title: [Novel multiplex real-time PCR assay for detection and differentiation of herpes simplex virus type 1 and 2 DNA].
abstract: BACKGROUND
  Herpes simplex viruses type 1 and 2 are the cause of world spread multiple infections with different course and severity. The aim of this work was to design and to optimize multiplex real-time PCR assay for simultaneous detection of HSV-1 and HSV-2. The second aim of the project was to check if the designed method is laboratory useful analyzing different clinical specimens.
  
  
  METHODS
  In this experiment primers and probes were designed to specific viral sequences: for HSV-1 to the gene of viral DNA polymerase; for HSV-2 to the UL5 sequence. For performing qPCR assay TaqMan chemistry was used. Reference strains HSV-I McIntyre and HSV-2 MS were used as a positive control. To test laboratory utility of the designed method 58 different clinical specimens were analyzed.
  
  
  RESULTS
  Developed multiplex real-time PCR gave positive result only in the samples containing genetic material of HSV-1/2. Of the 58 clinical samples tested, 27 proved to be positive for HSV-1 and 17 for HSV-2. The 7 samples showed the presence of both types of DNA herpes simplex virus, and 7 others were found for both HSV-1 and HSV-2 negative.
  
  
  CONCLUSIONS
  Obtained results show that the designed method is highly specific and can possibly be used to simultaneously detect and differentiate HHV-1/2. Both high specificity and very short time of analysis have great importance in diagnosing immunocompromised patients, which ought to be diagnosed quickly and effectively in order to provide appropriate treatment.
authors.forename: Paulina
authors.initials: P
authors.lastname: Machura
authors.forename: Emilia
authors.initials: E
authors.lastname: Górka
authors.forename: Beata
authors.initials: B
authors.lastname: Młynarczyk-Bonikowska
authors.forename: Anna
authors.initials: A
authors.lastname: Majewska
authors.forename: Magdalena
authors.initials: M
authors.lastname: Malejczyk
authors.forename: Grażyna
authors.initials: G
authors.lastname: Młynarczyk
authors.forename: Tomasz
authors.initials: T
authors.lastname: Dzieciątkowski
mesh_terms: D002555:Cerebrospinal Fluid / Q000821:virology; D004279:DNA, Viral / Q000302:isolation & purification*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000302:isolation & purification*; D018258:Herpesvirus 2, Human / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D044967:Serum / Q000821:virology; D012867:Skin / Q000821:virology; D014556:Urine / Q000821:virology
chemical_list: D004279:DNA, Viral; D004259:DNA-Directed DNA Polymerase