRN) assays among confirmed measles cases with high-avidity IgG, referred to here as reinfection cases (RICs). In this study, we evaluated the utility of measuring levels of measles neutralizing antibody to distinguish RICs from noncases by receiver operating characteristic curve analysis. Single and paired serum samples with high-avidity measles IgG from suspected measles cases submitted to the CDC for routine surveillance were used for the analysis. The RICs were confirmed by a 4-fold rise in PRN titer or by RT-quantitative PCR (RT-qPCR) assay, while the noncases were negative by both assays. Discrimination accuracy was high with serum samples collected ≥3 days after rash onset (area under the curve, 0.953; 95% confidence interval [CI], 0.854 to 0.993). Measles neutralizing antibody concentrations of ≥40,000 mIU/ml identified RICs with 90% sensitivity (95% CI, 74 to 98%) and 100% specificity (95% CI, 82 to 100%). Therefore, when serological or RT-qPCR results are unavailable or inconclusive, suspected measles cases with high-avidity measles IgG can be confirmed as RICs by measles neutralizing antibody concentrations of ≥40,000 mIU/ml.

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Fasting Serum Lipid and Dehydroepiandrosterone Sulfate as Important Metabolites for Detecting Isolated Postchallenge Diabetes: Serum Metabolomics via Ultra-High-Performance LC-MS

Clinical Chemistry ◽

10.1373/clinchem.2012.200527 ◽

2013 ◽

Vol 59 (9) ◽

pp. 1338-1348 ◽

Cited By ~ 34

Author(s):

Liyan Liu ◽

Maoqing Wang ◽

Xue Yang ◽

Mingxin Bi ◽

Lixin Na ◽

...

Keyword(s):

Oleic Acid ◽

Linoleic Acid ◽

Plasma Glucose ◽

High Performance ◽

Large Scale ◽

Area Under The Curve ◽

Dehydroepiandrosterone Sulfate ◽

Fasting Serum ◽

Serum Samples ◽

Postprandial Plasma Glucose

BACKGROUND Isolated postchallenge diabetes (IPD), a subtype of type 2 diabetes mellitus (T2DM) defined as 2-h postprandial plasma glucose ≥200 mg/dL (≥11.1 mmol/L) and fasting plasma glucose (FPG) <108 mg/dL (<6.0 mmol/L), is often overlooked during screening for diabetes on the basis of FPG concentrations. A key challenge is early identification of IPD by the use of fasting serum, which is critical for large-scale diabetes screening. METHODS We applied a nontargeted metabolomic approach using ultra-high-performance liquid chromatography–quadrupole TOF–mass spectrometry (UPLC-QTOF-MS) to analyze serum samples from 51 patients with IPD, 52 with newly diagnosed T2DM, and 49 healthy individuals. We processed metabolite profiles by multivariate analysis to identify potential metabolites, which were further confirmed by tandem MS (MS/MS). We also used GC-MS and ELISA methods to detect potentially important metabolites. A number of independent samples were selected to validate the identified candidates. RESULTS We selected 15 metabolites with a view to distinguishing patients with IPD, whereas 11 were identified with an authentic standard. The selected metabolites included linoleic acid, oleic acid, phospholipids, and dehydroepiandrosterone sulfate (DHEA-S). In IPD samples, significantly higher linoleic and oleic acid (P < 0.001) and lower DHEA-S (P < 0.001) concentrations were observed, compared with controls. The area under the curve from a combination of linoleic acid, oleic acid, and DHEA-S in the validation study was 0.849 for the IPD group. CONCLUSIONS The current study provides useful information to bridge the gaps in our understanding of the metabolic alterations associated with IPD and might facilitate the characterization of patients with IPD by the use of fasting serum.

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Clinical Value and Potential Target of miR-27a-3p in Pulpitis

NeuroImmunoModulation ◽

10.1159/000516136 ◽

2021 ◽

pp. 1-8

Author(s):

Ming Zhou ◽

Chaohong Li

Keyword(s):

Target Gene ◽

Characteristic Curve ◽

Luciferase Reporter ◽

Blood Monocytes ◽

Clinical Value ◽

Serum Samples ◽

Pulp Tissue ◽

Direct Target Gene ◽

High Level ◽

Irreversible Pulpitis

Introduction: This study investigated the clinical values of miR-27a-3p for pulpitis patients, and its association with TLR4. Methods: Sixty-six patients with pulpitis and 34 cases without pulpitis were recruited; the pulp tissue and serum samples were collected from each participant. Real-time polymerase chain reaction was used for measurement of gene expression levels. The diagnosis values were assessed by the receiver operating characteristic curve. The target gene of miR-27a-3p was confirmed by the luciferase reporter assay. Results: MiR-27a-3p was downregulated in both serum and pulp tissue of pulpitis patients. MiR-27a-3p could distinguish pulpitis patients from healthy controls and might be a predictor for the development of irreversible pulpitis. A high level of TLR4 was also detected in both peripheral blood monocytes and pulp tissues from pulpitis patients and showed a negative association with the miR-27a-3p level. TLR4 was a direct target gene of miR-27a-3p. Discussion/Conclusion: MiR-27a-3p might be a promising biomarker for the diagnosis of pulpitis and predict the development of irreversible pulpitis. MiR-27a-3p might be involved in the pathogenesis of pulpitis via targeting TLR4.

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Analysis of serum homocysteine in the laboratory practice - comparison of the direct chemiluminescence immunoassay and high performance liquid chromatography coupled with fluorescent detection

Biochemia Medica ◽

10.11613/bm.2020.030703 ◽

2020 ◽

Vol 30 (3) ◽

pp. 439-445

Author(s):

Łukasz Paprotny ◽

Dorota Wianowska ◽

Magdalena Izdebska ◽

Agnieszka Celejewska ◽

Dorota Szewczak ◽

...

Keyword(s):

High Performance Liquid Chromatography ◽

Regression Analysis ◽

Liquid Chromatography ◽

High Performance ◽

Reference Method ◽

Clinical Analysis ◽

Fluorescent Detection ◽

Reference Ranges ◽

Serum Samples ◽

Chemiluminescence Immunoassay

Introduction: Effective diagnosis of cardiovascular diseases requires the right tools to be used enabling selective and sensitive analysis of their biomarkers. One of them is homocysteine (Hcy), nowadays determined by immunoassays and chromatographic methods. This study aims to compare the results obtained by direct chemiluminescence immunoassay (CLIA) and high performance liquid chromatography with fluorescent detection (HPLC-FD) using commercial kits. Materials and methods: Homocysteine concentration was determined in serum samples obtained from 101 individuals, using Atellica IM HCY (Siemens Healthineers, Erlangen, Germany) and HCY in plasma/serum – HPLC-FD (Chromsystems Instruments & Chemicals GmbH, Gräfelfing, Germany) tests validated for routine analysis. The latter was applied as a reference method. The comparability and agreement between the tested methods were evaluated using the Passing-Bablok (PB) regression analysis and the Bland-Altman (BA) method of the differences analysis. Results: Studies showed that CLIA gives higher Hcy concentrations (15.7 ± 4.14 μmol/L). Passing-Bablok regression analysis of the results obtained with CLIA (y) compared with HPLC-FD (x) yielded an intercept of 0.22 (95%CI: - 2.16 to 2.46) and slope of 1.58 (95%CI: 1.33 to 1.87). Bland-Altman analysis demonstrated a systematic positive bias for CLIA of 5.85 ± 2.77 μmol/L. Conclusions: Methods disagreement precludes their interchangeability. Lower Hcy values by HPLC-FD result from its greater selectivity. High performance liquid chromatography with fluorescent detection should be considered as preferential method for analysing Hcy in blood serum as well as the recommended reference method for routine clinical analysis. This fact, however, imposes the need to establish new reference ranges.

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