Rapid detection of herpes simplex virus in culture by in situ hybridization

Abstract

Detection of herpes simplex virus (HSV) by a newly developed HSV-DNA in situ hybridization (ISH) technique (Diagnostic Hybrids, Inc., Athens, OH, USA) was compared with a reference standard that combines observation for cytopathic effect in shell vial cultures with subsequent identification of virus by staining with fluorescein-labeled HSV-specific monoclonal antibody. The new technique utilizes a probe consisting of an alkaline phosphatase direct labeled, cloned, single stranded DNA fragment that is common to HSV-1 and HSV-2. Both methods include enhancement of infection by centrifugation. In concurrent testing of 98 freshly collected specimens, HSV was detected in 17 by culture and 16 by ISH. In testing of 57 frozen positive specimens, HSV was detected in 54 by culture and 53 by ISH. Of the total isolates, 22 were HSV-1 and 49 were HSV-2. HSV was detected after 24 hrs of incubation by the DNA probe technique. Using the shell vial method as a reference standard, the new HSV-DNA ISH method had a sensitivity of 97.2%, specificity of 100%, positive predictive value of 100%, and a negative predictive value of 97.7%. HSV-DNA ISH appears to be a practical, sensitive, and specific technique for detection of HSV.