REAL TIME POLYMERASE CHAIN REACTION IN TULAREMIA LABORATORY DIAGNOSTICS

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 201...

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Bibliographic Details
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:
dna
Online Access:https://doaj.org/article/5338c7e70267404bb2c363950a6ae64d
Description
Summary: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.