Colorzyme EA, A simple and rapid test for identification of Herpesviridae and enteroviruses in infected cell culture

ABSTRACT

In this study, the Immuno-concepts EA, a new indirect enzyme antibody technique was used. In this technique, diluted patient sera was added to a fixed spoted slide containing 10% viral infected cells which was incubated 30 minutes at room temperature [R1]. After washing, antihuman conjugate peroxidase was added for further 30 minutes and washed then enzyme substrate was added for the same period, washed that produces a visible color reaction instead of fluorescent labeled one and only the slides were dipped 10 minutes in counter stain and was examined by ordinary microscope, In positive samples, 10% of the infected cells gave a dark blue-purple staining of the entire cell or cytoplasm according to the virus present while the negative cells showed faint red color. This test was commercially produced by lmmuno-Concepts, U.S.A. to detect Ig[G] antibodies in patient sera. When we used colorzyme test for virus identification, the results were very good. When patient samples were inoculated into cell culture grown in Lab. Tec tissue culture [TC] chamber slides or TC flasks and when CPE was detected, the slide was tested by colorzyme test. Results of Herpes simplex and cytomegalovirus [CMV] virus identification was identical with immuno fluorescent [IF] test and the same observation was detected in the samples containing polioviruses and the results of neutralization test was the same like colorzyme test. All incubations in colorzyme test was in room temperature and only an ordinary microscope was used instead of fluorescent microscope used in [IF] test or plate washers and readers in Elisa beside incubators. Our results indicate that colorzyme immuno concept was a simple, rapid and sensitive test for viral diagnoses and can be used in any private laboratory