A New Isolated Local Varicella Virus: Isolation, Identification, Comparative Growth Characteristics and Immunological Evaluation in an Animal Model

Abstract

A panel of 4 different cell lines was optimized for isolation, identification, and authentication of a varicella zoster virus from a swab sample of an 8-year-old boy suspected to varicella zoster infection. The system enabled highly efficient and rapid isolation of viruses in 33°C by serial sub culturing to more than 25 passages. The technique relies on isolation of viral genes by increasing the number of particles that are statistically represented in cell culture and verified by cell culture infectious dose 50% assay, fluorescence amidites- real time- polymerase chain reaction, and the varicella-zoster virus immediate-early protein 62 antibody in immunofluorescence test, using vaccinal Oka as attenuated varicella zoster virus golden standard. The viral genes (ORF38, ORF54) confirmed the new isolate as varicella zoster virus and revealed the amino acid sequence of viral-encoded proteins after 27 passages, identical with positive control virus, in restriction fragment length polymorphism polymerase chain reaction test. Utilization of successive serial passages at temperatures lower than the normal body temperature would reduce the virus virulence and directly cause virus attenuation