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Journal of Clinical Microbiology, August 2005, p. 3779-3787, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.3779-3787.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

DNA Probe Array for the Simultaneous Identification of Herpesviruses, Enteroviruses, and Flaviviruses

Advanced Technology Unit, bioMérieux, 69280 Marcy l'Étoile, France,¹ CRSSA Emile Pardé, 24 avenue des Maquis du Grésivaudan, 38702 Grenoble, France²

Received 19 November 2004/ Returned for modification 1 February 2005/ Accepted 19 April 2005

Viral infections of the central nervous system (CNS) are caused by a variety of viruses, namely, herpesviruses, enteroviruses, and flaviviruses. The similar clinical signs provoked by these viruses make the diagnosis difficult. We report on the simultaneous detection of these major CNS pathogens using amplification by PCR and detection of amplified products using DNA microarray technology. Consensus primers were used for the amplification of all members of each genus. Sequences specific for the identification of each virus species were selected from the sequence alignments of each target gene and were synthesized on a high-density microarray. The amplified products were pooled, labeled, and cleaved, followed by hybridization on a single array. This method was successfully used to identify herpesviruses, namely, herpes simplex virus type 1 (HSV-1), HSV-2, and cytomegalovirus; all serotypes of human enteroviruses; and five flaviviruses (West Nile virus, dengue viruses, and Langat virus). This approach, which used highly conserved consensus primers for amplification and specific sequences for identification, would be extremely useful for the detection of variants and would probably help solve some unexplained cases of encephalitis. The analytical sensitivity of the method was shown to be 500 genome equivalents ml^–1 for HSV-1, 0.3 50% tissue culture infectious doses (TCID₅₀s) ml^–1 for the enterovirus coxsackievirus A9, and 200 TCID₅₀s ml^–1 for West Nile virus. The clinical sensitivity of this method must now be evaluated.

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