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Diagnostic Testing for Nickel Allergic Hypersensitivity: Patch Testing versus Lymphocyte Transformation Test
Published in Jurij J. Hostýnek, Howard I. Maibach, Nickel and the Skin, 2019
Jurij J. Hostýnek, Katherine E. Reagan, Howard I. Maibach
MELISA (memory lymphocyte immuno-stimulation assay), a lymphoproliferative assay, had been suggested by Stejskal et al. as a valuable instrument for the diagnosis of metal allergy (Stejskal et al., 1994). It is a modification of the LTT, specifically adapted to the study of lymphocyte reactivity to metals. Defibrinated blood is used in place of heparin, and monocyte and macrophage content is reduced by plastic adherence before analyzing for DNA synthesis, with the proviso that an increase in DNA synthesis be accompanied by the presence of lymphoblasts. Using the patch test as reference, sensitivity and specificity of the LTT and the MELISA were calculated, based on reactions from 34 patients with various metal contact allergies (Cederbrant et al., 1997). Neither sensitivity nor specificity was seen to differ significantly between the two in vitro test methods for the three metals gold, palladium, and nickel (se = 82%; sp = 17%). In both tests specificity was found to be unacceptably low for all three metals.
The Role of Dentistry in Cardiovascular Health and General Well-Being
Published in Stephen T. Sinatra, Mark C. Houston, Nutritional and Integrative Strategies in Cardiovascular Medicine, 2015
Most implants are titanium, and even though titanium is considered a very biocompatible material, approximately 4% of those tested with the MELISA® test are allergic. Memory lymphocyte immunostimulation assay (MELISA) is a blood test that detects Type-IV allergy to metals, chemicals, environmental toxins, and molds from one single blood sample.
Unveiling the underpinnings of various non-conventional ELISA variants: a review article
Published in Expert Review of Molecular Diagnostics, 2022
Rajesh Ahirwar, Akanksha Bhattacharya, Saroj Kumar
Another series of ELISA report the use of diverse forms of energies such as heat, microwave, pressure, and sound to enhance the binding interaction between an antigen and an antibody. Summarized as heat-mediated ELISA (HELISA), pressure-mediated ELISA (PELISA), sound-mediated ELISA (SELISA), and microwave-mediated ELISA (MELISA), these techniques reported assay time of as low as 5 minutes. A HELISA is performed with all incubations steps carried out at elevated temperature [40–50°C]; while PELISA report use of applied pressure of 1.5–2 × 105 Pa to achieve faster assay outcomes. Similarly, SELISA takes advantage of ultrasound (40 KHz in sonicator-water bath), and MELISA involves incubation of reaction mixtures in normal microwave oven (operating at power- 700 W) for varied time periods (40–100 sec) to achieve assay rapidity. Energies in the form of dry/wet heat, pressure, ultrasound waves, and microwave radiations contribute to enhance reaction rate in immunoassays by speeding the diffusion of reagents, and changing characteristics of antibodies/antigens in such a way allow stable interactions in comparatively lesser time. Also, the comparison of these energy-based ELISA for cross reactivity, sensitivity and error rate report at par analytical performance of these assays with conventional ELISA, suggesting that controlled energies do not lead to decreased activity of proteins/enzymes in ELISA [31–35].
Elevated Levels of Interleukins, Leukocyte Protein and Cathelicidin Antimicrobial Peptide are Strongly Associated with Early to Mid-Stage of Pythium insidiosum Infection in Rabbit Corneas
Published in Current Eye Research, 2022
Lalit Kishore Ahirwar, Savitri Sharma
The RT-qPCR result was further validated by mELISA. As very few cytokines were available which are specific to rabbit, only IL-8, IL-1β, MIP-1β and IL-17A expression were validated. Corneal tissue preserved at -80 °C were thawed on ice, washed with 1xPBS twice in petridish and 500 µL 1x lysis buffer [RIPA Buffer (10X), Cell Signaling Technology plus cOmplete™, EDTA-free Protease Inhibitor Cocktail, Roche (1 tablet in 10 ml of 1x RIPA buffer] was added and chopped using number #15 surgical sterile blade. The lysate was transferred into 1.5 ml eppendorf tube followed by 5 min incubation on ice and centrifugation (for 10 min at 14,000 × g, at 4 °C). Supernatant was collected in fresh tube without disturbing pellets and stored at −20 °C until use. Prior to performing mELISA for IL-1β, IL-8, IL-17A and Macrophage inflammatory protein (MIP)-1β using Quantibody® Rabbit Cytokine Array Q1(4), (RayBiotech protocol), protein concentrations were determined by BCA method [Bicinchoninic Acid (BCA) Protein Assay, G-Biosciences] and were adjusted to 1000 µg/mL with 1x RIPA buffer followed by two fold dilution with MiliQ-double autoclaved water. Microarray scanner G2565CA (Agilent Technologies) with Cy3 fluorescence detector was used to read all slides. Data was extracted using RayBiotech software and analyzed on Microsoft Excel spreadsheet.