title: Detection of herpes simplex and varicella-zoster virus in clinical specimens by multiplex real-time PCR and melting curve analysis.
abstract: Herpes simplex viruses types 1 and 2 (HSV-1 and HSV-2), and varicella-zoster virus (VZV) are common agents resulting in various forms of clinical manifestation from skin vesicle to disseminated viral infection. The aim of the present study was to develop a real-time PCR and melting curve analysis which detect and differentiate HSV-1, HSV-2, and VZV, to compare with PCR-RFLP using clinical specimens, and to introduce the 4-year experience in the clinical laboratory. Three pairs of primers for HSV-1, HSV-2, and VZV were designed. Primers for human endogenous retrovirus-3 (HERV-3), an internal control, were adopted. A hundred selected specimens and many clinical specimens were tested for methods comparison and assay validation. Increased sensitivity and specificity were obtained from real-time PCR. In review of results of clinical specimens submitted to clinical laboratory, a total of 46 of 3,513 specimens were positive in cerebrospinal fluids, blood, skin vesicles, genital swabs, aqueous humor, and ear discharge. Thus, this method could be a rapid and accurate alternative to virus culture and other molecular tests for detection and typing of HSV-1, HSV-2, and VZV.
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul 110-744, Republic of Korea ; Department of Laboratory Medicine, Seoul National University Bundang Hospital, 173-82 Gumiro, Bundanggu, Seongnam, Gyeonggido 463-707, Republic of Korea.
authors.forename: Yun Ji
authors.identifier: 0000-0001-9163-5017
authors.initials: YJ
authors.lastname: Hong
authors.affiliation: Department of Laboratory Medicine, Seoul National University Bundang Hospital, 173-82 Gumiro, Bundanggu, Seongnam, Gyeonggido 463-707, Republic of Korea.
authors.forename: Mi Suk
authors.initials: MS
authors.lastname: Lim
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul 110-744, Republic of Korea ; Department of Laboratory Medicine, Seoul National University Bundang Hospital, 173-82 Gumiro, Bundanggu, Seongnam, Gyeonggido 463-707, Republic of Korea.
authors.forename: Sang Mee
authors.identifier: 0000-0003-3390-1932
authors.initials: SM
authors.lastname: Hwang
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul 110-744, Republic of Korea ; Department of Laboratory Medicine, Seoul National University Bundang Hospital, 173-82 Gumiro, Bundanggu, Seongnam, Gyeonggido 463-707, Republic of Korea.
authors.forename: Taek Soo
authors.identifier: 0000-0002-2093-1721
authors.initials: TS
authors.lastname: Kim
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul 110-744, Republic of Korea ; Department of Laboratory Medicine, Seoul National University Bundang Hospital, 173-82 Gumiro, Bundanggu, Seongnam, Gyeonggido 463-707, Republic of Korea.
authors.forename: Kyoung Un
authors.identifier: 0000-0002-2402-7633
authors.initials: KU
authors.lastname: Park
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul 110-744, Republic of Korea ; Department of Laboratory Medicine, Seoul National University Bundang Hospital, 173-82 Gumiro, Bundanggu, Seongnam, Gyeonggido 463-707, Republic of Korea.
authors.forename: Junghan
authors.initials: J
authors.lastname: Song
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul 110-744, Republic of Korea.
authors.forename: Eui Chong
authors.initials: EC
authors.lastname: Kim
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002460:Cell Line; D005260:Female; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000821:virology; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000302:isolation & purification; D018258:Herpesvirus 2, Human / Q000235:genetics* / Q000302:isolation & purification; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009691:Nucleic Acid Denaturation; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D012367:RNA, Viral