scholarly journals Fluorimetric Immunoassay for Multianalysis of Aflatoxins

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Lizy Kanungo ◽  
Sunil Bhand
Keyword(s):  
High Sensitivity ◽  
High Specificity ◽  
Standard Addition ◽  
Cross Reactivity ◽  
Addition Method ◽  
Milk Samples ◽  
Capture Antibody ◽  
Multianalyte Detection ◽  
Microwell Plate ◽  
Detection And Quantification

A sensitive fluorimetric ELISA was developed for the analysis of aflatoxins. The assay was performed in a 384 microwell plate, wherein high specificity monoclonal antibody against AFM1 (mAb-AFM1) was used as capture antibody and FITC conjugated secondary antibody was used for detection and quantification of the analyte. The linear range of the immunoassay was found to be 6.25–50 pg/mL. AFM1 as low as 1 pg/mL was detected by this method with assay volume 40 μL. The multi-analysis of different aflatoxins was also investigated in the microwell plate, based on the cross-reactivity (CR) approach. Real milk samples were tested along with certified reference material by standard addition method and recovery analysis was done. The mAb-AFM1 showed 23.2% CR with AFB1, 50% CR with respect to AFM2, and least CR towards AFG1 (<1%). Furthermore, mixture analysis of AFM2 and AFB1 was carried out at specific concentrations of AFM1. The advantages of this developed immunoassay are high sensitivity, high throughput, multianalyte detection, versatility, and ease of handling.

scholarly journals A 16S rDNA-based nested PCR protocol to detect Campylobacter gracilis in oral infections

2003 ◽  
Vol 17 (2) ◽  
pp. 142-146 ◽  
Author(s):  
José Freitas Siqueira Júnior ◽  
Isabela das Neves Rôças
Keyword(s):  
16S Rdna ◽  
High Sensitivity ◽  
High Specificity ◽  
Cross Reactivity ◽  
Nested Polymerase Chain Reaction ◽  
Oral Infections ◽  
Root Canals ◽  
Infected Root ◽  
Pcr Products ◽  
Species Specific

The aim of this study was to describe a 16S rDNA-based nested polymerase chain reaction (nPCR) assay to investigate the occurrence of Campylobacter gracilis in oral infections. Samples were collected from ten infected root canals, ten cases of acute periradicular abscesses and eight cases of adult marginal periodontitis. DNA extracted from the samples was initially amplified using universal 16S rDNA primers. A second round of amplification used the first PCR products to detect C. gracilis using oligonucleotide primers designed from species-specific 16S rDNA signature sequences. The nPCR assay used in this study showed a detection limit of 10 C. gracilis cells and no cross-reactivity was observed with nontarget bacteria. C. gracilis was detected in the three types of oral infections investigated - 4/10 infected root canals; 2/10 acute periradicular abscesses; and 1/8 subgingival specimens from adult periodontitis. The method proposed in this study showed both high sensitivity and high specificity to directly detect C. gracilis in samples from root canal infections, abscesses, and subgingival plaque. Our findings confirmed that C. gracilis may be a member of the microbiota associated with distinct oral infections, and its specific role in such diseases requires further clarification.

scholarly journals Performance of recombinant proteins in diagnosis and differentiation of canine visceral leishmaniasis infected and vaccinated dogs

2020 ◽  
Vol 10 (3) ◽  
pp. 165-171
Author(s):  
Ingrid E. Pereira ◽  
Kyssia P. Silva ◽  
Laura M. Menegati ◽  
Aimara C. Pinheiro ◽  
Elaine A. O. Assunção ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Recombinant Proteins ◽  
Serological Test ◽  
High Sensitivity ◽  
High Specificity ◽  
Cross Reactivity ◽  
Reservoir Host ◽  
Superior Performance ◽  
Canine Visceral Leishmaniasis ◽  
Roc Curve Analysis

AbstractControl of canine visceral leishmaniasis (CVL), a major zoonotic disease in Brazil and many other tropical and subtropical countries, remains difficult as an accurate and reliable diagnosis is still missing. In endemic regions, infected dogs are the main parasitic reservoir host of human Visceral leishmaniasis (VL) infection. Vaccination of dogs against Leishmania infection constitutes an important strategy to prevent or to better control CVL, thus, a serological test that can discriminate between antibodies induced by immunization versus infection is highly desirable in order to improve and simplify diagnosis. Here, four recombinant proteins were evaluated for their ability to detect and differentiate between dogs that are infected with Leishmania or have been immunized with the anti-Leishmania vaccine Leish-Tec®. Receiver operating characteristic (ROC) curve analysis of the four Leishmania-specific IgG ELISA revealed superior performance of rK28, followed by rKLO8, rK39 and rLb6H. The rK28-based ELISA revealed not only the best accuracy against CVL, but also the lowest cross-reactivity with sera from Leish-Tec® immunized dogs. Our data show that the rK28-based ELISA is highly suitable for CVL screening as it shows high sensitivity with simultaneous low cross-reactivity. Further, the high specificity of the rKLO8 indicates its suitability for the confirmation of CVL diagnosis.

Spectrophotometric Method for the Assay of Chlorhexiidine Acetate

2014 ◽  
Vol 912-914 ◽  
pp. 321-324
Author(s):  
Yan Hong Lei
Keyword(s):  
Spectrophotometric Method ◽  
High Sensitivity ◽  
Standard Addition ◽  
Standard Addition Method ◽  
Addition Method ◽  
Ultraviolet Spectrophotometry ◽  
Good Linear ◽  
Sensitivity And Selectivity

To establish the determination of the content of chlorhexiidine acetate by ultraviolet spectrophotometry .The spectra (range: λ=200~320 nm) were processed. The linear ranges of chlorhexidine acetate was at 2~ 20 μg mL-1 .the chlorhexiidine acetate showed good linear correlation.The ultraviolet spectrophotometry data of the samples were also used to evaluate the samples quantitative composition.It was shown that recovery of the method by standard addition method was respectively valued 99.55% for chlorhexiidine acetate .The method was simple,reliable,accurate,reproducible with high sensitivity and selectivity.

Spectrophotometric Method for the Assay of Aspirin

2014 ◽  
Vol 602-605 ◽  
pp. 2097-2100
Author(s):  
Yan Hong Lei
Keyword(s):  
Spectrophotometric Method ◽  
High Sensitivity ◽  
Standard Addition ◽  
Standard Addition Method ◽  
Addition Method ◽  
Ultraviolet Spectrophotometry ◽  
Selective Method ◽  
Good Linear ◽  
Sensitivity And Selectivity

A selective method was developed for the determination of aspirin by Spectrophotometric method .The spectra (range: λ=200~320 nm) were processed. The linear ranges of aspirin was at 50.28 ~ 125.7μg • mL-1 .the aspirin showed good linear correlation.The ultraviolet spectrophotometry data of the samples were also used to evaluate the samples’ quantitative composition.It was shown that recovery of the method by standard addition method was respectively valued 102.06% for aspirin .The method was simple,reliable,accurate,reproducible with high sensitivity and selectivity.

scholarly journals Performance Characteristics of Four High-Throughput Immunoassays for Detection of IgG Antibodies against SARS-CoV-2

2020 ◽  
Vol 58 (8) ◽  
Author(s):  
Elitza S. Theel ◽  
Julie Harring ◽  
Heather Hilgart ◽  
Dane Granger
Keyword(s):  
High Throughput ◽  
High Sensitivity ◽  
High Specificity ◽  
Cross Reactivity ◽  
Clinical Diagnostics ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Predictive Values ◽  
Serologic Tests ◽  
Convalescent Phase

ABSTRACT The role of serologic testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in both the clinical and public health settings, will continue to evolve as we gain increasing insight into our immune response to the virus. Here, we evaluated four high-throughput serologic tests for detection of anti-SARS-CoV-2 IgG antibodies, from Abbott Laboratories (Abbott Park, IL), Epitope Diagnostics, Inc. (San Diego, CA), Euroimmun (Lubeck, Germany), and Ortho-Clinical Diagnostics (Rochester, NY), using a panel of serially collected serum samples (n = 224) from 56 patients with confirmed coronavirus disease 2019 (COVID-19), healthy donor sera from 2018, and a cross-reactivity serum panel collected in early 2020. The sensitivities of the Abbott, Epitope, Euroimmun, and Ortho-Clinical IgG assays in convalescent-phase serum samples collected more than 14 days post-symptom onset or post-initial positive reverse transcriptase PCR (RT-PCR) result were 92.9% (78/84), 88.1% (74/84), 97.6% (82/84), and 98.8% (83/84), respectively. Among unique convalescent patients, sensitivities of the Abbott, Epitope, Euroimmun, and Ortho-Clinical anti-SARS-CoV-2 IgG assays were 97.3% (36/37), 73% (27/37), 94.6% (35/37), and 97.3% (36/37), respectively. Overall assay specificity/positive predictive values based on a 5% prevalence rate were 99.6%/92.8%, 99.6%/90.6%, 98.0%/71.2%, and 99.6%/92.5%, respectively, for the Abbott, Epitope, Euroimmun, and Ortho-Clinical IgG assays. In conclusion, we show high sensitivity in convalescent-phase sera and high specificity for the Abbott, Euroimmun, and Ortho-Clinical anti-SARS-CoV-2 IgG assays. With the unprecedented influx of commercially available serologic tests for detection of antibodies against SARS-CoV-2, it remains imperative that laboratories thoroughly evaluate such assays for accuracy prior to implementation.

Spectrophotometric Method for the Assay of Norfloxacin Lactate

2014 ◽  
Vol 989-994 ◽  
pp. 993-996
Author(s):  
Yan Hong Lei
Keyword(s):  
Spectrophotometric Method ◽  
High Sensitivity ◽  
Standard Addition ◽  
Standard Addition Method ◽  
Addition Method ◽  
Selective Method

A selective method was developed for the determination of norfloxacin lactate by Spectrophotometric method. The method was simple with high sensitivity and selectivity.It was shown that recovery of the method by standard addition method was respectively valued 102.05% for norfloxacin lactate

Spectrophotometric Method in the Determination of Aminophylline Content

2014 ◽  
Vol 602-605 ◽  
pp. 2395-2398
Author(s):  
Yan Hong Lei
Keyword(s):  
Spectrophotometric Method ◽  
High Sensitivity ◽  
Standard Addition ◽  
Standard Addition Method ◽  
Addition Method ◽  
Selective Method

A selective method was developed for the determination of aminophylline by Spectrophotometric method. The method was simple with high sensitivity and selectivity.It was shown that recovery of the method by standard addition method was respectively valued 99.21% for aminophylline.

scholarly journals Assessing the risk of heavy metals contamination in milk from Pakistan

2021 ◽  
Vol 3 (1) ◽  
pp. 107-113
Author(s):  
ZH Shar ◽  
OP Pirhot ◽  
HH Shar ◽  
MK Channa
Keyword(s):  
Toxic Metals ◽  
Standard Addition ◽  
Essential Elements ◽  
Essential Metals ◽  
Addition Method ◽  
Milk Samples ◽  
Fresh Milk ◽  
Heavy Metals Contamination ◽  
External Standard ◽  
Cadmium And Lead

Milk is an essential component of human food, and natural source of many important elements. Besides essential elements it is also became a source of toxic metals due to heavy environmental pollution. In order to assess the essential metals (calcium) and toxic metals (cadmium and lead) in milk, sixty different fresh milk samples were analyzed by using atomic absorption spectroscopy (AAS). Accuracy and precession were checked by external standard addition method. Calcium was found in all samples with highest concentration (24µg/L) in camel milk. Lead was found in all milk samples with mean level 3.14µg/L and highest concentration found in packed milk sample 6.7µg/L. Cadmium was detected in 33% of total samples analyzed with range of (1.1-3.1µg/L). Results of this study will be helpful in setting the standards in one of the most consumed commodities in Pakistan.

scholarly journals Radiochemistry, Production Processes, Labeling Methods, and ImmunoPET Imaging Pharmaceuticals of Iodine-124

Molecules ◽  
2021 ◽  
Vol 26 (2) ◽  
pp. 414
Author(s):  
Krishan Kumar ◽  
Arijit Ghosh
Keyword(s):  
Physical Properties ◽  
Imaging Modality ◽  
Target Selection ◽  
Positron Emission Tomography Imaging ◽  
High Sensitivity ◽  
High Specificity ◽  
Production Processes ◽  
Positron Emission ◽  
Significant Research ◽  
Clinical Environments

Target-specific biomolecules, monoclonal antibodies (mAb), proteins, and protein fragments are known to have high specificity and affinity for receptors associated with tumors and other pathological conditions. However, the large biomolecules have relatively intermediate to long circulation half-lives (>day) and tumor localization times. Combining superior target specificity of mAbs and high sensitivity and resolution of the PET (Positron Emission Tomography) imaging technique has created a paradigm-shifting imaging modality, ImmunoPET. In addition to metallic PET radionuclides, 124I is an attractive radionuclide for radiolabeling of mAbs as potential immunoPET imaging pharmaceuticals due to its physical properties (decay characteristics and half-life), easy and routine production by cyclotrons, and well-established methodologies for radioiodination. The objective of this report is to provide a comprehensive review of the physical properties of iodine and iodine radionuclides, production processes of 124I, various 124I-labeling methodologies for large biomolecules, mAbs, and the development of 124I-labeled immunoPET imaging pharmaceuticals for various cancer targets in preclinical and clinical environments. A summary of several production processes, including 123Te(d,n)124I, 124Te(d,2n)124I, 121Sb(α,n)124I, 123Sb(α,3n)124I, 123Sb(3He,2n)124I, natSb(α, xn)124I, natSb(3He,n)124I reactions, a detailed overview of the 124Te(p,n)124I reaction (including target selection, preparation, processing, and recovery of 124I), and a fully automated process that can be scaled up for GMP (Good Manufacturing Practices) production of large quantities of 124I is provided. Direct, using inorganic and organic oxidizing agents and enzyme catalysis, and indirect, using prosthetic groups, 124I-labeling techniques have been discussed. Significant research has been conducted, in more than the last two decades, in the development of 124I-labeled immunoPET imaging pharmaceuticals for target-specific cancer detection. Details of preclinical and clinical evaluations of the potential 124I-labeled immunoPET imaging pharmaceuticals are described here.

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