Rapid protocol of dot-immunnoassay for orthopoxviruses detection.

Rapid protocol of dot-immunnoassay for orthopoxviruses detection.
J Virol Methods. 2020 Mar 21;:113859
Authors: Poltavchenko AG, Ersh AV, Filatov PV, Yakubitskiy SN
Abstract
The aim of the work was to create a sensitive and fast immunochemical test for the detection of orthopoxviruses (OPXV) in the "point of care" format. This work presents the results of the comparative evaluation of a single-stage (rapid version) and two-stage protocol of dot-immunoassay based on plane protein array for detection of vaccinia virus (VACV), cowpoxvirus (CPXV) and ectromelia virus (ECTV) in viral culture materials with different degrees of purification. It has been established that rabbit polyclonal VACV-antibodies can be used in a one-stage dot-analysis, both as a capture agent immobilized on a substrate and as a detection reagent bound with colloidal gold particles. It is shown that the sensitivity of detection of OPXV is inversely related to the degree of purification of viruses. The one-stage variant of the dot-immunoassay allows reducing the analysis time to 40 minutes and increasing the detection sensitivity of all the studied orthopoxviruses in crude viral samples to a range of 104-103 PFU/ml. The increase in sensitivity in the rapid version of the analysis, presumably, occurs due to binding of capture antibodies to subviral structures that form large aggregates of gold particles. Ultrasonic treatment of culture virus reduces the detection se...

Source: Journal of Virological Methods - March 20, 2020 Category: Virology Authors: Poltavchenko AG, Ersh AV, Filatov PV, Yakubitskiy SN Tags: J Virol Methods Source Type: research