Feasibility of Immunodiagnostic Devices for the Detection of Ricin, Amanitin, and T-2 Toxin in Food

2005 ◽  
Vol 68 (6) ◽  
pp. 1294-1301 ◽  
Author(s):  
ERIC A. E. GARBER ◽  
ROBERT M. EPPLEY ◽  
MICHAEL E. STACK ◽  
MICHAEL A. McLAUGHLIN ◽  
DOUGLAS L. PARK

Qualitative and quantitative comparisons were conducted of commercially available immunodiagnostic devices for the detection of three select agents with oral LD50 values ≥0.1 mg/kg of body weight. Ricin (oral LD50 > 1 mg/kg), amanitin (oral LD50 approximately 0.1 mg/kg), and T-2 toxin (oral LD50 > 1 mg/kg) were spiked into beverages, produce, dairy, and baked goods and assayed using commercially available enzyme-linked immunosorbent assays (ELISAs) and lateral flow devices. In all cases, the commercial diagnostic kits successfully detected all three select agents at concentrations below what might be a health concern. The considerable difference between the limit of detection of the immunodiagnostic devices employed (typically ≤0.020 μg/g) and the amount of the select agent necessary to pose a health threat in a single serving of food facilitated the design of protocols for the high throughput screening of food samples. These protocols entailed simple extraction methods followed by sample dilution. Lateral flow devices and sandwich ELISAs for the detection of ricin had no significant background problems due to the food matrices. Competitive ELISAs, which typically have unacceptably high background reactions with food samples, successfully detected amanitin and T-2 toxin.

2018 ◽  
Vol 113 ◽  
pp. 95-100 ◽  
Author(s):  
Ioannis N. Katis ◽  
Peijun J.W. He ◽  
Robert W. Eason ◽  
Collin L. Sones

Foods ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 281 ◽  
Author(s):  
Mingfei Pan ◽  
Tianyu Ma ◽  
Jingying Yang ◽  
Shijie Li ◽  
Shengmiao Liu ◽  
...  

This paper describes the development of lateral flow immunochromatographic assays (ICAs) using colloidal Au sphere (SP) and nanorods (NRs) as signal markers for the determination of zearalenone (ZEN) in cereals. The developed ICAs can detect the analyte ZEN within a short time (10 min), and achieve lower limit of detection (LOD). This is the first time that the AuNRs are used as signal probe in immune test strip for ZEN detection. For colloidal AuSP immunochromatographic analysis (AuSP-ICA), the LODs in solution and spiked cereal sample were 5.0 μg L−1 and 60 μg kg−1, and for AuNRs immunochromatographic analysis (AuNRs-ICA) the two LODs achieved 3.0 μg L−1 and 40 μg kg−1, respectively. These two proposed ICAs have minor cross-reaction to the structural analogs of ZEN, and no cross-reactivity with aflatoxin B1, T-2 toxin, ochratoxin A, deoxynivalenol, fumonisin B1. Both of the developed ICAs can specifically and sensitively detect ZEN in cereals, providing an effective strategy for rapid screening and detection of ZEN in a large number of food samples.


2021 ◽  
Author(s):  
Nol Salcedo ◽  
Brena F Sena ◽  
Xiying Qu ◽  
Bobby Brooke Herrera

Human transmission of SARS-CoV-2 and emergent variants of concern has continued to occur globally, despite mass vaccination campaigns. Public health strategies to reduce virus spread should therefore rely, in part, on frequent screening with rapid, inexpensive, and sensitive tests. We evaluated two digitally integrated rapid tests and assessed their performance using stored nasal swab specimens collected from individuals with or without COVID-19. An isothermal amplification assay combined with a lateral flow test had a limit of detection of 10 RNA copies per reaction, and a positive percent agreement (PPA)/negative percent agreement (NPA) during the asymptomatic and symptomatic phases of 100%/100% and 95.83/100%, respectively. Comparatively, an antigen-based lateral flow test, had a limit of detection of 30,000 copies, and a PPA/NPA during the asymptomatic and symptomatic phases of 82.86%/98.68% and 91.67/100%, respectively. Both the isothermal amplification and antigen-based lateral flow tests had optimized detection of SARS-CoV-2 during the peak period of transmission; however, the antigen-based test had reduced sensitivity in clinical samples with qPCR Ct values greater than 29.8. Low-cost, high-throughput screening enabled by isothermal amplification or antigen-based techniques have value for outbreak control.


Author(s):  
Carla Eiras

Interleukin-6 (IL-6) is a multifunctional cytokine and high bloodstream levels of which have been associated with severe inflammatory diseases, such as dengue fever, sepsis, various cancers, and visceral leishmaniasis (VL). Rapid tests for the quantification of IL-6 would be of great assistance for the bedside diagnosis and treatment of diseases such as VL. We have developed a lateral flow assay (LFA) for rapid and colorimetric IL-6 detection, consisting of anti-IL-6 antibodies conjugated to gold nanoparticles (AuNPs). The optimal concentration of anti-IL-6 used in the conjugate was determined to be 800.0 μg/mL, based on an aggregation assay using LFA. A linear relationship between IL-6 standard concentration and color intensity was observed after 20 min, with a linear range between 1.25 ng/mL and 9,000 ng/mL. The limit of detection for this method was estimated a t0.38 ng/mL. The concentration of IL-6 in five patients with severe VL was measured using LFA, and the results were consistent with those obtained using the cytometric bead array (CBA) method. A thorough analysis of the LFA membranes’ surface morphology, before and after sample contact, was performed using atomic force microscopy (AFM).The prototype described here is still being tested and improved, but this LFA will undoubtedly be of great help in the clinical quantification of IL-6.


Author(s):  
Carla Eiras

Interleukin-6 (IL-6) is a multifunctional cytokine and high bloodstream levels of which have been associated with severe inflammatory diseases, such as dengue fever, sepsis, various cancers, and visceral leishmaniasis (VL). Rapid tests for the quantification of IL-6 would be of great assistance for the bedside diagnosis and treatment of diseases such as VL. We have developed a lateral flow assay (LFA) for rapid and colorimetric IL-6 detection, consisting of anti-IL-6 antibodies conjugated to gold nanoparticles (AuNPs). The optimal concentration of anti-IL-6 used in the conjugate was determined to be 800.0 μg/mL, based on an aggregation assay using LFA. A linear relationship between IL-6 standard concentration and color intensity was observed after 20 min, with a linear range between 1.25 ng/mL and 9,000 ng/mL. The limit of detection for this method was estimated at a t0.38 ng/mL. The concentration of IL-6 in five patients with severe VL was measured using LFA, and the results were consistent with those obtained using the cytometric bead array (CBA) method. A thorough analysis of the LFA membranes’ surface morphology, before and after sample contact, was performed using atomic force microscopy (AFM). The prototype described here is still being tested and improved, but this LFA will undoubtedly be of great help in the clinical quantification of IL-6.


Metabolites ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 240
Author(s):  
Alison Woodward ◽  
Alina Pandele ◽  
Salah Abdelrazig ◽  
Catherine A. Ortori ◽  
Iqbal Khan ◽  
...  

The integration of untargeted metabolomics and transcriptomics from the same population of cells or tissue enhances the confidence in the identified metabolic pathways and understanding of the enzyme–metabolite relationship. Here, we optimised a simultaneous extraction method of metabolites/lipids and RNA from ependymoma cells (BXD-1425). Relative to established RNA (mirVana kit) or metabolite (sequential solvent addition and shaking) single extraction methods, four dual-extraction techniques were evaluated and compared (methanol:water:chloroform ratios): cryomill/mirVana (1:1:2); cryomill-wash/Econospin (5:1:2); rotation/phenol-chloroform (9:10:1); Sequential/mirVana (1:1:3). All methods extracted the same metabolites, yet rotation/phenol-chloroform did not extract lipids. Cryomill/mirVana and sequential/mirVana recovered the highest amounts of RNA, at 70 and 68% of that recovered with mirVana kit alone. sequential/mirVana, involving RNA extraction from the interphase of our established sequential solvent addition and shaking metabolomics-lipidomics extraction method, was the most efficient approach overall. Sequential/mirVana was applied to study a) the biological effect caused by acute serum starvation in BXD-1425 cells and b) primary ependymoma tumour tissue. We found (a) 64 differentially abundant metabolites and 28 differentially expressed metabolic genes, discovering four gene-metabolite interactions, and (b) all metabolites and 62% lipids were above the limit of detection, and RNA yield was sufficient for transcriptomics, in just 10 mg of tissue.


Nanomaterials ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 768
Author(s):  
Hyung-Mo Kim ◽  
Chiwoo Oh ◽  
Jaehyun An ◽  
Seungki Baek ◽  
Sungje Bock ◽  
...  

Exosomes are attracting attention as new biomarkers for monitoring the diagnosis and prognosis of certain diseases. Colorimetric-based lateral-flow assays have been previously used to detect exosomes, but these have the disadvantage of a high limit of detection. Here, we introduce a new technique to improve exosome detection. In our approach, highly bright multi-quantum dots embedded in silica-encapsulated nanoparticles (M–QD–SNs), which have uniform size and are brighter than single quantum dots, were applied to the lateral flow immunoassay method to sensitively detect exosomes. Anti-CD63 antibodies were introduced on the surface of the M–QD–SNs, and a lateral flow immunoassay with the M–QD–SNs was conducted to detect human foreskin fibroblast (HFF) exosomes. Exosome samples included a wide range of concentrations from 100 to 1000 exosomes/µL, and the detection limit of our newly designed system was 117.94 exosome/μL, which was 11 times lower than the previously reported limits. Additionally, exosomes were selectively detected relative to the negative controls, liposomes, and newborn calf serum, confirming that this method prevented non-specific binding. Thus, our study demonstrates that highly sensitive and quantitative exosome detection can be conducted quickly and accurately by using lateral immunochromatographic analysis with M–QD–SNs.


2015 ◽  
Vol 864 ◽  
pp. 48-54 ◽  
Author(s):  
Eleni A. Sapountzi ◽  
Sotirios S. Tragoulias ◽  
Despina P. Kalogianni ◽  
Penelope C. Ioannou ◽  
Theodore K. Christopoulos

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
Xiaoshan Huang ◽  
Mingxin Guan ◽  
Zhuliangzi Lu ◽  
Yiping Hang

A new sensitive method for antimony (III) determination by graphite furnace atomic absorption spectrometry (GFAAS) has been developed by using N-benzoyl-N-phenylhydroxylamine (BPHA) and 1-butyl-3-methylimidazolium hexafluorophosphate ([C4mim][PF6]) single drop microextraction. The single drop microextraction (SDMM) system is more competitive compared with other traditional extraction methods. Under the optimized conditions, the limit of detection (signal-to-noise ratio is 3) and the enrichment factor of antimony (III) are 0.01 μg·L−1 and 112, respectively. The relative standard deviation of the 0.5 μg·L−1 antimony (III) is 4.2% (n=6). The proposed method is rather sensitive to determinate trace antimony (III) in water.


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