Molekuliarnaia Genetika, Mikrobiologiia I Virusologiia[JOURNAL]
Sun
200 papers
[Analysis of a GSTM1 gene deletion in the context of the GSTM genomic cluster diversity in three Russian populations].
Filippova IN, Khrunin AV, Limborskaia SA
Mol Gen Mikrobiol Virusol · 2014 · PMID 25080812
A total of 16 to 60% of individuals in human populations are homozygous with respect to a deletion of the Glutathione-S-transferase M1 gene. In this study, we evaluated the relationship between the GSTM1 gene deletion an...
A total of 16 to 60% of individuals in human populations are homozygous with respect to a deletion of the Glutathione-S-transferase M1 gene. In this study, we evaluated the relationship between the GSTM1 gene deletion and genetic diversity of the GSTM cluster, which includes this gene, in three Russian populations. The study was based on the comparison of the haplotype distribution in two groups of individuals subdivided accordingly to the presence of the deletion. The first group included individuals with completely deleted GSTM1 gene, and the second group comprised individuals having at least one functional variant of GSTM1 gene. The analysis of the haplotype frequencies in groups revealed no specificity in their distribution both within the populations and between them.
[Chlamydia trachomatis proteasome protein as one of the significant pathogenicity factors of exciter].
Davydov DIu, Zigangirova NA
Mol Gen Mikrobiol Virusol · 2014 · PMID 25080811
Sex-related infections are a global problem. Such infections may lead to acute or chronic diseases. Chlamydia trachomatis is a dangerous and widespread pathogenicity factor that is not sensitive to conventional drugs and...
Sex-related infections are a global problem. Such infections may lead to acute or chronic diseases. Chlamydia trachomatis is a dangerous and widespread pathogenicity factor that is not sensitive to conventional drugs and has no obvious symptoms. Protein CPAF is leading factor of pathogenesis. This protein inhibits the signaling pathways of host cell and supports long survival of the pathogen in the host cell. The goal of this work was to review general properties of the proteasome Chlamydia protein CPAF, its functions, and role in pathology. The role of protein CPAF in the anti-chlamydia immune reaction is discussed. The prospects of the development of promising anti-chlamydia vaccine, as well as new effective anti-chlamydia drugs are also discussed.
Potassium hydroxide-ethylene diamine tetraacetic acid method for the rapid preparation of small-scale PCR template DNA from actinobacteria.
Sun Z, Huang Y, Wang Y … +2 more , Zhao Y, Cui Z
Mol Gen Mikrobiol Virusol · 2014 · PMID 24757841
Genomic DNA extraction from Gram-positive bacteria is a laborious and time-consuming process. A rapid and convenient method was established to extract genomic DNA from a single colony as a PCR template. KOH-EDTA is used...
Genomic DNA extraction from Gram-positive bacteria is a laborious and time-consuming process. A rapid and convenient method was established to extract genomic DNA from a single colony as a PCR template. KOH-EDTA is used as a lysis buffer to disrupt the cell envelope, releasing genomic DNA, and Tris-HCl (pH = 4) is then added to neutralize the lysate. The lysate can be used directly as a template for PCR amplification. 16S rDNA was successfully amplified from Gram-positive bacteria from the genera of Bacillus, Streptomyces, Micromonospora, Nonomuraea, Microbispora, and Staphylococcus. Amplification of the trpB gene indicated that this method could also be applied to the amplification of functional genes. Compared to colony PCR methods without KOH-EDTA, this method is extremely fast and efficient, and it is applicable to high-throughput PCR amplifications.
[Molecular evolution of the West Nile virus].
Subbotina EL, Loktev VB
Mol Gen Mikrobiol Virusol · 2014 · PMID 24757840
Sixty eight nucleotide sequences encoding protein E of the West Nile virus (WNV) were used for the phylogenetic analysis and estimation of the evolution rate of the WNV. Nucleotide substitution accumulation rate was eval...
Sixty eight nucleotide sequences encoding protein E of the West Nile virus (WNV) were used for the phylogenetic analysis and estimation of the evolution rate of the WNV. Nucleotide substitution accumulation rate was evaluated as 2.5 x 10(-4) substitutions per site per year. Phylogenetic analysis and divergence time estimation carried out using the molecular clocks methodology showed that genotypes 1, 2, and 4 of the WNV circulated in the area of the European Russia with estimated divergence times from a common ancestor of approximately 2360, 2800, and 5950 years ago, respectively. The non-synonymous (dN) to the synonymous (dS) substitution values were found between 0.022-0.275 for the different WNV strains that were grouped by geographical and/or filogenetic characteristics. The highest dN/dS values were found in the group of WNV isolates coming from Russia and North America that have disseminated in these new regions over the past 14 years. Estimation of dN/dS for WNV shows that the dN/ dS value can be used as an indicator of the intraspecies variability and for evaluation of evolution rate for new isolates of WNV. This confirms the hypothesis about of the favorable conditions for the wide dissemination and rapid evolution of different' genotypes of WNV occurring from 2 up to 6 thousand years ago in modern geographical and climatic conditions.
[Genovariants of the cholera agent biovar El Tor: construction, molecular-genetic, and proteomic analysis].
Smirnova NI, Agafonov DA, Shchelkanova EY … +3 more , Zadnova SP, Cherkasov AV, Kutyrev VV
Mol Gen Mikrobiol Virusol · 2014 · PMID 24757839
Experimental modeling of origination of the virulent Vibrio cholerae El Tor genovariants is presented. It was demonstrated that the genovariants obtained did not differ from the natural genetically modified strains emerg...
Experimental modeling of origination of the virulent Vibrio cholerae El Tor genovariants is presented. It was demonstrated that the genovariants obtained did not differ from the natural genetically modified strains emerged in a natural population of the agent, either in phenotypical or genotypic properties. Using the PCR assay and sequencing techniques it was proved that the constructed genovariants carried a CTX(Class phi) prophage genome region with ctxBl gene of the V. cholerae classical biovar in the chromosome. It is shown that the prophage structure alterations lead to the increase in the toxigenicity and virulence in the genovariants compared to the typical strain-recipient. Moreover, as regards proteomics, changes in the expression of 26 proteins that perform various functions in the cell, such as metabolism, energy exchange, transportation, etc., were demonstrated. The data are indicative of the impact that a new DNA region in the genome of the genovariants has on the expression level of different house-keeping genes. The results obtained testify to the fact that one of the mechanisms of the genovariant emergence in the natural populations of the agent can be horizontal gene transfer.
[The manganese-induced infidelity of the DNA synthesis as a possible cause of manganism].
Lakhin AV, Tarantul VZ, Gening LV
Mol Gen Mikrobiol Virusol · 2014 · PMID 24757838
The impact of the 8 most common bivalent metal cations (Mg2+, Mn2+, Co2+, Cd2+, Zn2+, Ni2+, Ca2+, Cu2+) on the operation of the whole complex of DNA polymerases in mice brain cell extracts was tested. A decrease in the f...
The impact of the 8 most common bivalent metal cations (Mg2+, Mn2+, Co2+, Cd2+, Zn2+, Ni2+, Ca2+, Cu2+) on the operation of the whole complex of DNA polymerases in mice brain cell extracts was tested. A decrease in the fidelity of the DNA synthesis was observed in the presence of several metals; among them, Mn2+ caused the most significant effect. It was also demonstrated that this effect was mainly due to the DNA polymerase iota (Pol iota) activity. It is well known that occupational or environmental exposure to excessive Mn could lead to development of neurodegenerative diseases (e.g., manganism). However, the molecular mechanism underlying these pathologies is still unknown. Our results suggest that the neurotoxic effect of Mn2+ may be associated with local activation of highly error-prone Pol iota that increases incorrect DNA synthesis at elevated concentrations of this metal.
[Francisella tularensis strains molecular typing using the multiple-locus variable-number tandem repeat analysis].
Tomofeev VS, Kudryavtseva TY, Mokrievich AN … +2 more , Pavlov VM, Dyatlov IA
Mol Gen Mikrobiol Virusol · 2014 · PMID 24757837 · Publisher ↗
Variable-number tandem repeat analysis (VNTR) of 25 loci was used for molecular typing of the Francisella tularensis strains isolated from different regions of Russia and the former Soviet Union. This approach allowed us...
Variable-number tandem repeat analysis (VNTR) of 25 loci was used for molecular typing of the Francisella tularensis strains isolated from different regions of Russia and the former Soviet Union. This approach allowed us to subdivide F. tularensis subspecies and determine genotype diversity with regard to the geographical prevalence. All 25 loci were examined for their ability to discriminate subspecies and local geographical group. 42 genotypes among the 58 investigated F. tularensis subsp. holarctica isolates were found using cluster VNTR analysis.
[A study of the single nucleotide polymorphism fragments of the AUR gene metalloprotease strains of Staphylococcus aureus isolated from the skin of patients with atopic dermatitis].
Tuyrin YA, Shamsutdinov AF, Fassahov RS
Mol Gen Mikrobiol Virusol · 2014 · PMID 24757836
OBJECTIVE: to study the heterogeneity of internal fragments of the aur gene coding a proteolytic enzyme aureolysin in strains of Staphylococcus aureus with different enzymatic activity, which were allocated with the affe...
OBJECTIVE: to study the heterogeneity of internal fragments of the aur gene coding a proteolytic enzyme aureolysin in strains of Staphylococcus aureus with different enzymatic activity, which were allocated with the affected skin of patients with atopic dermatitis. MATERIALS AND METHODS: the study included 125 patients aged 1 to 35 years old with the diagnosis of atopic dermatitis. 100 clinical strains of Staphylococcus aureus were studied in monoculture. Identification of the strains of Staphylococcus aureus in skin was performed using standard classical methods. Specific biochemical microtests using a set of APIs Staph 20 (bioMerieux SA, France) were conducted to confirm the obtained results. Reference strains of S. aureus ATCC 29213, NCTC at 8.325 were used. Specific aur gene loci were detected by the method of multiplex polymerase chain reaction. Proteolytic activity was determined by the ability to split the substrate (human IgG, Sigma) into fragments using as inhibitors of matrix 0.01-0.1M solutions of sodium EDTA. The sequencing of the amplified areas of aur gene, including the preliminary stages of separation and purification of DNA, was performed by Syntol, Moscow. RESULTS: different strains of the conservative plot amplicon fragment of aur contains up to 9 polymorphic nucleotides. The index of the nucleotide diversity for proteolytic active strains was less (Pi = 0.04) than for proteolytically inactive (Pi = 0.07), p < 0.05. It should be noted that all proteolytically active strains were isolated from the skin of patients with high and moderate severity of atopic dermatitis (67.8 +/- 5.4 points on SCORAD scale) with erythematous form of the disease. The inactive strains were allocated in patients of older age with reduced severity of atopic dermatitis (with the dry form of skin lesions).
[Analysis of mutations in patients with suspected autosomal dominant form of the Parkinson disease].
Filatova EV, Alieva AKh, Shadrina MI … +5 more , Shulskaya MV, Fedotova EY, Illarioshkin SN, Limborska SA, Slominsky PA
Mol Gen Mikrobiol Virusol · 2014 · PMID 24757835
The Parkinson disease (PD) is a severe neurological disorder. Diverse genetic systems and environmental factors are involved in the pathogenesis of this disease. However, despite extensive research into the disease, its...
The Parkinson disease (PD) is a severe neurological disorder. Diverse genetic systems and environmental factors are involved in the pathogenesis of this disease. However, despite extensive research into the disease, its causes are not fully elucidated, and the exact spectrum of genes and mutations involved in the development of hereditary forms of PD has not been fully clarified yet. The present work is devoted to the analysis of mutations that lead to the development of monogenic forms of PD in patients with suspected autosomal dominant form of PD using Multiplex Ligation-dependent Probe Amplification (MLPA). We have identified several mutations (G2019S in LRRK2, heterozygous deletions of 2-3, 3-4 exons and heterozygous duplication of 2-4 exons in PARK2, deletion of 3 exon in PARK7) that lead to the development of PD in only 7 people out of 70 (18.4%), which suggests the need for further search of new mutations, for example, using exome sequencing. In the future it will help to develop the molecular genetic tests for early preclinical diagnosis and risk evaluation of the development of PD, and to understand better the causes and mechanisms of this disease.
[Genotypic identification of the bovine leukemia virus].
Vafin RR, Khazipov NZ, Shaeva AIu … +5 more , Zakirova ZR, Zaĭnullin LI, Tiul'kin SV, Abdulina IR, Alimov AM
Mol Gen Mikrobiol Virusol · 2014 · PMID 25845140
Phylogenetic analysis of the sequenced segments of the provirus BLV locus env-gene and the strategy of the PCR-PDRF-genotyping consistent with phylogenetic classification of pathogenic agent and suggested in our works pr...
Phylogenetic analysis of the sequenced segments of the provirus BLV locus env-gene and the strategy of the PCR-PDRF-genotyping consistent with phylogenetic classification of pathogenic agent and suggested in our works provided taxonomic identification of BLV isolates identified in cattle in Tatarstan (Russian Federation) as representatives of the 4th, 7th, and 8th BLV genotypes. Of 100 identified isolates, 64 represent the 4th BLV genotype, 28 representatives of BLV belong to cluster of the 7th genotype, whereas the other 8 samples of the provirus belong to the new 8th genotype of pathologic agent. The strategy VBL PCR-PDRF-genotyping suggested in our work on the basis of 5 restriction endonucleases (PvuII, SspI, HphI, HaeIII, and BstYI) provided correct genotyping identification of the viral pathogen.
[Molecular-genetic analysis of the genomes of porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 circulating in the area of Russian Federation].
Bulgakov AD, Grebennikova TV, Iuzhakov AG … +2 more , Aliper TI, Nepoklonov EA
Mol Gen Mikrobiol Virusol · 2014 · PMID 25845139
The molecular genetic analysis of the genomes of the virus of porcine reproductive respiratory syndrome (VPRRS) and porcine circovirus type 2 (PCV-2) circulating in the area of the Russian Federation was discussed. The r...
The molecular genetic analysis of the genomes of the virus of porcine reproductive respiratory syndrome (VPRRS) and porcine circovirus type 2 (PCV-2) circulating in the area of the Russian Federation was discussed. The results of this work showed the circulation of the strains of the European genotype VPRRS similar to those found in France and Denmark from 1998 to 2001. The homology of the fragment of one of the genes between the Russian isolates and the vaccine strain Porcilis PRRS (Intervet) was found. It requires further study. The strains representing the North American genotype VPRRS were not found. The PCV-2 genomes fall into three separate goups. One (genotype 2b) is formed by isolates in Malaysia, Brazil, Switzerland, China, Slovakia, UK, USA, isolated during the period from 2004 to the present time. The second group consists of sequences of the viruses isolated in 2000-2012 in Canada, the U.S., China, and South Korea (genotype 2a). The third group is formed by highly pathogenic isolates in 2013 from China (highly pathogenic genotype 2c). The circulation of all three known genotypes of PCV-2: 2a, 2b, and 2c in Russian Federation was demonstrated.
[Crimean-Congo hemorrhagic fever virus in Stavropol region in 2011].
Iashina LN, Malyshev BS, Netesova NA … +2 more , Volynkina AS, Vasilenko NF
Mol Gen Mikrobiol Virusol · 2014 · PMID 25845138
The genetic analysis of the Crimean-Congo hemorrhagic fever (CCHF) virus circulating in Stavropol region during 2011 year was suggested. A total of 14 RNA isolates from the Crimean hemorrhagic fever patients were genetic...
The genetic analysis of the Crimean-Congo hemorrhagic fever (CCHF) virus circulating in Stavropol region during 2011 year was suggested. A total of 14 RNA isolates from the Crimean hemorrhagic fever patients were genetically typed. The genetic analysis of the CCHF virus stains based on M-segment sequences (positions 2607-2932) supported the circulation of the genotype Europe 1 in the Stavropol region of Russia. In addition to previously known lineage STV-ROS, the second lineage VLG/ROS was observed in Stavropol region.
[A single-tube real-time RT-PCR assay for the RNA detection and quantification of genetically diverse HIV Including rare non-M group of the HIV-1 and HIV-2].
Prasolova MA, Ivanov MK, Gashnikova NM … +2 more , Netesova ES, Dymshyts GM
Mol Gen Mikrobiol Virusol · 2014 · PMID 25845137
The RT-PCR method with real-time fluorescence detection was used for development of φ prototype of diagnostic kit for reliable diagnosis of genetic variants of RNA of the HIV-1 of groups M, N, O, P and RNA of the HIV-2 i...
The RT-PCR method with real-time fluorescence detection was used for development of φ prototype of diagnostic kit for reliable diagnosis of genetic variants of RNA of the HIV-1 of groups M, N, O, P and RNA of the HIV-2 in blood plasma and serum. The kit is stable against nucleotide defects, provides broad linear range of concentration of the HIV RNA, 100% analytical specificity and adequate analytical sensitivity: 42 ME/ml (HIV-1 of group M), 45 copies/ml (HIV-2), 92 copies/ml (HIV-1 of group O), 90 copies/ml (HIV-1 of group N). The kit provided successful detection and measurement of HIV RNA concentration in the samples of the international reference panel of the HIV-1 genotype. The results of the test correlate with results of commercial tests.
[The genome polymorphism of the Mycobacterium avium subsp. hominissuis strains].
Starkova DA, Mokrousov IV, Viazovaia AA … +4 more , Zhuravlev VIu, Otten TF, Vishnevskiĭ BI, Narvskaia OV
Mol Gen Mikrobiol Virusol · 2014 · PMID 25845136
The non-tuberculosis mycobacteria Mycobacterium avium subsp. hominissuis (MAH) are able to cause human mycobacteriosis. In this work, the results of the first comprehensive study of the genome polymorphism of the clinica...
The non-tuberculosis mycobacteria Mycobacterium avium subsp. hominissuis (MAH) are able to cause human mycobacteriosis. In this work, the results of the first comprehensive study of the genome polymorphism of the clinical strains of MAH were reported using the typing scheme by 13 loci MATR-VNTR (TR292, TRX3, TR25, TR47, MATR-1, MATR-4, MATR-5, MATR-6, MATR-8, MATR-11, MATR-14, MATR-15, MATR-16) containing tandem nucleotide sites and IS1245-RFLP-typing sites. A total of 90 MAH strains isolated from patients with lung mycobacteriosis without epidemiological connection (including HIV infected) were tested in 2008-2011. The inhomogeneity of the MAH strains by 36 profiles of 13 loci MATR-VNTR was observed. The majority of the strains (68.8%) were included in the 8 MATR-VNTR clusters; most large cluster contained 37 strains with 13-bitnumerical profile 2222223145443'. The nucleotide sequence of the MATR-16 (3') locus contains the long deletion (GenBank accession no. KF479191). The MAH strains of the MATR-VNTR clusters were found to be inhomogeneous by the IS1245 marker. The MATR-VNTR-typing method by 13 loci is recommended for preliminary differentiation of domestic MAH strains with further analysis of the MATR-VNTR clusters using the IS1245-RFLP-typing method.
[The ability of the natural ketones to interact with bacterial quorum sensing systems].
Pliuta VA, Popova FF, Koksharova OA … +2 more , Kuznetsov AE, khmel' IA
Mol Gen Mikrobiol Virusol · 2014 · PMID 25845135
The effect of the natural ketones emitted by bacteria (2-nonanone, 2-heptanone, 2-undecanone) on the functioning of the Quorum Sensing (QS) systems was studied. In this work, three lux-reporter strains containing the com...
The effect of the natural ketones emitted by bacteria (2-nonanone, 2-heptanone, 2-undecanone) on the functioning of the Quorum Sensing (QS) systems was studied. In this work, three lux-reporter strains containing the components of the LasI/LasR, RhlI/RhlR, LuxI LuxR QS systems were used as biosensors for the N-acyl-homoserine lactones. It was shown that at concentrations of ketones that exhibited little or no bactericidal action the ketones could modulate the QS-response by suppressing the expression of the lux-operon reporter to a greater extent than the cell viability of these strains.
[Codon optimization of hemagglutinin as a promising tool for improving of the influenza vaccines immunogenicity].
Fedorova EF, Kiseleva IV, Auewarakul P … +2 more , Suptawiwat O, Rudenko LG
Mol Gen Mikrobiol Virusol · 2014 · PMID 25845134
Modification of the codon bias of sequences is a promising tool of the gene expression control. The theoretical basis of the codon optimization is reviewed, data on experiments in changing the viral gene codon bias for p...
Modification of the codon bias of sequences is a promising tool of the gene expression control. The theoretical basis of the codon optimization is reviewed, data on experiments in changing the viral gene codon bias for purposes of vaccine development are discussed. Research into the field of the influenza vaccine immunogenicity improvement with codon optimization method is reviewed. Prospects of the use of the codon optimization technique for influenza vaccine development are considered.
PCR-ELISA: a diagnostic assay for identifying Iranian HIV seropositives.
Bagheri R, Rabbani B, Mahdieh N … +3 more , Khanahmad H, Abachi M, Asgari S
Mol Gen Mikrobiol Virusol · 2013 · PMID 24364144
Quantitative viral load monitoring is an important indicator of prognosis in human immunodeficiency virus type 1 (HIV-1). PCR-ELISA as a quantitative method is proven to be sensitive and specific for quantification of HI...
Quantitative viral load monitoring is an important indicator of prognosis in human immunodeficiency virus type 1 (HIV-1). PCR-ELISA as a quantitative method is proven to be sensitive and specific for quantification of HIV-1. Extracted DNA of thirty seropositive and twenty seronegative individuals which were confirmed by ELISA and western blot were amplified with digoxigenin- labeled nucleotides; and then in ELISA procedure biotin-labeled probes were hybridized to the PCR products. Diluted PCR products were analysed by electrophoresis and ELISA methods. The observation revealed that combination of nested-PCR and ELISA leads to a sensitive and specific identification of three copies in HIV-infected; the specificity and sensitivity were 95% and 96.7%, respectively. In conclusion, PCR-ELISA was 10 fold more sensitive than nested-PCR. This study developed a high sensitive PCR-ELISA to assess the quantification of proviral DNA load in the most relevant case of HIV-1 subtype. The reproducibility and reliability of this high-throughput test makes it appropriate for general laboratories to use for quantification of viral load and clinical diagnosis.
[Genetic variants of the Crimean-Congo hemorrhagic fever virus circulating in endemic areas of the southern Tajikistan in 2009].
Petrova ID, Kononova IuV, Chausov EV … +2 more , Shestopalov AM, Tishkova FKh
Mol Gen Mikrobiol Virusol · 2013 · PMID 24364143
506 Hyalomma anatolicum ticks were collected and assayed in two Crimean-Congo hemorrhagic fever (CCHF) endemic regions of Tajikistan. Antigen and RNA of CCHF virus were detected in 3.4% of tick pools from Rudaki district...
506 Hyalomma anatolicum ticks were collected and assayed in two Crimean-Congo hemorrhagic fever (CCHF) endemic regions of Tajikistan. Antigen and RNA of CCHF virus were detected in 3.4% of tick pools from Rudaki district using ELISA and RT-PCR tests. As of Tursunzade district, viral antigen was identified in 9.0% of samples and viral RNA was identified in 8.1% of samples. The multiple alignment of the obtained nucleotide sequences of CCHF virus genome S-segment 287-nt region (996-1282) and multiple alignment of deduced amino acid sequences of the samples, carried out to compare with CCHF virus strains from the GenBank database, as well as phylogenetic analysis, enabled us to conclude that Asia 1 and Asia 2 genotypes of CCHF virus are circulating in Tajikistan. It is important to note that the genotype Asia 1 virus was detected for the first time in Tajikistan.
[Molecular genetic analysis of the isoniazid-resistant strains of M. tuberculosis circulating over the Saratov region].
Salina TIu, Morozova TI
Mol Gen Mikrobiol Virusol · 2013 · PMID 24364142
257 patients with active lung tuberculosis in the Saratov region were tested for distribution and mutation spectrum of the genes katG, inhA, ahpC encoding Isoniazid-resistance. The tests were implemented using the biolog...
257 patients with active lung tuberculosis in the Saratov region were tested for distribution and mutation spectrum of the genes katG, inhA, ahpC encoding Isoniazid-resistance. The tests were implemented using the biological microchips with pharmacological kits TV-Biochip MDR (Russia). The rate of the Isoniazid-resistant M. tuberculosis strains incidence was found to be 55.7%. The Isoniazid-resistant strains demonstrated high rate (26.9%) of combined mutations in 2 and 3 genes simultaneously, domination in the katG ser315 -> Thrl (91.1%), thereby inducing high resistance to Isoniazid. The sensitivity of the TV-Biochip MDR test-system was 88.2%; the specificity, 91.3%. The results ofthis work show that the incidence of the Isoniazid-resistant strains should be limited using additional measures.
[The molecular and genetic characterization of Francisella tularensis strains of different taxonomic status and virulence].
Kormilitsyna MI, Meshcheriakova IS, Mikhaĭlova TV
Mol Gen Mikrobiol Virusol · 2013 · PMID 24364141
Typing of Francisella strains collection by means of PCR on the basis of tul4 and RDI genes was carried out. The identification of the species and subspecies of the 112 strains of Francisella tularensis was reashed. The...
Typing of Francisella strains collection by means of PCR on the basis of tul4 and RDI genes was carried out. The identification of the species and subspecies of the 112 strains of Francisella tularensis was reashed. The PCR on DNA-targets loci of type IV pili genes: pilA, pilE2, pilE3, pilE4, pilE5, pilF, pilT, pilD and pilQ for differentiation of F. tularensis strains on virulence was carried out. It was demonstrated the possibility of differentiation of F. tularensis strains in PCR (primers A-B) on the basis of the revelation of the gene pilA in virulent strains of third subspecies F. tularensis and F. tularensis subsp. novicida. This gene pilA was not detected in the vaccine strain 15/10 and its variants, as well as in the most of avirulent F. tularensis subsp. holarctica strains. However, the fragment gene pilA was found in the attenuated strains F. tularensis subsp. tularensis and mediasiatica. It was not revealed any differences on other targets of pili genes between of F. tularensis strains, with the exception of the strain F. tularensis subsp. novicida Utah 112, which had not a fragment of the gene pilE2. The use of PCR to target the locus of the pilA gene allows to discriminate virulent F. tularensis subsp. holarctica strains from the vaccine 15/10, its variants and avirulent strains.
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