title: Comparison of two methods of PCR followed by enzymatic restriction digestion for detection and typing of herpes simplex viruses isolated from patients with mucocutaneous or cutaneous lesions.
abstract: A multitude of different polymerase chain reactions (PCRs) have been described for detection and typing of Herpes simplex virus (HSV). This paper compares two PCRs coupled to enzymatic restriction (PCR/RFLP) to detect and type HSV. A primers set was designed to amplify a HSV DNA fragment from UL30 and UL 15 genes. Typing was done by restriction of the UL30 and UL15 amplicons with Ava II and Hpa II enzymes, respectively. This strategy was tested with two reference strains (HSV-1 McIntyre, and HSV-2 G), and 47 clinical HSV isolates. Both PCRs produced the expected amplicons (a 492 bp UL30, and 305 bp UL15). The restriction of both amplicons clearly differentiated HSV- from HSV-2, and produced equal results. Thirty one (66%) of the isolates were identified as HSV-1, and the other 16 (34%), as HSV-2. Most of the HSV-1 isolates (27/31) were from orofacial and thoracic lesions; and also, one half of the HSV-2 isolates (8/16) were from the same anatomical regions. Our results showed that either of the two PCR/RFLP could be used to detect and type HSV. Furthermore, our results of the anatomical site of HSV-1 and HSV-2 infections are consistent with previous reports which have shown changes in the classical anatomical localization of herpesvirus infections.
authors.affiliation: Laboratorio de Virología, ENCB-IPN, Casco de Santo Tomás, DF México.
authors.forename: Emma
authors.initials: E
authors.lastname: Herrera-Martínez
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Ondarza-Aguilera
authors.forename: Sergio
authors.initials: S
authors.lastname: Estrada-Parra
authors.forename: Guillermo
authors.initials: G
authors.lastname: Pérez
authors.forename: Blanca L
authors.initials: BL
authors.lastname: Barrón
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D004279:DNA, Viral / Q000302:isolation & purification; D006558:Herpes Genitalis / Q000821:virology; D006561:Herpes Simplex / Q000821:virology*; D018259:Herpesvirus 1, Human / Q000145:classification / Q000302:isolation & purification*; D018258:Herpesvirus 2, Human / Q000145:classification / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D014709:Vero Cells; D014773:Virology / Q000379:methods*
chemical_list: D004279:DNA, Viral