The ELVIS HSV Id test kit (an enzyme-linked virus-inducible system) (Diagnostic Hybrids, Inc.) uses genetically engineered BHK cells to produce a detectable enzyme, beta-galactosidase, upon infection with either herpes simplex virus (HSV) type 1 (HSV-1) or HSV-2. Twenty six ELVIS-positive clinical specimens were selected for study by PCR and with monoclonal antibodies because they were originally low-titer HSV-positive specimens by ELVIS but HSV antibody nonreactive upon follow-up staining of the ELVIS monolayer. Twenty-one of 26 specimens were frozen, thawed, and retested with ELVIS without removing the cellular debris from the specimen; 18 were ELVIS positive and 3 were ELVIS negative on retesting. A typing result was provided upon retesting for 14 of 18 ELVIS-positive specimens (11 were HSV-1 and 3 were HSV-2) with HSV-specific monoclonal antibodies; no antibody signal was observed for 4 of 18 ELVIS-positive specimens. Sixteen of 26 specimens were subjected to blinded PCR analysis with two different primer sets, including all those that were repeat tested with ELVIS without success and those that had insufficient quantity for repeat testing. All 16 specimens analyzed were PCR positive with primer set 1; 15 of 16 were also positive with primer set 2, with the HSV type identified for all specimens (7 were HSV-1 and 8 were HSV-2). These results indicate that the original ELVIS result with these low-titer specimens was correct and further confirm the sensitivity and specificity of ELVIS HSV Id as a rapid, cell culture-based kit for the detection of HSV.