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COMPARISON OF DIAGNOSTIC HYBRIDS NEW D3 HERPES SIMPLEX VIRUS DETECTION KIT WITH SYVA MICROTRAK FOR CULTURE CONFIRMATION Joan Barenfanger, Janet O’Brien, Tina Mueller, Memorial Medical Center, Springfield, IL.

Abstract Background: Isolation of Herpes simplex (HSV) from clinical specimens utilizing standard tube cell culture requires the formation of CPE which can take up to 7-days to develop. HSV does not have a diagnostic CPE and all potential positive cultures need to be confirmed. This is accomplished by staining cells expressing CPE with virus-specific monoclonal antibody reagents. This study evaluates a new 15-minute stain that has been developed by Diagnostic Hybrids, Inc. (Athens, OH [DHI]). Objective: The goal of this study was to compare the new DHI D3 HSV stain with the MicroTrak® HSV 1/HSV 2 stain (Trinity Biotech, Carlsbad, CA and Wicklow, Ireland). The stains were used for culture confirmation of either infected MRC-5 or A-549 cells (DHI). Design/Methods: A total of 107 specimens received in viral transport were included in the study. The specimens were inoculated into an A549 and MRC-5 tube culture. The cultures were incubated for up to 5days or until CPE was observed. The culture tubes showing CPE were processed according to each manufacturer’s package insert. The processed cell spots were then fixed, stained and read according to each manufacturer’s package insert. Results: There were no discrepant results between the two reagents. The new The D3 HSV reagent showed increased fluorescence in some cultures, but the intensity of both reagents was sufficient to identify all of the positives.

Methods A total of 107 specimens received in viral transport were included in the study 1. The specimens were inoculated into a MRC-5 and A549 tube culture. 2. The cultures were incubated at 35°C for up to 5-days or until CPE was observed. 3. Duplicate cell spots were made from the culture tubes showing CPE. 4. The processed cell spots were then fixed in acetone for 10-minutes. 5. The cell spots were stained according to each manufacturer’s package insert: DHI 15-minutes, Trinity 30-minutes. 6. The cells were interpreted for both the number of positive cells and the intensity of the fluorescence.

Results There were no discrepant results between the two reagents. The new The D3 HSV reagent showed increased fluorescence in some cultures, but the intensity of both reagents was sufficient to identify all of the positives. There were 35 HSV-1 and 28 HSV-2 isolated. The reagent stained both types with equivalent intensity.

Conclusion: The D3 HSV reagent produced comparable results when used for culture confirmation. It identifies both HSV-1 and HSV-2 equally well, and the 15-minute staining time saves time. DHI D3 HSV Stain

There were 35 HSV-1 and 28 HSV-2 isolated. The reagent stained both types with equivalent intensity. Conclusions: The D3 HSV reagent produced comparable results when used for culture confirmation. It identifies both HSV-1 and HSV-2 equally well, and the 15-minute staining time saves time.

Background Isolation of Herpes simplex (HSV) from clinical specimens utilizing standard tube cell culture requires the formation of CPE which can take up to 7-days to develop. HSV does not have a diagnostic CPE and all potential positive cultures need to be confirmed. This is accomplished by staining cells expressing CPE with virus-specific monoclonal antibody reagents. This study evaluates a new 15-minute stain that has been developed by Diagnostic Hybrids, Inc. (Athens, OH [DHI]).

Photographs provided by Diagnostic Hybrids, Inc.

Syva HSV Stain


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