Eye Fluid Viral PCR Quant Panel

The detection and quantitation of Cytomegalovirus (CMV), Herpes Simplex Virus (HSV), and Varicella Zoster Virus (VZV) in eye fluid using real-time PCR.

The detection and quantitation of CMV are performed using a modified FDA-approved Roche cobas® CMV Quantitative Assay, which allows for rapid identification through real-time PCR amplification. The lower limit of detection (LoD) and the lower limit of quantitation (LLoQ) are both 34.5 IU/mL. The LoD is defined as the lowest viral concentration that produces a positive result in 95 percent of replicates. The assay targets the UL54 gene using specific primers.

The detection and quantitation of HSV are performed using real-time PCR amplification. The assay identifies amplicons of the gB gene for both HSV-1 and HSV-2. Two distinct probes are used in the reaction, each specific to either HSV-1 or HSV-2, and are labeled with different reporter dyes. Quantitation of HSV DNA (copies/mL) is based on a standard curve generated from serial dilutions of a known concentration of DNA containing both HSV-1 and HSV-2 amplicons. The LoD and LLoQ are both 250 copies/mL (2.40 log copies/mL).

The detection and quantitation of VZV are also performed using real-time PCR amplification. The primers amplify a fragment of the ORF62 region. Quantitation is achieved using a standard curve prepared in each run from serial dilutions of a known amount of DNA containing VZV amplicons. The LoD and LLoQ are both 75 copies/mL (1.88 log copies/mL).

These tests were developed, and their performance characteristics determined, by the University of Washington Department of Laboratory Medicine and Pathology. They have not been cleared or approved by the U.S. Food and Drug Administration.