ABSTRACT
AIM: Comparison of diagnostic capabilities of 2 variants of PCR for detection of Coxiella burneli persistence in dynamics of infectious process in patients with Q fever. MATERIALS AND METHODS: 110 samples of clinical material, obtained from patients with Q fever in an endemic region for this infection (Astrakhan region), were studied. The samples were studied in a standard PCR (marker - 16S rRNA gene fragment) and in real-time. PCR (RT-PCR) (marker - groEL gene fragment). RESULTS: Both markers were established to be perspective for detection of C. burnetii DNA in clinical material, and RT-PCR detects positive result including late stages of the disease (illness day 21 - 31). CONCLUSION: This study is the first Russian publication on comparison on different PCR variants for detection of C. burnetii in blood of Q fever patients in dynamics of the infectious process.
Subject(s)
Bacterial Proteins/genetics , Chaperonin 60/genetics , Coxiella burnetii/genetics , Q Fever , RNA, Bacterial/genetics , RNA, Ribosomal, 16S/genetics , Real-Time Polymerase Chain Reaction , Adult , Female , Humans , Male , Middle Aged , Q Fever/diagnosis , Q Fever/geneticsABSTRACT
We analyzed restriction fragment length polymorphism of lipL32 gene encoding outer membrane protein in three Leptospira genomic species (L. interrogans, L. kirschneri, and L. borgpetersenii) using Bsa29 I and Bam HI restriction endonucleases. It was found that restriction profiles of the studied gene were similar at both the intraspecific and serogroup levels; variability was revealed only at the interspecific level, which can be explained by relatively low variability of lipL32 gene.
Subject(s)
Bacterial Outer Membrane Proteins/genetics , Leptospira/genetics , Lipoproteins/genetics , Leptospira interrogans/genetics , Polymorphism, Restriction Fragment LengthABSTRACT
AIM: Improvement of PCR for detection of Coxiella burnetii in field material by development of set of primers for amplification of groEL gene fragment. MATERIALS AND METHODS: C. burnetii strains, samples from organs of wild rodents and laboratory animals, blood of healthy donors and laboratory animals. Methods of DNA isolation, PCR, and electrophoresis in agarose gel were used. RESULTS: Nucleotide sequences of primers amplifying groEL gene fragment, which could increase specificity of PCR during work with field material were proposed. CONCLUSION: Obtained data open perspective for using this method for detection of C. burnetii in environment.
