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Suitability and clinical application of a multiplex nested PCR assay for the diagnosis of herpes simplex virus infections
Author(s) -
Cassinotti P.,
Mietz H.,
Siegl G.
Publication year - 1996
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/(sici)1096-9071(199609)50:1<75::aid-jmv13>3.0.co;2-x
Subject(s) - herpes simplex virus , virology , polymerase chain reaction , nested polymerase chain reaction , multiplex polymerase chain reaction , virus , biology , dna extraction , herpesviridae , alphaherpesvirinae , multiplex , hsl and hsv , medicine , microbiology and biotechnology , viral disease , gene , bioinformatics , biochemistry
A novel multiplex nested polymerase chain reaction (PCR) assay was designed and evaluated for routine diagnosis of herpes simplex virus (HSV) infections in patients with either putative HSV infection of the central nervous system or suspected HSV keratitis. Single‐tube amplification of HSV type 1 (HSV‐1) or type 2 (HSV‐2) DNA extracted from cerebrospinal fluid (CSF) or from keratectomy specimens was followed by differentiation of the virus type‐specific PCR products either by agarose gel analysis or by DNA enzyme immunoassay. Among 417 CSF specimens obtained from 395 consecutive patients with clinically suspected HSV infection, 11 (2.6%) were positive for HSV‐1 DNA and four (1.0%) probes were positive for HSV‐2 DNA. None of the specimens was positive for both HSV‐1 and HSV‐2 DNA. The genome of HSV‐2 was detected in a CSF sample obtained from a woman with meningoencephalitis and genital herpes. The presence of PCR inhibitors was detected in six of 111 (5.4%) reconstructed CSF samples. Inhibition could be removed following extraction with a commercial kit. HSV‐1 DNA, but no HSV‐2 DNA, was detected in corneal buttons obtained from patients with suspected herpetic keratitis. No contamination has been recorded during the 2‐year routine use of this test, which has met the specific requirements of a diagnostic laboratory. © 1996 Wiley‐Liss, Inc.