ABSTRACT
PCR following two methods of DNA extraction was used to confirm the growth of camel pox virus (CPV) and vaccinia virus in cell culture and chorioallantoic membrane. Results were compared with the commonly used neutralization test. The first method of DNA extraction was accomplished by using viral DNA in tissue culture supernatant and Chorioallantoic membrane, which was released by initial heating for 15 min at 99 degrees C followed by ordinary PCR. In the second method DNA was extracted by using DNA Isolation Kit from tissue culture supernatant and used as a template. Rapid identification and differentiation of CPV and Vaccinia virus were achieved by PCR and this assay proved to be fast and feasible, and can be an alternative to orthodox serological methods.
Subject(s)
Orthopoxvirus/isolation & purification , Polymerase Chain Reaction/methods , Vaccinia virus/isolation & purification , Animals , Chlorocebus aethiops , Cytopathogenic Effect, Viral , Neutralization Tests , Vero CellsABSTRACT
Camel pox viruses isolated in Sudan and VD45 (African camel pox strain) and Vaccinia virus (Elstree strain) were used for inoculation of chorioallantoic membrane (CAM) of embryonated eggs (EE) and cell culture (CC). In EE Lesions were seen as pocks ranging in size from 1 to 1.5 mm in diameter, and they increase in size with serial passage and taking opaque- white and opaque- yellow colors. When propagated in Vero cells, these viruses gave clear CPE, characterized by rounding of cells, plaque formation, syncytia and detachment of cells from the glass.