Comparison of simplexa HSV 1 & 2 PCR with culture, immunofluorescence and laboratory-developed taqman PCR for detection of herpes simplex virus in specimens

Abstract

The Simplexa HSV 1 & 2 direct PCR assay was compared with conventional cell culture, cytospin-enhanced direct fluorescent antibody (DFA), and a laboratory-developed real-time TaqMan PCR (LDT HSV PCR) using extracted nucleic acid for the detection of herpes simplex virus (HSV) in dermal, genital, mouth, ocular, and other swab samples.One hundred seventy-one swabs were tested prospectively, and 58 were positive for HSV (34 HSV-1 and 24 HSV-2).Cytospin-DFA detected 50 (86.2%), conventional cell culture 51 (87.9%), Simplexa direct 55 (94.8%), and LDT HSV PCR 57 (98.3%) of 58 true positives.Simplexa direct detected more positives than DFA and culture, but the differences were not significant (P-0.0736 and P-0.3711, respectively, by the McNemar test).Samples that were positive by all methods (n-48) were strong positives (LDT cycle threshold [CT] value, 14.4 to 26.1).One strongly positive sample was falsely negative by LDT HSV PCR due to a failure of TaqMan probe binding.Three samples falsely negative by Simplexa direct had high CT values by LDT HSV PCR (LDT CT, 35.8 to 38.2).Omission of the DNA extraction step by Simplexa direct led to a drop in sensitivity compared to the sensitivity of LDT HSV PCR using extracted samples (94.8% versus 98.3%, respectively), but the difference was not significant (P-0.6171).Simplexa HSV 1 & 2 direct PCR was the most expensive but required the least training of the assays used, had the lowest hands-on time and fastest assay time (75 min, versus 3 h by LDTHSVPCR), and provided the HSV type © 2013, American Society for Microbiology.All Rights Reserved.