Day 1 :
Research Centre of Biotechnology of the Russian Academy of Sciences, Russia
Alexandr Urusov has completed his PhD in A.N. Bach Institute of Biochemistry of the Russian Acad. Sci. He is the head of the group of bioanalytical systems in Research Center of Biotechnology of the Russian Acad. Sci. He has published 36 papers in reputed journals focused on different aspects of immunochemical interactions and features of immunodetection for compounds important for medicine and food safety.
Adenoviral infection is a very common viral disease. Its clinical manifestations include lesions of the respiratory system or the gastrointestinal tract. Main techniques used for the detection of adenovirus are ELISA and PCR. They are effective, but time-consuming, require highly skilled specialists and expensive equipment. These limitations are overcoming by immunochromatographic assay. The aim of the presented study was to create an test for rapid detection of adenovirus in stool samples.
Monoclonal antibodies A46 (Artron, Canada) were selected based on a comparison of 12 antibody preparations from different manufacturers. Gold nanoparticles (38 ± 3 nm) are chosen as a marker which provides high staining intensity and high capacity of immunoglobulins sorption. Conjugates of the nanoparticles with antibodies were characterized by dynamic light scattering and demonstrated the average diameter of 65 ± 6 nm. Immune reactivity of immobilized antibodies was confirmed.
The formed set of reactants was used to prepare multimembrane immunochromatograpic composites. The CNPC-15 membrane (MDI, India) was chosen as the optimal working membrane. It simultaneously gives a high specific coloration and acceptable speed of sample movement. The prepared test strips were checked for adenovirus control in stool samples. To eliminate non-specific binding, the addition of Tween-20 and BSA to the working was proposed; their optimal concentrations were chosen.
The required sensitivity of 30 ng/ml is confirmed. The short time of the assay (10 min) and simple point-of-care assessment of the obtained results provide competitive advantages of the developed tests for clinical diagnostics.