Retrospective evaluation of the isolation and identification of herpes simplex virus with Cultureset and human fibroblasts

Abstract

A total of 442 specimens from various anatomic sites was cultured for herpes simplex virus during the past 2 years. Most specimens were obtained from the respiratory tracts and cutaneous lesions of immunocompromised hosts (232 specimens) or the female genital tract (138 specimens). Two tubes containing human newborn foreskin fibroblasts and two Ortho Cultureset tubes containing Vero cells were inoculated with each specimen. The 384 inoculated specimens were stained with Cultureset peroxidase-antiperoxidase reagents within 48 h and again at 3, 4, or 5 days if initially negative. Fibroblasts were inspected for cytopathic effect for 7 days. Of these 384 specimens, Cultureset detected 57 of 62 positive specimens within 48 h; fibroblasts detected 58 positive specimens by 7 days. The calculated sensitivity and specificity for Cultureset at 48 h were 91.9 and 100%, respectively. However, when all results were considered, including those that became positive in Cultureset after 48 h, the calculated sensitivity and specificity for Cultureset were 98.8 and 100%, respectively. We conclude that Cultureset is a reliable method for detection of herpes simplex virus when two tubes are inoculated and stained as described.

Viral isolation versus immune staining of infected cell cultures for the laboratory diagnosis of herpes simplex virus infections

“…The Difco Cellmatics kit was found to be effective for detecting HSV from a variety of clinical specimens, and results from this study are as good as or better than results published for studies with similar kits (4,6,9,16,17,19). However, we found that conventional culturing detected more HSV isolates than did culturing followed by staining (146 versus 138).…”

“…However, it is important to note that conditions were not identical, because different specimens were used with each cell type. Comparative studies will therefore be necessary to determine whether Vero cells are acceptable as hosts for HSV detection, especially because several commercial kits come with this cell line and because differences in sensitivity have been reported (4,6,9,16,17,19). We were able to rapidly detect and report 66% (96 of 146) of the HSV-positive specimens within 24 h by a standard viral culture method.…”

“…Immunoperoxidase and peroxidase-antiperoxidase staining can be an effective means for rapidly identifying HSV antigens either in infected cell cultures or directly in clinical specimens (1,2,11,12,14,15,18). Recently, several commercial HSV detection kits involving immunoperoxidase staining have been evaluated with various degrees of success (4,6,9,16,17,19). In the present report, we document our experience for detecting HSV in 458 clinical specimens by either conventional cell culture isolation or immunoperoxidase staining after viral amplification in culture.…”

Impact of cell culture sensitivity and virus concentration on rapid detection of herpes simplex virus by cytopathic effects and immunoperoxidase staining

“…16, 10, 7, and 0 were positive in ML, RK, MRC-5, and Vero cells, respectively. On day 2 p.i., CPE was revealed in ML cells for 21 specimens, as compared with 20 for RK, 16 for MRC-5, and 3 for Vero cells. By day 5, all 22 specimens were reisolated in ML cells, 21 in RK cells, 17 in MRC-5 cells, and only 14 in Vero cells.…”

Physical and chemical methods for enhancing rapid detection of viruses and other agents.

“…HSV can also be rapidly detected in noncentrifuged cultures by immune staining (65,116,128,137,152,180,187,192,202,203,219,239,243,257,344). Most studies that use commercial immunoperoxidase staining kits and noncentrifuged cell cultures have found immunoperoxidase staining at <48 h to be less sensitive than conventional cultures for detecting HSV (65,116,128,239,243,257).…”