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Detection of the six commonest human herpesviruses in clinical specimens by a single PCR (Herpes Consensus methodology)

Abstract number: 902_p845

Levidiotou S.

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Objectives:

A wide spectrum of clinical disease is associated with the human herpesviruses. The six most frequent aetiologic agents of herpetic infections are herpes simplex virus 1 and 2 (HSV1, HSV2), varicella-zoster virus (VZV), cytomegalovirus (CMV), Epstein–Barr virus (EBV) and human herpesvirus 6 (HHV-6). The aim of this study was the simultaneous detection and identification of the six major human herpesviruses in various clinical specimens by a single PCR.

Methods:

A total of 136 clinical samples obtained from 135 patients, including 78 specimens of cerebrospinal fluid (CSF) from patients with suspected encephalitis or meningitis, six swabs from patients with vesicular skin lesions, 41 aqueous humor specimens from patients with uveitis or iridocyclitis, nine conjunctival swabs from patients with conjunctivitis, and one vesicle aspirate and one bronchoalveolar lavage from the same patient with suspected varicella pneumonitis, were studied for the presence of herpesvirus DNA. These specimens were collected from March 2001 to November 2003. DNAs were amplified using a new type of primer system, ‘stair primer’, in a single tube, bringing simultaneous amplification of the six Herpesviruses (Herpes Consensus Hybridowell, Argene Biosoft, France). The amplified product was detected by hybridisation in a microtitre plate with six different biotinylated probes, specific for the six viruses. The method was successfully tested with positive and negative controls from the European Union Concerted Action for Quality Control of Nucleic Acid Amplification in Diagnostic Virology (Herpes-2001 and VZV-2003 Proficiency Panels). According to our results: (a) HSV1 DNA was detected in one CSF specimen from a patient with aseptic meningitis, in three aqueous humor specimens from patients with uveitis, in one swab from a patient with herpetic vesicular skin lesions and in three conjunctival swabs from patients with conjunctivitis, and (b) VZV DNA was detected in two aqueous humor specimens from patients with iridocyclitis, in two swabs from patients with vesicular skin lesions, and in the vesicle aspirate and bronchoalveolar lavage from the patient with varicella pneumonitis. The precise diagnosis of herpetic infection was available within 24–48 h, which allowed for an early initiation of adapted antiviral therapy.

Conclusion:

The detection of the six commonest human herpesviruses in clinical specimens by the Herpesvirus Consensus PCR methodology allowed rapid, sensitive and specific results.

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Session Details

Date: 01/08/2007
Time: 00:00-00:00
Session name: XXIst ISTH Congress
Subject:
Location: Oxford, UK
Presentation type:
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