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Gold Nanoparticles in Biology and Medicine
Published in Lev Dykman, Nikolai Khlebtsov, Gold Nanoparticles in Biomedical Applications, 2017
Membrane immunoassays (dot, slot and blot assays) commonly employ radioactive isotopes (125I, 14C, 3H) and enzymes (peroxydase, alkaline phosphatase, etc.) as labels. In 1984, four independent reports were published [172–175] in which CG was proposed for use as a label in solid-phase immunoassay. The use of GNP conjugates in solid-phase assay is based on the fact that the intense red coloration of a gold-containing marker allows the results of a reaction run on a solid carrier to be determined visually [176,177]. Immunogold methods in dot-blot assay outperform other techniques (e.g., enzyme immunoassay) in sensitivity (Table 2.1), rapidity, and low cost [178–180]. After an appropriate immunochemical reaction is run, the sizes of GNPs can be increased by enhancement with salts of silver [181] or gold (autometallography) [182], considerably increasing the method sensitivity. An optimized solid-phase assay using a densitometry system afforded a linear detection range from 1 pM to 1 μM [183], with detection limit of 100 aM, which was lowered to 100 zM by silver enhancement. The use of state-of-the-art instrumental detection methods, such as photothermal deflection of the laser beam, caused by heating of the local environment near absorbing particles by heating laser impulses [184], also ensures a very broad detection range (up to three orders of magnitude to the extent of several individual particles in a dot spot).
Infectious Bursal Disease
Published in Moayad N. Khalaf, Michael Olegovich Smirnov, Porteen Kannan, A. K. Haghi, Environmental Technology and Engineering Techniques, 2020
The dot blot assay is a good alternative to ELISA and IFAT in the serodiagnosis. Cruz-Coy et al. (1993) used monoclonal antibody (mAb) developed against a variant subtype of IBDV, to recognize all six serologic subtypes of IBDV and three untyped IBDV by dot blot method. Anil et al. (2002) reported that dot blot was as equally sensitive as RT-PCR.
Production of recombinant lethal factor of Bacillus anthracis in Bacillus subtilis
Published in Preparative Biochemistry & Biotechnology, 2021
Mahboobeh Gholami, Majid Moghbeli, Farshid Kafilzadeh, Mohammad Kargar, Mariam Bikhof Torbati, Ashkan Tavizi, Sally Bellevile, Javad Hatami, Zahra Eslami
To confirm the expression of the lethal factor protein, dot-blot immunoassay method and ELISA test were carried out. In dot-blot immunoassay, the recombinant protein was spotted onto a membrane and was hybridized with an antibody probe. As it is shown in Fig. 3, the expression of the LF protein was verified as the color altered in the presence of the primary and the secondary antibody. ELISA tests also re-confirmed the expression of this protein.