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

2010 ◽  
Vol 17 (3) ◽  
pp. 335-341 ◽  
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.

Serum metabolomics reveals dysregulation and diagnostic potential of oxylipins in tumor-induced osteomalacia

2021 ◽  
Author(s):  
Yiyi Gong ◽  
Xiaolin Ni ◽  
Chenxi Jin ◽  
Xiang Li ◽  
Yujie Wang ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Fibroblast Growth Factor 23 ◽  
Tumor Resection ◽  
High Sensitivity ◽  
Leukotriene B4 ◽  
Arachidonic Acid Metabolism ◽  
Serum Samples ◽  
Diagnostic Potential ◽  
Liquid Chromatography Tandem Mass ◽  
Diagnostic Panel

Abstract Context Excessive production of fibroblast growth factor 23 (FGF23) by tumor was considered as the main pathogenesis in tumor-induced osteomalacia (TIO). Despite its importance to comprehensive understanding of pathogenesis and diagnosis, the regulation of systemic metabolism in TIO remains unclear. Objectives We aimed to systematically characterize the metabolome alteration associated with TIO. Methods By means of liquid chromatography-tandem mass spectrometry (LC-MS) based metabolomics, we analyzed the metabolic profile from 96 serum samples (32 initial diagnosis TIO patients, pairwise samples after tumor resection and 32 matched healthy control subjects). In order to screen and evaluate potential biomarkers, statistical analyses, pathway enrichment and receiver operating characteristic (ROC) were performed. Results Metabolomic profiling revealed distinct alterations between TIO and HC cohort. Differential metabolites were screened and conducted to functional clustering and annotation. Significantly enriched pathway was found involved in arachidonic acid metabolism. A combination of 5 oxylipins, 4-HDoHE, leukotriene B4, 5-HETE, 17-HETE and 9,10,13-TriHOME, demonstrated a high sensitivity and specificity panel for TIO prediction screened by random forest (RF) algorithm (AUC=0.951, 95% confidence interval, CI 0.827-1). Supported vector machine (SVM) model and partial least-squares (PLS) model were conducted to validate the predictive capabilities of the diagnostic panel. Conclusions Metabolite profiling of TIO altered significant compared with HC. A high sensitivity and specificity panel with 5 oxylipins were tested as diagnostic predictor. For the first time, we provide the global profile of metabolomes and identify potential diagnostic biomarkers of TIO. The present work may offer novel insights into the pathogenesis of TIO.

scholarly journals Sensitive detection of multiple islet autoantibodies in type 1 diabetes using small sample volumes by agglutination-PCR

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0242049
Author(s):  
Felipe de Jesus Cortez ◽  
David Gebhart ◽  
Peter V. Robinson ◽  
David Seftel ◽  
Narges Pourmandi ◽  
...  
Keyword(s):  
Type 1 Diabetes ◽  
Sensitivity And Specificity ◽  
High Sensitivity ◽  
Small Sample ◽  
Insulin Autoantibodies ◽  
Islet Autoantibodies ◽  
Antibody Detection ◽  
Clinical Sensitivity ◽  
Analysis Platform

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.

scholarly journals Antibody detection by agglutination–PCR (ADAP) enables early diagnosis of HIV infection by oral fluid analysis

2018 ◽  
Vol 115 (6) ◽  
pp. 1250-1255 ◽  
Author(s):  
Cheng-ting Tsai ◽  
Peter V. Robinson ◽  
Felipe de Jesus Cortez ◽  
Maria L. B. Elma ◽  
David Seftel ◽  
...  
Keyword(s):  
Oral Fluid ◽  
Detection Threshold ◽  
Population Based ◽  
Antibody Detection ◽  
Analytical Sensitivity ◽  
Sample Collection ◽  
Clinical Sensitivity ◽  
Fluid Analysis ◽  
Hiv Antibodies ◽  
Anti Hiv

Oral fluid (OF) is a highly effective substrate for population-based HIV screening efforts, as it is noninfectious and significantly easier to collect than blood. However, anti-HIV antibodies are found at far lower concentrations in OF compared with blood, leading to poor sensitivity and a longer period of time from infection to detection threshold. Thus, despite its inherent advantages in sample collection, OF is not widely used for population screening. Here we report the development of an HIV OF assay based on Antibody Detection by Agglutination–PCR (ADAP) technology. This assay is 1,000–10,000 times more analytically sensitive than clinical enzyme-linked immunoassays (EIAs), displaying both 100% clinical sensitivity and 100% specificity for detecting HIV antibodies within OF samples. We show that the enhanced analytical sensitivity enables this assay to correctly identify HIV-infected individuals otherwise missed by current OF assays. We envision that the attributes of this improved HIV OF assay can increase testing rates of at-risk individuals while enabling diagnosis and treatment at an earlier time point.

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

2011 ◽  
Vol 44 (13) ◽  
pp. S313
Author(s):  
Saeed Khalili ◽  
Rasaee Mohamad Javad ◽  
Mousavi Seyed Latif ◽  
Jafar Amani ◽  
Abolfazl Jahangiri
Keyword(s):  
Sensitivity And Specificity ◽  
In Silico ◽  
High Sensitivity ◽  
Antibody Detection

scholarly journals Toxoplasma gondii Recombinant antigen AMA1: Diagnostic Utility of Protein Fragments for the Detection of IgG and IgM Antibodies

Pathogens ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 43 ◽  
Author(s):  
Bartłomiej Ferra ◽  
Lucyna Holec-Gąsior ◽  
Justyna Gatkowska ◽  
Bożena Dziadek ◽  
Katarzyna Dzitko
Keyword(s):  
Toxoplasma Gondii ◽  
Expression System ◽  
High Sensitivity ◽  
Recombinant Antigen ◽  
Full Length ◽  
Diagnostic Utility ◽  
Intermediate Hosts ◽  
Serological Tests ◽  
Apical Membrane Antigen 1 ◽  
Diagnostic Potential

Toxoplasma gondii is an important zoonotic protozoan that infects a wide variety of vertebrates as intermediate hosts. For this reason, the diagnosis of this disease is very important and requires continuous improvement. One possibility is to use recombinant antigens in serological tests. Apical membrane antigen 1 (AMA1), a protein located in specific secretory organelles (micronemes) of T. gondii, is very interesting in regard to its potential diagnostic utility. In the present study, we attempted to identify a fragment of the AMA1 protein with a high sensitivity and specificity for the serological diagnosis of human toxoplasmosis. The full-length AMA1 and two different fragments (AMA1N and AMA1C) were produced using an Escherichia coli expression system. After purification by metal affinity chromatography, recombinant proteins were tested for their utility as antigens in enzyme-linked immunosorbent assays (ELISAs) for the detection of IgG and IgM anti-T. gondii antibodies in human and mouse immune sera. Our data demonstrate that the full-length AMA1 recombinant antigen (corresponding to amino acid residues 67–569 of the native protein) has a better diagnostic potential than its N- or C-terminal fragments. This recombinant protein strongly interacts with specific anti-T. gondii IgG (99.4%) and IgM (80.0%) antibodies, and may be used for developing new tools for diagnostics of toxoplasmosis.

Time-resolved immunofluorescence: a sensitive and specific assay for anti-HIV antibody detection in human sera

1987 ◽  
Vol 16 (4) ◽  
pp. 303-315 ◽  
Author(s):  
A. Aceti ◽  
F. Titti ◽  
P. Verani ◽  
S. Buttò ◽  
A. Pennica ◽  
...  
Keyword(s):  
Antibody Detection ◽  
Time Resolved ◽  
Specific Assay ◽  
Human Sera ◽  
Anti Hiv

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

Parasitology ◽  
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.

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

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

TO SEPARATELY ASSESS THE DIAGNOSTIC POTENTIAL OF TVS & MRI IN THE TERMS OF SENSITIVITY AND SPECIFICITY BY CORRELATING THE IMAGING FINDINGS WITH HISTOPATHOLOGY

2020 ◽  
Vol 4 (6) ◽  
Author(s):  
Suraj Mathur
Keyword(s):  
Transvaginal Ultrasound ◽  
High Sensitivity ◽  
Imaging Evaluation ◽  
Medical College ◽  
Conventional Mri ◽  
Diagnostic Potential ◽  
Adc Mapping ◽  
Malignant Lesions ◽  
Sensitivity Specificity

This prospective study was done in the Department of Radio diagnosis Govt. Medical College, Kozhikode. A total of 65 patients who were referred to our department with clinical suspicion of endometrial lesions and incidentally detected endometrial lesions on ultrasonography underwent transvaginal ultrasound and subsequent Imaging evaluation of pelvis MRI has very high sensitivity (95%) and specificity (98%) and is almost as accurate (97%) as histopathology in differentiating benign from malignant lesions. Addition of DWI with ADC mapping to conventional MRI increases its accuracy even more. However there is inherent limitation to MRI in detecting carcinoma in situ and micrometastasis. Keywords: TVS, MRI, Sensitivity, Specificity, Histopathology.

Trends in Diagnosis for Active Tuberculosis Using Nanomaterials

2019 ◽  
Vol 26 (11) ◽  
pp. 1946-1959 ◽  
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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