tetano
Editor, Senior Moderator
J Immunol Methods. 2019 Nov 26:112712. doi: 10.1016/j.jim.2019.112712. [Epub ahead of print] [h=1]Antibody microarray immunoassay for screening and differential diagnosis of upper respiratory tract viral pathogens.[/h]
Plotnikova MA[SUP]1[/SUP], Klotchenko SA[SUP]2[/SUP], Lebedev KI[SUP]3[/SUP], Lozhkov AA[SUP]4[/SUP], Taraskin AS[SUP]4[/SUP], Gyulikhandanova NE[SUP]4[/SUP], Ramsay ES[SUP]2[/SUP], Vasin AV[SUP]4[/SUP].
[h=3]Author information[/h] 1 Smorodintsev Research Institute of Influenza, St. Petersburg, Russia. Electronic address: biomalinka@gmail.com. 2 Smorodintsev Research Institute of Influenza, St. Petersburg, Russia. 3 Smorodintsev Research Institute of Influenza, St. Petersburg, Russia; Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia. 4 Smorodintsev Research Institute of Influenza, St. Petersburg, Russia; Peter the Great St. Petersburg Polytechnic University, St. Petersburg, Russia.
[h=3]Abstract[/h] 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.
Copyright ? 2019. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Antibody array; Diagnosis of viral diseases; Monoclonal antibodies; Multiplex immunoassay; Multiplex molecular diagnostics; Severe acute respiratory infections
PMID: 31783022 DOI: 10.1016/j.jim.2019.112712
Plotnikova MA[SUP]1[/SUP], Klotchenko SA[SUP]2[/SUP], Lebedev KI[SUP]3[/SUP], Lozhkov AA[SUP]4[/SUP], Taraskin AS[SUP]4[/SUP], Gyulikhandanova NE[SUP]4[/SUP], Ramsay ES[SUP]2[/SUP], Vasin AV[SUP]4[/SUP].
[h=3]Author information[/h] 1 Smorodintsev Research Institute of Influenza, St. Petersburg, Russia. Electronic address: biomalinka@gmail.com. 2 Smorodintsev Research Institute of Influenza, St. Petersburg, Russia. 3 Smorodintsev Research Institute of Influenza, St. Petersburg, Russia; Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia. 4 Smorodintsev Research Institute of Influenza, St. Petersburg, Russia; Peter the Great St. Petersburg Polytechnic University, St. Petersburg, Russia.
[h=3]Abstract[/h] 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.
Copyright ? 2019. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Antibody array; Diagnosis of viral diseases; Monoclonal antibodies; Multiplex immunoassay; Multiplex molecular diagnostics; Severe acute respiratory infections
PMID: 31783022 DOI: 10.1016/j.jim.2019.112712