Antibody microarray immunoassay for screening and differential diagnosis of upper respiratory tract viral pathogens.

Autor: Plotnikova MA; Smorodintsev Research Institute of Influenza, St. Petersburg, Russia. Electronic address: biomalinka@gmail.com., Klotchenko SA; Smorodintsev Research Institute of Influenza, St. Petersburg, Russia., Lebedev KI; Smorodintsev Research Institute of Influenza, St. Petersburg, Russia; Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia., Lozhkov AA; Smorodintsev Research Institute of Influenza, St. Petersburg, Russia; Peter the Great St. Petersburg Polytechnic University, St. Petersburg, Russia., Taraskin AS; Smorodintsev Research Institute of Influenza, St. Petersburg, Russia; Peter the Great St. Petersburg Polytechnic University, St. Petersburg, Russia., Gyulikhandanova NE; Smorodintsev Research Institute of Influenza, St. Petersburg, Russia; Peter the Great St. Petersburg Polytechnic University, St. Petersburg, Russia., Ramsay ES; Smorodintsev Research Institute of Influenza, St. Petersburg, Russia., Vasin AV; Smorodintsev Research Institute of Influenza, St. Petersburg, Russia; Peter the Great St. Petersburg Polytechnic University, St. Petersburg, Russia.
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 2020 Mar; Vol. 478, pp. 112712. Date of Electronic Publication: 2019 Nov 27.
DOI: 10.1016/j.jim.2019.112712
Abstrakt: Upper respiratory tract infections are the world's most common infectious disease. The etiologic agents behind upper respiratory tract infections (URTIs) are, in fact, a diverse set of pathogens such as influenza, parainfluenza, adenovirus, rhinovirus, and others. More than 200 pathogens are known to be involved. Differential diagnosis of viral infections is sometimes complicated by their diversity or similarity of clinical presentation. This work is devoted to the development of a method which enables simultaneous detection of six common viral URTI pathogens: IAV; IBV; RSV; hAdV; hPIV2; and hPIV3. Antibody microarray technology is utilized to accomplish the analysis. In preparation for protein microchip creation, we produced, characterized, and selected approximately 50 monoclonal antibodies; for each of the aforementioned pathogens, an optimal monoclonal antibody pair was selected. A protein microchip was created, and its core working conditions were optimized. With a balance between convenience and maximal assay sensitivity in mind, a one-step analysis approach was developed for accomplishing the ELISA-like "sandwich" interaction on the manufactured microchip (antibody microarray). Reference viral strains were used to establish the lower limits of detection (LoD) for the assay. For IAV, the LoD was 0.25 ng/ml total viral protein. For other viruses, the LoD ranged from 1 to 2 ng/ml total protein. These sensitivity limits are slightly better than those of standard ELISA, but inferior to those of PCR. Overall, we believe that the developed microchip is a good alternative to existing methods, allowing relatively quick (overnight), inexpensive, simultaneous screening of several pathogens. The design of the antibody microarray is conducive to further development, and the panel of analyzed pathogens can be expanded to include approximately 50 members.
Competing Interests: Declaration of Competing Interest The authors declare that there are no conflicts of interest.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: