scholarly journals A Novel, Inexpensive In-House Immunochromatographic Strip Test for Cryptococcosis Based on the Cryptococcal Glucuronoxylomannan Specific Monoclonal Antibody 18B7

Diagnostics ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 758
Author(s):  
Pakornswit Sathongdejwisit ◽  
Kritsada Pruksaphon ◽  
Akarin Intaramat ◽  
Pisinee Aiumurai ◽  
Nitat Sookrung ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Lower Limit ◽  
Limit Of Detection ◽  
Latex Agglutination ◽  
Specific Monoclonal Antibody ◽  
Serum Samples ◽  
Immunochromatographic Strip ◽  
Strip Test ◽  
Sensitivity Specificity ◽  
Capsular Antigens

The aim of this study was to develop a novel lateral flow immunochromatoghaphic strip test (ICT) for detecting cryptococcal polysaccharide capsular antigens using only a single specific monoclonal antibody, mAb 18B7. The mAb 18B7 is a well characterized antibody that specifically binds repeating epitopes displayed on the cryptococcal polysaccharide glucuronoxylomannan (GXM). We validated the immunoreactivities of mAb 18B7 against capsular antigens of different cryptococcal serotypes. The mAb 18B7 ICT was constructed as a sandwich ICT strip and the antibody serving in the mobile phase (colloidal gold conjugated mAb 18B7) to bind one of the GXM epitopes while the stationary phase antibody (immobilized mAb18B7 on test line) binding to other remaining unoccupied epitopes to generate a positive visual readout. The lower limit of detection of capsular antigens for each of the Cryptococcus serotypes tested was 0.63 ng/mL. No cross-reaction was found against a panel of antigens isolated from cultures of other pathogenic fungal, except the crude antigen of Trichosporon sp. with the lower limit of detection of 500 ng/mL (~800 times higher than that for cryptococcal GXM). The performance of the mAb 18B7 ICT strip was studied using cerebrospinal fluid (CSF) and serum and compared to commercial diagnostic kits (latex agglutination CALAS and CrAg IMMY). The sensitivity, specificity and accuracy of the mAb18B7 ICT with CSF from patients with confirmed cryptococcal meningitis were 92.86%, 100% and 96.23%, respectively. No false positives were observed with samples from non-cryptococcosis patients. With serum samples, the mAb 18B7 ICT gave a sensitivity, specificity and accuracy of 96.15%, 97.78% and 96.91%, respectively. Our results show that the mAb 18B7 based ICT was reliable, reproducible, and cost-effective as a point-of-care immunodiagnostic test for cryptococcosis. The mAb 18B7 ICT may be particularly useful in countries where commercial kits are not available or affordable.

scholarly journals Use of Protein-Specific Monoclonal Antibody-Based Latex Agglutination for Rapid Diagnosis of Burkholderia pseudomallei Infection in Patients with Community-Acquired Septicemia

2007 ◽  
Vol 14 (6) ◽  
pp. 811-812 ◽  
Author(s):  
Pattama Ekpo ◽  
Utane Rungpanich ◽  
Supinya Pongsunk ◽  
Pimjai Naigowit ◽  
Vimon Petkanchanapong
Keyword(s):  
Monoclonal Antibody ◽  
Blood Culture ◽  
Burkholderia Pseudomallei ◽  
Latex Agglutination ◽  
Rapid Diagnosis ◽  
Agglutination Test ◽  
Latex Agglutination Test ◽  
Specific Monoclonal Antibody ◽  
Predictive Values ◽  
Sensitivity Specificity

ABSTRACT A latex agglutination test employing monoclonal antibody specific to a 30-kDa protein of Burkholderia pseudomallei was used to detect the organisms in blood culture specimens from 1,139 patients with community-acquired septicemia. The sensitivity, specificity, and positive and negative predictive values of the test were 96.75%, 99.61%, 96.75%, and 99.61%, respectively.

Iron Oxide Nanoparticles Conjugated Monoclonal Antibody for Immunochromatographic Strip Test

2011 ◽  
Vol 364 ◽  
pp. 30-34
Author(s):  
Mohamad Nor Noorhashimah ◽  
Dyana Zakaria Nor ◽  
Azlan Abdul Aziz ◽  
Rahmah Noordin ◽  
Abdul Razak Khairunisak
Keyword(s):  
Monoclonal Antibody ◽  
Iron Oxide ◽  
Iron Oxide Nanoparticles ◽  
Precipitation Method ◽  
Oxide Nanoparticles ◽  
Serum Samples ◽  
Immunochromatographic Strip ◽  
Transmission Electron ◽  
Strip Test ◽  
Icg Strip

In this study, the synthesis of iron oxide nanoparticles (IONPs) and immunochromatographic (ICG) strip test of iron oxide conjugated with monoclonal antibody (IONPs-Ab) have been developed. The IONPs were synthesised using precipitation method and dispersed in water by applying polyethylene glycol coating. Several parameters that affected conjugations of the IONPs-Ab were studied namely iron oxide concentration, antibody volume, stabilizer concentration and the amount of SiPEG coating. From the transmission electron microscopy (TEM) image, the size of IONPs obtained was ~14 nm. Conjugate was tested with the ICG strip test lined with the antigen. The results showed 1.0μl SiPEG coating, 1 M IONPs concentration, 20μl of 1mg/ml antibody volume and 1% bovine serum albumin (BSA) were the optimum. Moreover, the IONPs-Ab was also tested with the Brugian filariasis positive and negative serum samples. The results showed positive result for the patient serum and negative result for the non-patient serum in just 15 minutes.

scholarly journals Immunochromatographic Strip Test for Rapid Detection of Nevirapine in Plasma Samples from Human Immunodeficiency Virus-Infected Patients

2007 ◽  
Vol 51 (9) ◽  
pp. 3361-3363 ◽  
Author(s):  
Tim R. Cressey ◽  
Sawitree Nangola ◽  
Yardpiroon Tawon ◽  
Mookda Pattarawarapan ◽  
Marc Lallemant ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Limit Of Detection ◽  
Drug Level ◽  
Plasma Samples ◽  
Immunochromatographic Strip ◽  
Resource Limited ◽  
Qualitative Detection ◽  
Immunodeficiency Virus ◽  
Immunochromatographic Strip Test ◽  
Strip Test

ABSTRACT We report a novel one-step immunochromatographic strip test for the rapid, qualitative detection of nevirapine in plasma samples from human immunodeficiency virus-infected patients. The sensitivity was 100% (95% confidence interval [95% CI], 97.8 to 100%), and the specificity was 99.5% (95% CI, 97.2 to 99.9%). The limit of detection was 25 ng/ml. Immunochromatographic strip tests are simple, rapid, and cheap assays that could greatly facilitate drug level monitoring in resource-limited settings.

scholarly journals Rapid immunoassays for detection of anabolic nortestosterone in dietary supplements

2013 ◽  
Vol 31 (No. 5) ◽  
pp. 514-519 ◽  
Author(s):  
B. Holubová ◽  
S. Göselová ◽  
L. Ševčíková ◽  
M. Vlach ◽  
M. Blažková ◽  
...  
Keyword(s):  
Dietary Supplements ◽  
Rapid Detection ◽  
Visual Detection ◽  
Limit Of Detection ◽  
Polyclonal Antibodies ◽  
Cross Reactivity ◽  
Rapid Analysis ◽  
Experimental Conditions ◽  
Immunochromatographic Strip ◽  
Strip Test

An enzyme immunoassay (ELISA) and an immunochromatographic strip were designed for a rapid detection of nortestosterone in dietary supplements. Two polyclonal antibodies and two types of nortestosterone-protein coating conjugates were tested to develop the most appropriate method. Under optimal experimental conditions, the most sensitive ELISA achieved the IC<sub>50 </sub>and the limit of detection values of 6.41 and 0.09 ng/ml, respectively. The assay specificity was tested measuring cross-reactivity of several steroids. The interference with the assay was negligible (&lt; 0.1%), except for cross-reactivity with another frequently abused steroid testosterone (23%). The optimised gold particle-based immunochromatographic strip provided in semi-quantitative test a visual detection limit of 1&nbsp;ng/ml. None of these methods showed the interference using a filtrate of the suspension of non-contaminated sample. After the validation for particular matrices, the ELISA and the strip test could be useful tools for a rapid analysis of nortestosterone in crude extracts of dietary supplements.

Monoclonal antibodies evaluated for use in a screening assay for conjugates of human serum albumin and pyridoxal 5'-phosphate.

1989 ◽  
Vol 35 (8) ◽  
pp. 1756-1760 ◽  
Author(s):  
B B Miller ◽  
W E Turner
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Human Serum Albumin ◽  
Human Serum ◽  
Limit Of Detection ◽  
Screening Method ◽  
Serum Samples ◽  
Screening Assay ◽  
Antibody Avidity ◽  
Negative Results

Abstract This enzyme immunoassay (EIA) was developed to measure pyridoxal 5'-phosphate bound to albumin (PLP-HSA) in human serum. The monoclonal antibody titer was 1:2000 and a sequential saturation analysis curve, prepared with samples containing from 10 to 1000 nmol/L, showed a 50% inhibition of antibody at 50 nmol of the conjugate per liter. The lower limit of detection for PLP-HSA was 10 nmol/L, a sensitivity 1000-fold greater than that for any potential interferent. When serum samples gave negative results in the assay, we compared the antigenicity of the principal sites for PLP binding on HSA. It was apparent that the preferred physiological site was not antigenic; however, three additional sites for PLP binding on HSA elicited comparable antibody avidity. This EIA is potentially quite sensitive and specific for PLP-HSA, but considerable additional effort is required to convert serum PLP to an HSA-bound form detectable in the assay, which limits its application as a screening method.

scholarly journals Development of a Sensitive Bead-Based Assay for Enhanced Monoclonal Antibody Detection

2011 ◽  
Vol 1346 ◽  
Author(s):  
Manuel E. Ruidíaz ◽  
Natalie Mendez ◽  
Ana B. Sanchez ◽  
Bradley T. Messmer ◽  
Andrew C. Kummel
Keyword(s):  
Monoclonal Antibody ◽  
Limit Of Detection ◽  
Solid Phase ◽  
Solid Phase Peptide Synthesis ◽  
Fluorescent Labeling ◽  
Antibody Detection ◽  
Serum Samples ◽  
Mimetic Peptide ◽  
Mimetic Peptides ◽  
Zero Response

ABSTRACTMonoclonal antibodies are increasingly used in the treatment of cancer due to their enhanced targeting and immune system stimulation properties. Dosage guidelines typically do not account for personal cancer load or metabolism, thereby possibly affecting treatment outcome or causing unwanted side effects. The requirement for an assay that can quickly and precisely measure the concentration of the monoclonal antibody in a serum sample of a patient during therapy is unmet. A bead-based assay with peptide antigen mimetics has been developed to rapidly determine the concentration of antibody drug present in serum specimens with high sensitivity. Alemtuzumab (anti-CD52) and rituximab (anti-CD20) antigen mimetic peptides, as discovered by phage display, were synthesized on 10 um TentaGel resin beads using conventional solid phase peptide synthesis techniques. The beads were modified to allow for multiplexing and microfluidic handling via fluorescent labeling and magnetic functionalization. The antigen-displaying fluoromagnetic particles were incubated with spiked serum samples which allowed free antibody to be captured. Primary antibody detection was performed on alemtuzumab while rituximab detection was used to compensate for non-specific serum binding to the beads. After washing, the beads were incubated with a fluorescently tagged secondary label for detection by flow cytometry. (Results) A fast, low cost, specific assay has been developed with several key techniques which allows detection at low concentration (0.1ug/ml) of spiked samples. Primary to achieving this detection limit was the implementation of a compensation scheme where two antigen mimetic peptides behave linearly (R2=0.996) which enables the calculation of the zero response of the antigen mimetic peptide of interest (alemtuzumab antigen mimetic) while measuring the zero response of the compensatory antigen mimetic peptide (rituximab antigen mimetic) during primary assay measurement. This reduces fluorescence response variation due to variations present due to sample preparation, storage and different patients because of the equivalent interactions these effects have on the compensatory beads. The developed assay is therefore robust against serum variation and enables a lower limit of detection.

scholarly journals Development of Aflatoxin B1-Lysine Adduct Monoclonal Antibody for Human Exposure Studies

2001 ◽  
Vol 67 (6) ◽  
pp. 2712-2717 ◽  
Author(s):  
Jia-Sheng Wang ◽  
Salahaddin Abubaker ◽  
Xia He ◽  
Guiju Sun ◽  
Paul T. Strickland ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
High Risk ◽  
Bovine Serum Albumin ◽  
Dna Adducts ◽  
Human Exposure ◽  
Limit Of Detection ◽  
Serum Samples ◽  
Bovine Serum Albumin Conjugate ◽  
Aflatoxin B

ABSTRACT Mouse monoclonal antibodies were developed against a synthetic aflatoxin B1 (AFB)-lysine–cationized bovine serum albumin conjugate. The isotype of one of these antibodies, IIA4B3, has been classified as immunoglobulin G1(λ). The affinity and specificity of IIA4B3 were further characterized by a competitive radioimmunoassay. The affinities of IIA4B3 for AFB and its associated adducts and metabolites are ranked as follows: AFB-lysine > 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-AFB > AFB = 8,9-dihydro-8-(N 7-guanyl)-9-hydroxy-AFB > aflatoxin M1 > aflatoxin Q1. IIA4B3 had about a 10-fold higher affinity for binding to AFB-lysine adduct than to AFB when 3H-AFB–lysine was used as the tracer. The concentration for 50% inhibition for AFB-lysine was 0.610 pmol; that for AFB was 6.85 pmol. IIA4B3 had affinities at least sevenfold and twofold higher than those of 2B11, a previously developed antibody against parent AFB, for the major aflatoxin-DNA adducts 8,9-dihydro-8-(N 7-guanyl)-9-hydroxy-AFB and 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-AFB, respectively. An analytical method based on a competitive radioimmunoassay with IIA4B3 and3H-AFB–lysine was validated with a limit of detection of 10 fmol of AFB-lysine adduct. The method has been applied to the measurement of AFB-albumin adduct levels in human serum samples collected from the residents of areas at high risk for liver cancer.

Development of a monoclonal antibody for the detection of xylazine in milk and its use in an immunochromatographic strip

2021 ◽  
Author(s):  
Mengjia Chao ◽  
Liqiang Liu ◽  
Aihong Wu ◽  
Shanshan Song ◽  
Xinxin Xu ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Gold Nanoparticle ◽  
Limit Of Detection ◽  
Lateral Flow ◽  
Immunochromatographic Strip ◽  
Naked Eye ◽  
Milk Samples ◽  
Flow Test ◽  
Lateral Flow Test

A gold nanoparticle-based lateral-flow test (GNT) strip was developed to detect xylazine (XYL) in milk. And the limit of detection (LOD) and cut-off value of the GNT assay were evaluated to be 20 and 200 ng mL−1 in milk samples by the naked eye.

Immunodiagnosis of paramphistomosis using monoclonal antibody-based sandwich ELISA for detection ofParamphistomum gracilecirculating 16 kDa antigen

Parasitology ◽  
2017 ◽  
Vol 144 (7) ◽  
pp. 899-903 ◽  
Author(s):  
PANAT ANURACPREEDA ◽  
KULLANID TEPSUPORNKUL ◽  
RUNGLAWAN CHAWENGKIRTTIKUL
Keyword(s):  
Monoclonal Antibody ◽  
Detection Method ◽  
Sandwich Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Microtitre Plate ◽  
Serum Samples ◽  
Fecal Samples ◽  
Lower Detection Limit ◽  
Lower Detection ◽  
Sensitivity Specificity

SUMMARYIn this study, we have produced a monoclonal antibody (MoAb) against 16 kDa antigen ofParamphistomum gracile(16 kDaAgPg), and developed an accurate sandwich enzyme-linked immunosorbent assay (sandwich ELISA) for the detection of circulating 16 kDaAg in the serum and fecal samples from cattle naturally infected withP. gracile. MoAb 1D10 was immobilized on a microtitre plate, and the antigen in the samples was captured and detected with biotinylated rabbit anti-16 kDaAgPg antibody. The lower detection limit of sandwich ELISA was 3·5 pg mL−1, and no cross-reaction with other parasite antigens was evaluated. The reliability of the assay was examined using the serum and fecal samples from cattle naturally infected withP. gracile, Fasciola gigantica, Moniezia benedeni, Trichurissp.,Strongyloidessp., strongylids and non-infected animals. The sandwich ELISA showed the sensitivity, specificity and accuracy at 98·33, 100 and 99·55% (serum samples), and 96·67, 100 and 99·09% (fecal samples). Therefore, this detection method is a rapid and excellent potential assay for the accurate diagnosis of paramphistomosis.

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