title: Detection of a transforming fragment of herpes simplex virus type 2 in clinical specimens by PCR. The Canadian Women's HIV Study Group.
abstract: A PCR assay for the sensitive detection of a transforming fragment of herpes simplex virus type 2 (HSV-2) was developed. Oligonucleotide primers were selected in Xho-2, a transforming region of the BglII N fragment of HSV-2. The assay reached a sensitivity endpoint of 10 copies of the Xho-2 subfragment and did not show cross-reactivity with other herpesviruses, including HSV-1. All 42 HSV-2 isolates scored positive in the assay. The Xho-2 PCR assay was evaluated with 216 clinical specimens and the results were compared with those of cell culture. The best protocol for processing specimens contained in viral transport medium included a centrifugation step followed by cell lysis. Of the 107 specimens positive for HSV-2 by culture, 105 were PCR positive (sensitivity, 98.1%). For one of the two falsely negative samples, beta-globin as well as sequences from the HSV-2 DNA polymerase gene could not be amplified. The other sample scored positive in both of these reactions but was indeterminate in duplicate tests by Xho-2 PCR. Two of 109 HSV-2 culture-negative specimens tested positive in the PCR assay (specificity, 98.2%). The latter two samples tested positive in a PCR test for the HSV-2 DNA polymerase gene. This novel tool was shown to be sensitive and specific for HSV-2 sequences and should allow for the investigation of the role of HSV-2 in genital cancers.
authors.affiliation: Département de Microbiologie-Immunologie, Université de Montréal, Québec, Canada.
authors.forename: G H
authors.initials: GH
authors.lastname: Guibinga
authors.forename: F
authors.initials: F
authors.lastname: Coutlée
authors.forename: A
authors.initials: A
authors.lastname: Kessous
authors.forename: C
authors.initials: C
authors.lastname: Hankins
authors.forename: N
authors.initials: N
authors.lastname: Lapointe
authors.forename: G
authors.initials: G
authors.lastname: Richer
authors.forename: J
authors.initials: J
authors.lastname: Tousignant
mesh_terms: D000328:Adult; D001483:Base Sequence; D015342:DNA Probes / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005069:Evaluation Studies as Topic; D005260:Female; D006558:Herpes Genitalis / Q000175:diagnosis / Q000821:virology; D018258:Herpesvirus 2, Human / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D014170:Transformation, Genetic
chemical_list: D015342:DNA Probes; D004279:DNA, Viral