These specimens are routinely set up for herpes simplex virus only. Other viruses such as enteroviruses will be looked for only if specifically requested. Refer to Appendix XV (Virus Isolation and Identification) to ensure that the appropriate media is inoculated.
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II. Collection and Transport
Mouth or throat washings are collected using sterile saline and placed in a clean, sterile container and transported to the laboratory as soon as possible. If a delay in transport or processing is anticipated, keep the specimen at 4oC.
III. Procedure
A. Processing of Specimen:
a) Transfer 2mL of specimen to a sterile freezer vial.
b) Add 0.2 mL (4 drops) of gentamicin and 0.1 mL (2 drops) of fungizone to a final concentration of 100 mg/mL and 10 mg/mL respectively.
c) Let stand at room temperature for 10 minutes.
B. Direct Examination:
Not done.
C. Isolation and Identification:
| Method | Cell Linesa | Incubation at 36oC | Stainb used |
| Shell Vial | MRC-5 MRC-5 | 1 day 1 day | HSV1 HSV2 |
aMRC-5 = Human diploid fibroblast cells
b HSV1= Monoclonal antibody DFA stain for Herpes simplex 1
b HSV2= Monoclonal antibody DFA stain for Herpes simplex 2
Refer to Appendix II for Shell Vial staining and interpretation.
IV. Reporting Results
Shell Vial: Negative Report: “Negative for Herpes Simplex virus”
Positive Report*: “POSITIVE for Herpes Simplex virus type____.”
V. Reference
1. Isenberg, HO. 1992. Clinical Microbiology Procedure Handbook, Vol. 2. ASM.
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