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

Abstract

The Difco Laboratories Celimatics kit, an immune staining method for the detection of herpes simplex virus after amplification in culture, was found to be 94.6% (138 of 146) sensitive and 100% (302 of 302) specific. However, conventional culturing detected 10 (6.4%) additional viruses that were not herpes simplex. For a full-service diagnostic laboratory, viral isolation is important.

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

“…Time and materials are also saved since the cultures are terminated at 48 h instead of the 5 to 7 days recommended for conventional HSV cultures (13). One disadvantage is the lack of detection of other viruses which may be in the specimens (10).…”

“…A number of studies have compared these commercial kits with conventional isolation in cell culture (4,8,10,11,16,17,18,20,21). In all except two reports (11,20), different cell cultures were used for the two methods ( Table 5).…”

“…This discrepancy might be explained by lower sensitivity of the strain of human cells used, differences in laboratory methods, and possibly less experience in conventional viral diagnostic techniques. Kits utilizing cell systems other than Vero cells have compared more favorably with conventional virus isolation (10,20). More recently, centrifugation of specimens onto cell monolayers in shell vials has been shown to significantly enhance the rapidity and sensitivity of HSV detection, particularly when used in conjunction with IP staining (7,20).…”

“…Most studies comparing commercial IP staining kits with conventional cell culture techniques have found IP staining at 48 h to be less sensitive than cultures observed for CPE for 7 to 10 days (4,8,10,17,18,21). It has been postulated but not established, that low titers of virus in the specimens account for some of the discrepancies between the two techniques.…”

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). The sensitivity for detecting HSV in noncentrifuged cultures by immune staining has been variable and has ranged from 48 to 100% (65,187,192,202,203,243).…”