REAL TIME POLYMERASE CHAIN REACTION IN TULAREMIA LABORATORY DIAGNOSTICS

• Main Authors: M. I Kormilitsyna, I. S Mescheryakova, T. V Mikhailova, A. A Dobrovolsky
• Format: Article in Journal/Newspaper
• Language: Russian
• Published: Central Research Institute for Epidemiology 2015
• Subjects: francisella tularensis; tularemia laboratory diagnostics; blood sera; dna; real time polymerase chain reaction; primers; probes; Microbiology; QR1-502; khanty
• Online Access: https://doaj.org/article/5338c7e70267404bb2c363950a6ae64d

Aim. Enhancement of tularemia laboratory diagnostics by F. tularensis DNA determination in blood sera of patients using real time polymerase chain reaction (RT-PCR). Materials and 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, Tul4GF/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 Tul4GF/R+tul4-PR2 combination - 92% ofsera. 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. Conclusion. 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.