scholarly journals Immunoglobulin E Detection Method Based on Cascade Enzymatic Reaction Utilizing Portable Personal Glucose Meter

Sensors ◽  
2021 ◽  
Vol 21 (19) ◽  
pp. 6396
Author(s):  
Hyogu Han ◽  
Junhyun Park ◽  
Jun Ki Ahn
Keyword(s):  
Detection Method ◽  
Immunoglobulin E ◽  
Limit Of Detection ◽  
Enzymatic Reaction ◽  
High Specificity ◽  
Sandwich Assay ◽  
Elisa Kit ◽  
Personal Glucose Meter ◽  
Diagnostic Capability ◽  
Glucose Meter

We herein describe a cascade enzymatic reaction (CER)-based IgE detection method utilizing a personal glucose meter (PGM), which relies on alkaline phosphatase (ALP) activity that regulates the amount of adenosine triphosphate (ATP). The amount of sandwich assay complex is determined according to the presence or absence of the target IgE. Additionally, the ALP in the sandwich assay catalyzes the dephosphorylation of ATP, a substrate of CER, which results in the changes in glucose level. By employing this principle, IgE was reliably detected at a concentration as low as ca. 29.6 ng/mL with high specificity toward various proteins. Importantly, the limit of detection (LOD) of this portable PGM-based approach was comparable to currently commercialized ELISA kit without expensive and bulky analysis equipment as well as complexed washing step. Finally, the diagnostic capability of this method was also successfully verified by reliably detecting IgE present in a real human serum sample with an excellent recovery ratio within 100 ± 6%.

scholarly journals Development of a DNA-Based Detection Method for Cocos Nucifera Using TaqMan™ Real-Time PCR

Foods ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 332
Author(s):  
Jasmin Wrage ◽  
Oxana Kleyner ◽  
Sascha Rohn ◽  
Jürgen Kuballa
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Detection Method ◽  
Immunoglobulin E ◽  
Limit Of Detection ◽  
Cocos Nucifera ◽  
Cross Reactivity ◽  
Pcr Assay ◽  
Probe System ◽  
Ige Mediated

So far, only a few cases of immunoglobulin E (IgE)-mediated coconut allergies have been described in the literature. Due to a growing consumption of coconut-containing foods in occidental countries, the number of coconut allergies may also increase. As there is no causative immunotherapy in clinical routine, appropriate food labelling is particularly important, also with regard to cross-contamination, to prevent serious health consequences. The purpose of this study was to develop a DNA-based detection method for coconut (Cocos nucifera). Initially, three sets of coconut-specific primers were designed and tested. A TaqMan™ probe was then developed to identify and quantify coconut by real-time PCR assay. With 27 other plant and animal species, the specificity of the primer/probe system was tested and cross reactivity was excluded. In a dilution series, a limit of detection of 1 pg/µL was determined. Thus, the developed real-time PCR assay is a suitable method to detect coconut in food.

A portable and antibody-free sandwich assay for determination of chloramphenicol in food based on a personal glucose meter

2015 ◽  
Vol 407 (9) ◽  
pp. 2499-2507 ◽  
Author(s):  
Si Chen ◽  
Ning Gan ◽  
Huairong Zhang ◽  
Futao Hu ◽  
Tianhua Li ◽  
...  
Keyword(s):  
Sandwich Assay ◽  
Personal Glucose Meter ◽  
Glucose Meter

scholarly journals Immobilization-Free Electrochemical Sensor Coupled with a Graphene-Oxide-Based Aptasensor for Glycated Albumin Detection

Biosensors ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 85
Author(s):  
Wassa Waiwinya ◽  
Thitirat Putnin ◽  
Dechnarong Pimalai ◽  
Wireeya Chawjiraphan ◽  
Nuankanya Sathirapongsasuti ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Electrochemical Sensor ◽  
Detection Method ◽  
Limit Of Detection ◽  
Glycated Albumin ◽  
Cost Effective ◽  
Logarithmic Scale ◽  
Electrochemical Aptasensor ◽  
Alternative Approach

An immobilization-free electrochemical sensor coupled with a graphene oxide (GO)-based aptasensor was developed for glycated human serum albumin (GHSA) detection. The concentration of GHSA was monitored by measuring the electrochemical response of free GO and aptamer-bound GO in the presence of glycated albumin; their currents served as the analytical signals. The electrochemical aptasensor exhibited good performance with a base-10 logarithmic scale. The calibration curve was achieved in the range of 0.01–50 µg/mL. The limit of detection (LOD) was 8.70 ng/mL. The developed method was considered a one-drop measurement process because a fabrication step and the probe-immobilization process were not required. This simple sensor offers a cost-effective, rapid, and sensitive detection method, and could be an alternative approach for determination of GHSA levels.

scholarly journals Correction to “DNAzyme Amplified Aptasensing Platform for Ochratoxin A Detection Using a Personal Glucose Meter”

2021 ◽  
Author(s):  
Songbai Zhang ◽  
Yunxia Luan ◽  
Mengyi Xiong ◽  
Jingjing Zhang ◽  
Ryan Lake ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Personal Glucose Meter ◽  
Glucose Meter

Cds nanocrystal enhanced TiO2photoelectrochemical aptasensor for sensitive detection of cytochrome c

2021 ◽  
Vol 11 (11) ◽  
pp. 1774-1780
Author(s):  
Shanji Fan ◽  
Hong Huang ◽  
Hong Chen ◽  
Jiachi Xu ◽  
Zecheng Hu ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Limit Of Detection ◽  
Specific Binding ◽  
High Sensitivity ◽  
High Specificity ◽  
Linear Range ◽  
Current Intensity ◽  
Sensitive Detection ◽  
Layer Adsorption ◽  
Ionic Layer

A CdS nanocrystal enhanced TiO2 nanotubes (CdS@TiO2 NATs) photoelectrode was prepared via successive ionic layer adsorption and reaction (SILAR) of CdS on the surface of TiO2 NATs. A HS-aptamer owing a specific binding toward cytochrome c was modified onto the CdS@TiO2 NATs, which resulting a decrease in the photoelectrical current intensity. Cytochrome c is therefore quantified based on the decrease in photoelectrical current. High specificity and high sensitivity were obtained with a linear range from 3 pM to 80 nM, and a limit of detection of 2.53 pM.

A Simple and Rapid Light-Initiated Chemiluminescence Assay for Quantitation of Artemisia-Specific Immunoglobulin E

2021 ◽  
pp. 1-8
Author(s):  
Dandan Liu ◽  
Bei Zhang ◽  
Lina Zhu ◽  
Lisheng Zheng ◽  
Shaoshen Li ◽  
...  
Keyword(s):  
Food Allergen ◽  
Clinical Guideline ◽  
Immunoglobulin E ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Chemiluminescence Assay ◽  
Limit Of Quantitation ◽  
Specific Immunoglobulin E ◽  
Specific Immunoglobulin

<b><i>Background:</i></b> Light-initiated chemiluminescence assay (LICA) is a homogeneous assay that has been successfully used for the quantitation of food allergen-specific immunoglobulin E (sIgE), but not inhaled allergen-sIgE. Simultaneously, current assays used to detect allergen-sIgE are serum consuming and/or time consuming. Hence, we established a method for the quantitation of <i>Artemisia</i>-sIgE based on LICA and verified its performance according to the clinical guideline documents, laying a foundation for the quantitation of inhaled and food allergen-sIgE in parallel on LICA. <b><i>Methods:</i></b> The assay was established after optimizing the first incubation time and the dilutions of <i>Artemisia</i>-coated chemibeads, biotinylated goat anti-human IgE, and serum. In order to quantitate <i>Artemisia</i>-sIgE, the calibration curve was established with a high positive serum of known concentration. The assay performance was confirmed per the clinical guideline documents. In addition, the correlation between the results of LICA and capture enzyme-linked immunosorbent assay was evaluated. <b><i>Results:</i></b> The developed LICA’s coefficients of variation of repeatability and intermediate precision were 3.20%, 2.14%, and 3.85% and 4.30%, 4.00%, and 4.40%, respectively. The limit of detection was 0.10 kU<sub>A</sub>/L, and the limit of quantitation was 0.11 kU<sub>A</sub>/L. The range of linearity was from 0.27 kU<sub>A</sub>/L to 97.53 kU<sub>A</sub>/L (<i>r</i> = 0.9968). The correlation coefficient (<i>r</i>) for the correlation analysis between results of LICA and capture ELISA was 0.9087. This assay was successfully applied in 64 human serum samples, showing good sensitivity (82.20%) and specificity (100%). <b><i>Conclusion:</i></b> An <i>Artemisia</i>-sIgE quantitation assay based on LICA was successfully established. Its performance satisfied the clinical requirements and could be widely used in clinical laboratories.

Bioinspired DNA–Inorganic Hybrid Nanoflowers Combined with a Personal Glucose Meter for Onsite Detection of miRNA

2018 ◽  
Vol 10 (49) ◽  
pp. 42050-42057 ◽  
Author(s):  
Tingting Wu ◽  
Yuemeng Yang ◽  
Yu Cao ◽  
Yongchao Song ◽  
Li-Ping Xu ◽  
...  
Keyword(s):  
Inorganic Hybrid ◽  
Personal Glucose Meter ◽  
Glucose Meter

scholarly journals SARS-CoV-2 Direct Detection Without RNA Isolation With Loop-Mediated Isothermal Amplification (LAMP) and CRISPR-Cas12

2021 ◽  
Vol 8 ◽  
Author(s):  
Alfredo Garcia-Venzor ◽  
Bertha Rueda-Zarazua ◽  
Eduardo Marquez-Garcia ◽  
Vilma Maldonado ◽  
Angelica Moncada-Morales ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Direct Detection ◽  
Direct Method ◽  
Rna Extraction ◽  
Rna Isolation ◽  
High Sensitivity ◽  
High Specificity ◽  
Isothermal Amplification ◽  
Clinical Samples ◽  
Loop Mediated Isothermal Amplification

As to date, more than 49 million confirmed cases of Coronavirus Disease 19 (COVID-19) have been reported worldwide. Current diagnostic protocols use qRT-PCR for viral RNA detection, which is expensive and requires sophisticated equipment, trained personnel and previous RNA extraction. For this reason, we need a faster, direct and more versatile detection method for better epidemiological management of the COVID-19 outbreak. In this work, we propose a direct method without RNA extraction, based on the Loop-mediated isothermal amplification (LAMP) and Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR associated protein (CRISPR-Cas12) technique that allows the fast detection of SARS-CoV-2 from patient samples with high sensitivity and specificity. We obtained a limit of detection of 16 copies/μL with high specificity and at an affordable cost. The diagnostic test readout can be done with a real-time PCR thermocycler or with the naked eye in a blue-light transilluminator. Our method has been evaluated on a small set of clinical samples with promising results.

scholarly journals A Highly Sensitive Cell-Based TLR Reporter Platform for the Specific Detection of Bacterial TLR Ligands

2022 ◽  
Vol 12 ◽  
Author(s):  
Katharina Radakovics ◽  
Claire Battin ◽  
Judith Leitner ◽  
Sabine Geiselhart ◽  
Wolfgang Paster ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Ectopic Expression ◽  
High Sensitivity ◽  
High Specificity ◽  
Reporter System ◽  
Specificity And Sensitivity ◽  
Highly Sensitive ◽  
Definition Of ◽  
The Individual ◽  
Allergen Extracts

Toll-like receptors (TLRs) are primary pattern recognition receptors (PRRs), which recognize conserved microbial components. They play important roles in innate immunity but also in the initiation of adaptive immune responses. Impurities containing TLR ligands are a frequent problem in research but also for the production of therapeutics since TLR ligands can exert strong immunomodulatory properties even in minute amounts. Consequently, there is a need for sensitive tools to detect TLR ligands with high sensitivity and specificity. Here we describe the development of a platform based on a highly sensitive NF-κB::eGFP reporter Jurkat JE6-1 T cell line for the detection of TLR ligands. Ectopic expression of TLRs and their coreceptors and CRISPR/Cas9-mediated deletion of endogenously expressed TLRs was deployed to generate reporter cell lines selectively expressing functional human TLR2/1, TLR2/6, TLR4 or TLR5 complexes. Using well-defined agonists for the respective TLR complexes we could demonstrate high specificity and sensitivity of the individual reporter lines. The limit of detection for LPS was below 1 pg/mL and ligands for TLR2/1 (Pam3CSK4), TLR2/6 (Fsl-1) and TLR5 (flagellin) were detected at concentrations as low as 1.0 ng/mL, 0.2 ng/mL and 10 pg/mL, respectively. We showed that the JE6-1 TLR reporter cells have the utility to characterize different commercially available TLR ligands as well as more complex samples like bacterially expressed proteins or allergen extracts. Impurities in preparations of microbial compounds as well as the lack of specificity of detection systems can lead to erroneous results and currently there is no consensus regarding the involvement of TLRs in the recognition of several molecules with proposed immunostimulatory functions. This reporter system represents a highly suitable tool for the definition of structural requirements for agonists of distinct TLR complexes.

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