Designing of a multiepitopic antigen for anti-HBV antibody detection with high sensitivity and specificity: an in silico approach

Islet autoantibodies are predominantly measured by radioassay to facilitate risk assessment and diagnosis of type 1 diabetes. However, the reliance on radioactive components, large sample volumes and limited throughput renders radioassay testing costly and challenging. We developed a multiplex analysis platform based on antibody detection by agglutination-PCR (ADAP) for the sample-sparing measurement of GAD, IA-2 and insulin autoantibodies/antibodies in 1 μL serum. The assay was developed and validated in 7 distinct cohorts (n = 858) with the majority of the cohorts blinded prior to analysis. Measurements from the ADAP assay were compared to radioassay to determine correlation, concordance, agreement, clinical sensitivity and specificity. The average overall agreement between ADAP and radioassay was above 91%. The average clinical sensitivity and specificity were 96% and 97%. In the IASP 2018 workshop, ADAP achieved the highest sensitivity of all assays tested at 95% specificity (AS95) rating for GAD and IA-2 autoantibodies and top-tier performance for insulin autoantibodies. Furthermore, ADAP correctly identified 95% high-risk individuals with two or more autoantibodies by radioassay amongst 39 relatives of T1D patients tested. In conclusion, the new ADAP assay can reliably detect the three cardinal islet autoantibodies/antibodies in 1μL serum with high sensitivity. This novel assay may improve pediatric testing compliance and facilitate easier community-wide screening for islet autoantibodies.

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Development of a TaqMan qPCR protocol for detecting Acipenser ruthenus in the Volga headwaters from eDNA samples

Conservation Genetics Resources ◽

10.1007/s12686-020-01128-w ◽

2020 ◽

Vol 12 (3) ◽

pp. 395-397 ◽

Cited By ~ 1

Author(s):

Tamara Schenekar ◽

Martin Schletterer ◽

Steven J. Weiss

Keyword(s):

Sensitivity And Specificity ◽

In Silico ◽

High Sensitivity ◽

In Vitro Tests ◽

Fish Diversity ◽

Upper Volga ◽

Acipenser Ruthenus ◽

Taqman Qpcr

AbstractA recent eDNA-metabarcoding study assessing fish diversity in the Upper Volga catchment did not detect sterlet at any of the sampled stretches, despite recent sightings that suggest its presence. We designed a TaqMan qPCR protocol to test for sterlet in selected eDNA samples from that study. In-silico and in-vitro tests confirm the protocol’s high sensitivity and specificity to sturgeon taxa and potentially paddlefishes. Using this assay, sterlet were not detected in 26 eDNA samples from the Volga headwaters, agreeing with the metabarcoding results.

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Inexpensive Designer Antigen for Anti-HIV Antibody Detection with High Sensitivity and Specificity

Clinical and Vaccine Immunology ◽

10.1128/cvi.00283-09 ◽

2010 ◽

Vol 17 (3) ◽

pp. 335-341 ◽

Cited By ~ 4

Author(s):

Sheikh M. Talha ◽

Teppo Salminen ◽

Deepti A. Chugh ◽

Sathyamangalam Swaminathan ◽

Tero Soukka ◽

...

Keyword(s):

Sensitivity And Specificity ◽

Expression System ◽

High Sensitivity ◽

Antibody Detection ◽

Human Antibody ◽

Time Resolved ◽

Reading Frame ◽

Diagnostic Potential ◽

Hiv Antibodies ◽

Anti Hiv

ABSTRACT A novel recombinant multiepitope protein (MEP) has been designed that consists of four linear, immunodominant, and phylogenetically conserved epitopes, taken from human immunodeficiency virus (HIV)-encoded antigens that are used in many third-generation immunoassay kits. This HIV-MEP has been evaluated for its diagnostic potential in the detection of anti-HIV antibodies in human sera. A synthetic MEP gene encoding these epitopes, joined by flexible peptide linkers in a single open reading frame, was designed and overexpressed in Escherichia coli. The recombinant HIV-MEP was purified using a single affinity step, yielding >20 mg pure protein/liter culture, and used as the coating antigen in an in-house immunoassay. Bound anti-HIV antibodies were detected by highly sensitive time-resolved fluorometry, using europium(III) chelate-labeled anti-human antibody. The sensitivity and specificity of the HIV-MEP were evaluated using Boston Biomedica worldwide HIV performance, HIV seroconversion, and viral coinfection panels and were found to be comparable with those of commercially available anti-HIV enzyme immunoassay (EIA) kits. The careful choice of epitopes, high epitope density, and an E. coli-based expression system, coupled with a simple purification protocol and the use of europium(III) chelate-labeled tracer, provide the capability for the development of an inexpensive diagnostic test with high degrees of sensitivity and specificity.

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A new ELISA and western blot technique based on recombinant TES antigen and/or larval antigen for the detection of toxocariasis in humans

Parasitology ◽

10.1017/s0031182020002085 ◽

2020 ◽

pp. 1-8

Author(s):

Marie-Kristin Raulf ◽

Daniela Jordan ◽

Herbert Auer ◽

Jens M. Warnecke ◽

Bernd Lepenies ◽

...

Keyword(s):

Sensitivity And Specificity ◽

High Reliability ◽

High Sensitivity ◽

Cross Reactivity ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Serological Tests ◽

Western Blots ◽

Blot Technique ◽

Human Sera

Abstract Serological antibody detection by enzyme-linked immunosorbent assay (ELISA)- and immunoblot-based methods constitutes the best indicator of human Toxocara infection. Nevertheless, the availability of serological tests, particularly western blots (WB), evaluated for sensitivity and specificity is limited. Therefore, an Anti-Toxocara-ELISA immunoglobulin g (IgG) prototype (Proto-ELISA) and an Anti-Toxocara-Westernblot (IgG) prototype (Proto-WB) were evaluated by testing 541 human sera pre-determined for Toxocara infection by an established in-house Anti-Toxocara-ELISA (IH-ELISA). To evaluate sensitivity and specificity of the newly developed ELISA and WB prototypes, results were compared to IH-ELISA and a commercial WB (Com-WB). Compared to the IH-ELISA, a sensitivity of 93.1% (229/246) and a specificity of 94.6% (279/295) of the Proto-ELISA with a Cohen's κ of 0.88 were obtained. The sensitivity of the Proto-WB was 76.7% (240/313) and specificity was 99.6% (227/228) with a Cohen's κ of 0.73 compared to those of Com-WB. A comparison to the IH-ELISA revealed 91.5% (225/246) sensitivity and 94.6% (279/295) specificity of the Proto-WB with a Cohen's κ of 0.86. Cross-reactivity was observed for some samples positive for Ascaris and Trichinella spp. in the Proto-ELISA, Proto-WB and Com-WB. Overall, the evaluated ELISA and WB prototypes showed high sensitivity and specificity, indicating high reliability of these newly developed tests.

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Detector-C: A Blood-based IVD with High Sensitivity and Specificity for Early Detection and Screening of Colorectal Cancer

Zeitschrift für Gastroenterologie ◽

10.1055/s-0030-1263628 ◽

2010 ◽

Vol 48 (08) ◽

Author(s):

A Rosenthal ◽

H Köppen ◽

R Musikowski ◽

R Schwanitz ◽

J Behrendt ◽

...

Keyword(s):

Colorectal Cancer ◽

Early Detection ◽

Sensitivity And Specificity ◽

High Sensitivity

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Trends in Diagnosis for Active Tuberculosis Using Nanomaterials

Current Medicinal Chemistry ◽

10.2174/0929867325666180912105617 ◽

2019 ◽

Vol 26 (11) ◽

pp. 1946-1959 ◽

Cited By ~ 3

Author(s):

Le Minh Tu Phan ◽

Lemma Teshome Tufa ◽

Hwa-Jung Kim ◽

Jaebeom Lee ◽

Tae Jung Park

Keyword(s):

Sensitivity And Specificity ◽

High Efficiency ◽

Point Of Care ◽

High Sensitivity ◽

Signs And Symptoms ◽

Diagnostic Methods ◽

Advantages And Disadvantages ◽

Treatment And Prevention ◽

Clinical Tools ◽

Diagnostic Applications

Background:Tuberculosis (TB), one of the leading causes of death worldwide, is difficult to diagnose based only on signs and symptoms. Methods for TB detection are continuously being researched to design novel effective clinical tools for the diagnosis of TB.Objective:This article reviews the methods to diagnose TB at the latent and active stages and to recognize prospective TB diagnostic methods based on nanomaterials.Methods:The current methods for TB diagnosis were reviewed by evaluating their advantages and disadvantages. Furthermore, the trends in TB detection using nanomaterials were discussed regarding their performance capacity for clinical diagnostic applications.Results:Current methods such as microscopy, culture, and tuberculin skin test are still being employed to diagnose TB, however, a highly sensitive point of care tool without false results is still needed. The utilization of nanomaterials to detect the specific TB biomarkers with high sensitivity and specificity can provide a possible strategy to rapidly diagnose TB. Although it is challenging for nanodiagnostic platforms to be assessed in clinical trials, active TB diagnosis using nanomaterials is highly expected to achieve clinical significance for regular application. In addition, aspects and future directions in developing the high-efficiency tools to diagnose active TB using advanced nanomaterials are expounded.Conclusion:This review suggests that nanomaterials have high potential as rapid, costeffective tools to enhance the diagnostic sensitivity and specificity for the accurate diagnosis, treatment, and prevention of TB. Hence, portable nanobiosensors can be alternative effective tests to be exploited globally after clinical trial execution.

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Value of Coronary Calcium Scoring in Symptomatic and Asymptomatic Coronary Artery Disease Patients

Current Medical Imaging Formerly Current Medical Imaging Reviews ◽

10.2174/1573405616666201023142030 ◽

2020 ◽

Vol 16 ◽

Author(s):

Hala T. Salem ◽

Eman A.S. Sabek

Keyword(s):

Coronary Artery Disease ◽

Coronary Artery ◽

Sensitivity And Specificity ◽

High Sensitivity ◽

Coronary Calcium ◽

Risk Estimate ◽

Calcium Scoring ◽

Coronary Calcium Scoring ◽

Framingham Risk ◽

Artery Disease

Aim and Objective: To estimate the relationship between Coronary Calcium Scoring (CCS)and presence of different degrees of obstructive coronary artery disease (CAD) to avoid unnecessary examinations and hence unnecessary radiation exposure and contrast injection. Background: Coronary Calcium Scoring (CCS) is a test uses x-ray equipment to produce pictures of the coronary arteries to determine the degree of its narrowing by the build-up of calcified plaques. Despite the lack of definitive data linking ionizing radiation with cancer, the American Heart Association supports widely that practitioners of Computed tomography Coronary Angiography (CTCA) should keep “patient radiation doses as low as reasonably achievable but consistent with obtaining the desired medical information”. Methods: Data obtained from 275 CTCA examinations were reviewed. Radiation effective doses were estimated for both CCS and CTCA, measures to keep it as low as possible were presented, CCS and Framingham risk estimate were compared to the final results of CTCA to detect sensitivity and specificity of each one in detecting obstructive lesions. Results: CCS is a strong discriminator for obstructive CAD and can with high sensitivity and specificity and correlates well with the degree of obstruction even more than Framingham risk estimate which has high sensitivity and low specificity. Conclusion: CCS helps reducing the effective radiation dose if properly evaluated to skip unnecessary CTCA if obstructive lesions was unlikely, and as a test does not use contrast material, harmful effect on the kidney will be avoided as most of coronary atherosclerotic patients have renal problems.

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Blood-Based Detection of Colorectal Cancer Using Cancer-Specific DNA Methylation Markers

Diagnostics ◽

10.3390/diagnostics11010051 ◽

2020 ◽

Vol 11 (1) ◽

pp. 51

Author(s):

Nam-Yun Cho ◽

Ji-Won Park ◽

Xianyu Wen ◽

Yun-Joo Shin ◽

Jun-Kyu Kang ◽

...

Keyword(s):

Colorectal Cancer ◽

Dna Methylation ◽

Sensitivity And Specificity ◽

Stage Iv ◽

High Sensitivity ◽

Liquid Biopsies ◽

Blood Leukocytes ◽

Marker Panel ◽

A Genome ◽

Methylight Assay

Cancer tissues have characteristic DNA methylation profiles compared with their corresponding normal tissues that can be utilized for cancer diagnosis with liquid biopsy. Using a genome-scale DNA methylation approach, we sought to identify a panel of DNA methylation markers specific for cell-free DNA (cfDNA) from patients with colorectal cancer (CRC). By comparing DNA methylomes between CRC and normal mucosal tissues or blood leukocytes, we identified eight cancer-specific methylated loci (ADGRB1, ANKRD13, FAM123A, GLI3, PCDHG, PPP1R16B, SLIT3, and TMEM90B) and developed a five-marker panel (FAM123A, GLI3, PPP1R16B, SLIT3, and TMEM90B) that detected CRC in liquid biopsies with a high sensitivity and specificity with a droplet digital MethyLight assay. In a set of cfDNA samples from CRC patients (n = 117) and healthy volunteers (n = 60), a panel of five markers on the platform of the droplet digital MethyLight assay detected stages I–III and stage IV CRCs with sensitivities of 45.9% and 95.7%, respectively, and a specificity of 95.0%. The number of detected markers was correlated with the cancer stage, perineural invasion, lymphatic emboli, and venous invasion. Our five-marker panel with the droplet digital MethyLight assay showed a high sensitivity and specificity for the detection of CRC with cfDNA samples from patients with metastatic CRC.

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