Zhurnal mikrobiologii, epidemiologii i immunobiologii · 2013
[New approaches to detection of legionellosis causative agents in clinical samples and environment objects by PCR].
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Portenko S A, Osina N A, Bugorkova T V, Kutyrev V V
Аннотация
AIM: Selection of perspective targets as a base for development of test systems for the detection of legionella DNA in study material by PCR with electrophoresis and hybridization-fluorescent accounting of the results.
MATERIALS AND METHODS: 22 Legionella pneumophila, 3 Legionella spp. strains and 30 cultures ofheterologic microorganisms, clinical material and environmental object samples were used in the study. Genome analysis was carried out by using Mega 3.1 program. Primer selection was conducted by using Primer Express program and BLAST algorithm and TaqMan format probes on the website www.genscript.com.
RESULTS: Analysis of 712 legionella nucleotide sequences for the presence of novel species-specific and conservative for L. pneumophila loci was carried out. Fragments of life-support genes were selected for the analysis: fliC, mompS, ftsZ, dotA, dnaX, trpS, rpoB, rnp, proA, gspA. The most perspective DNA targets were established to be ftsZ and mompS genes. Based on the selected loci, PCR test systems were constructed for the detection of DNA oflegionella causative agent in biological material and environment objects, their diagnostic value was characterized.
CONCLUSION: The studies carried out have shown the perspective of use of life-support ftsZ and mompS genes for the construction of novel preparations for legionellosis genetic diagnostics.
MATERIALS AND METHODS: 22 Legionella pneumophila, 3 Legionella spp. strains and 30 cultures ofheterologic microorganisms, clinical material and environmental object samples were used in the study. Genome analysis was carried out by using Mega 3.1 program. Primer selection was conducted by using Primer Express program and BLAST algorithm and TaqMan format probes on the website www.genscript.com.
RESULTS: Analysis of 712 legionella nucleotide sequences for the presence of novel species-specific and conservative for L. pneumophila loci was carried out. Fragments of life-support genes were selected for the analysis: fliC, mompS, ftsZ, dotA, dnaX, trpS, rpoB, rnp, proA, gspA. The most perspective DNA targets were established to be ftsZ and mompS genes. Based on the selected loci, PCR test systems were constructed for the detection of DNA oflegionella causative agent in biological material and environment objects, their diagnostic value was characterized.
CONCLUSION: The studies carried out have shown the perspective of use of life-support ftsZ and mompS genes for the construction of novel preparations for legionellosis genetic diagnostics.