title: [REAL TIME POLYMERASE CHAIN REACTION IN TULAREMIA LABORATORY DIAGNOSTICS].
abstract: OBJECTIVE
  Enhancement of tularemia laboratory diagnostics by F. tularensis DNA determination in blood sera of patients using real time polymerase chain reaction (RT-PCR).
  
  
  METHODS
  39 blood sera of patients obtained during transmissive epidemic outbreak of tularemia in Khanty-Mansiysk in 2013 were studied in agglutination reaction, passive hemagglutination, RT-PCR. Specific primers and fluorescent probes were used: ISFTu2F/R+ISFTu2P, Tu14GF/R+tul4-PR2.
  
  
  RESULTS
  Advantages of using RT-PCR for early diagnostics of tularemia, when specific antibodies are not detected using traditional immunologic methods, were established. Use of a combination of primers and ISFTu2F/R+ISFTu2P probe allowed to detect F. tularensis DNA in 100% of sera, whereas Tul4G F/R+tul4-PR2 combination--92% of sera. The data were obtained when DNA was isolated from sera using "Proba Rapid" express method. Clinical-epidemiologic diagnosis oftularemia was confirmed by both immune-serologic and RT-PCR methods when sera were studied 3-4 weeks after the onset of the disease.
  
  
  CONCLUSIONS
  RT-PCR with ISFTu2F/R primers and fluorescent probe ISFTu2P, having high sensitivity and specificity, allows to determine F. tularensis DNA in blood sera of patients at both the early stage and 3-4 weeks after the onset of the disease.
authors.forename: M I
authors.initials: MI
authors.lastname: Kormilitsyna
authors.forename: I S
authors.initials: IS
authors.lastname: Mescheryakova
authors.forename: T V
authors.initials: TV
authors.lastname: Mikhailova
authors.forename: A A
authors.initials: AA
authors.lastname: Dobrovolsky
mesh_terms: D004196:Disease Outbreaks; D005604:Francisella tularensis / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D006801:Humans; D007753:Laboratories; D060888:Real-Time Polymerase Chain Reaction; D014406:Tularemia / Q000097:blood* / Q000382:microbiology / Q000473:pathology