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T. Deai, M. Fukuda, T. Hibino, S. Higaki, Y. Shimomura, K. Hayashi; Quantitation of HSV-DNA in Clinical Samples Obtained From Various Herpetic Eye Diseases . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4280.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To understand the pathogenesis of herpetic eye diseases, we quantitated herpes simplex virus (HSV) genomes in clinical samples (tear film, aqueous humor and corneal tissue) obtained from various herpetic eye diseases. Methods: We sampled tear film from various herpetic keratitis; epithelial keratitis (HEK)(38eyes/38patients), active stromal keratitis (HSK)(22/22), silent HSK (8/8), endotheliitis (3/3), persistent epithelial defect (PED)(9/9), aqueous humor from uveitis (6/6) and host corneas obtained from the patients who had past history of herpetic keratitis (6/6). ABI PRISMTM7700 Sequence Detector (PE Applied Biosystems) was used for a real-time PCR assay. Results: This assay had a wide linear range from 1 to 107 copies. The values of coefficient of variation intra- and inter-assay were 3.0% and 10.1%, respectively. HSV-DNA were detected in 81.5% of HEK patients (3.9×106 copies/sample in an average), 59.1% of active HSK patients (8.9×105), none in silent HSK and endotheliitis, 88.9% of PED patients (9.2×104), 16.7% of uveitis (3.82×105 copies/ml) and 83.3% of corneas (1.9×104 copies/mg). The amount of HSV-DNA in HEK patients was significantly higher than that in active HSK (P<0.05). After clinical amelioration by acyclovir, the copy numbers became below the sensitivity level of this method. Conclusions: HSV-DNA was detected in active HSK, PED in tear samples and the host corneas having past history of HSV keratitis as well as HEK. A real-time PCR is a useful method for quantitatiation of HSV genome in herpetic eye diseases.
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