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Journal of Clinical Microbiology, November 1999, p. 3518-3523, Vol. 37, No. 11
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Amplification of Reiterated Sequences of Herpes Simplex Virus Type 1 (HSV-1) Genome To Discriminate between Clinical HSV-1 Isolates

J. Maertzdorf,1 L. Remeijer,2 A. Van Der Lelij,3 J. Buitenwerf,4 H. G. M. Niesters,1 A. D. M. E. Osterhaus,1 and G. M. G. M. Verjans1,2,*

Institute of Virology, Erasmus University Rotterdam, 3000 DR Rotterdam,1 The Rotterdam Eye Hospital, 3000 LM Rotterdam,2 Department of Ophthalmology, University Hospital Utrecht, 3508 GA Utrecht,3 and Zuider Ziekenhuis, 3011 EN Rotterdam,4 The Netherlands

Received 27 April 1999/Returned for modification 4 June 1999/Accepted 23 July 1999

Herpes simplex virus type 1 (HSV-1)-related disease ranges from a localized, self-limiting illness to fatal disease in immunocompromised individuals. The corneal disease herpetic keratitis may develop after reactivation of a latent virus or reinfection with an exogenous herpesvirus. Molecular analysis of the virus involved may allow distinction between these two options. The HSV-1 genome contains several hypervariable regions that vary in numbers of reiterating regions (reiterations I to VIII [ReI to ReVIII]) between individual strains. Twenty-four HSV-1 clones, derived by subcloning of HSV-1 (strain F) twice in limiting dilutions, were tested in a PCR-based assay to analyze the stabilities of ReI, ReIII, ReIV, and ReVII. ReI and ReIII proved to vary in size upon subcloning, whereas ReIV and ReVII were stable. Subsequently, 37 unrelated isolates and 10 sequential isolates from five patients, all with HSV-1-induced keratitis, were genotyped for ReIV and ReVII. Of the 37 unrelated samples, 34 (92%) could be discriminated, while the genotypes of the viruses in sequential samples were identical for each individual. Conclusively, the data show that the approach presented allows the rapid and accurate discrimination of HSV-1 strains in studies that address the transmission and pathogenesis of HSV-1 infections.

* Corresponding author. Mailing address: Department of Virology, Erasmus University Rotterdam, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands. Phone: (31) 10-4088066. Fax: (31) 10-4089485. E-mail: verjans{at}viro.fgg.eur.nl.

Journal of Clinical Microbiology, November 1999, p. 3518-3523, Vol. 37, No. 11
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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