Isolation of the Herpes Simplex Virus by Shell Vial Culture

Choi, Tae Y; Lee, Kyu T; Park, Hae I; Kang, Jung O

Abstract

BACKGROUND
Cell culture is the golden standard method for Herpes simplex virus (HSV) isolation. However, some specimens require many days to develop any cytopathic effect (CPE). We developeda rapid sensitive culture technique for HSV isolations. METHODS: This study included a total of 133 patients with suspected HSV infection. Specimens were centrifuged onto a Vero cell monolayer in a shell vial. The CPE was observed daily during the5-day incubation by inverted-phase microscope. The direct immunofluorescence (DIF) stain with aHSV specific antibody was performed 2 days after sample inoculation. The negative samples in theDIF stain were reinoculated in the new shell vials after extraction of the monolayer. Polymerase chainreaction for HSV detection was performed using the original samples. RESULTS: The CPE was observed 30 (64%), 39 (83%), 43 (92%), 44 (94%), and 46 (98%) cases at1, 2, 3, 4, and 5 days incubation, respectively. The DIF stain detected 46 cases (98%) at 2 days incubation. The CPE was observed in another 7 cases at 1-day incubation after the reinoculation of negative samples. The PCR detected 47 (100%) of 133 cases. CONCLUSIONS: The reinoculation of negative sample in a shell vial culture is a rapid sensitive methodfor HSV isolation.