EVALUATION OF H&V-MIXTM FRESH CELL FOR THE RAPID TURN-AROUND-TIME IN THE DETECTION OF VZV, HSV, AND CMV

Session ID:	S21
Author Name:	Cherry-Ann Da Costa, Carol Hamilton, Sara T. Beatrice, William R. Oleszko, and Maria Paz Carlos New York City Department of Health and Mental Hygiene, Public Health Laboratory, New York, New York
Country:	US
Conference Session:	Session I

Background: Isolation of Herpes simplex virus (HSV), Cytomegalovirus (CMV) and Varicella-Zoster virus (VZV) has been performed traditionally by conventional cell culture techniques. These techniques tend to vary in their incubation periods before cytopathic effect (CPE) can be visible from 18 hours in the case of HSV to as long as two weeks in the case of CMV and VZV. Centrifuged enhanced culture in shell vial allows a shorter incubation period and eliminates the necessity for the expertise required to read culture tubes. Usually different cell lines are needed for these specific viruses but it is now possible to inoculate into a single mixed cell system to achieve similar, if not better results.

Objective: The primary objective of this study is to determine the benefit of using

H&V-Mix™ Fresh cells, Diagnostic Hybrids Inc. (DHI) to aid in the rapid diagnosis of HSV, CMV and VZV in a single test system. A comparison therefore is made using

A-549 cell lines for the isolation of HSV and VZV and MRC-5 for the isolation of CMV.

Method: A commercial panel of isolates with known results (DHI) containing a mixture of CMV, HSV, VZV was used in this study. H&V Mix™ cells were inoculated in triplicate. The first batch was incubated for an interval of 24 hours then fixed and stained with HSV antibody. The second and third batches were incubated for 72 hours then fixed and stained with VZV and CMV (early) antibody respectively. In addition A-549 shell vials were inoculated in duplicate and stained with HSV antibody after 24 hours and VZV antibody after 72 hours. A third batch was also inoculated into MRC-5 shell vials and stained CMV (early) antibody after 72 hours.

Results: H&V Mix™ was not only able to detect all positives when compared to the single cells but the intensity of fluorescence in the mixed cell system was stronger when compare to the single cell systems.

Conclusion: The overall effect of inoculating specimens of unknown etiology (HSV, VZV, CMV) with similar clinical manifestations into a single cell system(H&V Mix™) capable of rapid laboratory diagnosis is not only cost effective but also can provide faster results as a differential diagnosis.