PubMed Health⌕ Search

Biomedical subjects

A N Kulichenko

Publications and source records attributed to A N Kulichenko.

17 recordsLinked to original sources
PubMed2005

[Laboratory diagnostics of urogenital clamidiosis].

Laboratory diagnostic tools of urogenital clamidiosis--PCR, ELISA (IgG and IgM) and direct immunofluorescence (DIF)--were comparatively analyzed. The positive PCR result was checked by another PCR test with a different primer; 5 false positive responses were registered (specificity 99.6%). As against PCR, the sensitivity of ELISA made 53%, its specificity -75.5%, the diagnostic value of positive result -58%, the diagnostic value of negative result -71.6% and the diagnostics accuracy -66.7%. The respective DIF parameters were as follows: 36%, 90%, 81.5%, 54.2% and 60.9%. A high rate of detection (above 90%) of the conditionally pathogenic microflora associated with Chlamydia trachomatis (above 110 microbe cells/ml) was pointed out. Hardnerelli and ureaplasms were more often found in female smears, staphylococci and enterococci--in male sperm. It is underlined as important to hold complex examinations for Chlamidia (PCR, ELISA and DIC as an additional test) combined with bacteriological quantification of the conditionally pathogenic microflora and determination of its resistance to antibiotics.

Chlamydia Infections↗
PubMed2004

[Paraendocervical route for metronidazole administration in complex treatment of chronic Trichomonas endocervicitis].

The data on paraendocervical administration of metronidazole in the treatment of patients with relapsing Trichomonas endocervicitis are presented. Metronidazole was administered as 0.5% solution in a dose of 0.04 g once a day for 8-10 days in complex traditional therapy including oral use of metronidazole and immunocorrigating and local treatment. It was shown that paraendocervical administration of the protistocidal agent provided earlier regression of the urogenital symptoms and 2.8 times lower frequency of the relapses.

Administration, Intravaginal↗
PubMed1994

Development of a diagnostic test for Yersinia pestis by the polymerase chain reaction.

A 501 bp caf1 gene fragment and a 443 bp of pla gene fragment carried by 100 kb (pFra) and 10 kb (pPst) species-specific extrachromosomal replicons, respectively, were used as targets to study the conditions under which DNA amplification by polymerase chain reaction (PCR) may be applied to detect and identify Yersinia pestis DNA in cell lysates of pure cultures and biological samples. The sensitivity limit of PCR with the crude cell lysates of Y. pestis EV was estimated as 10-50 cfu in reaction mixture. When target Y. pestis EV cells were mixed with fresh blood of white mice, which contained 0.4% potassium citrate, the PCR detection level varied from 400 to 100 cfu ml-1 of blood depending on the method used for preparing the sample. In our tests PCR was effective for the detection of yersinia in the blood of white laboratory mice experimentally infected with virulent Y. pestis KM638 strain. This method can be considered convenient for routine detection and identification of Y. pestis.

Animals↗
PubMed1994

[Improvement of a method for detecting of strains of the plague microbe using polymerase chain reaction].

Three pairs of oligonucleotide primers, complementary to nucleotide sequences of Yersinia pestis plasmids (pPst, 9.5 kb; pCad, 70 kb; pFra, 95 kb), were used in polymerase chain reaction for high-sensitive specific detection of plague pathogen. Primer pairs P1,P2,C1,C2, and F1,F2 were used to amplify fragments of pla gene (plasmid pPst), yop1 gene (plasmid pCad of plague microbe), and caf1 gene (plasmid pFra), respectively. The method developed enables specific detection of strains from all natural loci in Russia and contiguous states as well as strains of oceanic origin. Sensitivity of method is 50-100 CFU/ml. Primer sequences enable to amplify gene fragments, located in three own plasmides of plague microbe, in the same reaction mixture. The method offers identification of plague microbe and determination of its virulence and epidemic significance.

Base Sequence↗
PubMed1993

[Design of a chromosomal DNA probe species specific for Yersinia pestis].

A 14.8 kb DNA fragment from the chromosome of Yersinia pestis TWJ was cloned and the restriction map constructed. The fragment designated as T16 and its subfragments were tested in dot-hybridization with strains of Yersinia genus and other members of Enterobacteriaceae. A species-specific DNA probe (designated MK) was constructed on the basis of the T16 fragment. As judged from restriction analysis, blot-hybridization experiments and, partially, sequencing, significant homology exists between the MK DNA probe and this one developed by Bardarov et. al. (1990). A repeated sequence in two copies was discovered in the MK fragment.

Base Sequence↗
PubMed1991

[Constructing a test system for the identification of plague microbe based on genetic probes].

DNA probes for detection of the plague agent Yersinia pestis were made on a basis of its three typical extrachromosomal replicons. The recombinant plasmid pBS2 including pBR327 vector and SalGI-BspRI fragment of the plasmid pFra was constructed. The above fragment is connected with synthesis of Y. pestis capsular antigen and it is a 400 bp species-specific DNA probe called F1 which is suitable for identification of Y. pestis species that bears the 60 mdal plasmid. The DNA probes called P1 was made on a basis of the plasmid pPst; it is the 460 BglII-BamHI fragment of the fibrinolysin-coagulase gene suitable for species-specific detection of Y. pestis species that bears the 60 mdal plasmid. The P1 fragment was cloned into the pAT153 vector and the constructed recombinant plasmid was called pEK7. The recombinant plasmid pCL1, including the pBR325 vector and the 6th BamHI fragment of Y. pestis EV plasmid pCad was constructed. The above fragment includes the replication origin of the pCad and it is hybridized to the pCad-bearing strains of Y. pestis and Y. tuberculosis only. Thus, it may be a basis for a bi-species-specific DNA probe making. These three recombinant plasmids are considered as a test-system for detection of both typical and atypical strains of Y. pestis.

Bacteriological Techniques↗
PubMed1989

[The development of a method of nitrosoguanidine inactivation in the mutagenic treatment of bacteria].

A simple method for the inactivation of nitrosoguanidine, a mutagenic agent permitting the simplification and acceleration of the procedure used for obtaining bacterial mutants, is described. This method is based on the addition of unithiol to nitrosoguanidine solution to a final concentration of 0.25-0.5% and the subsequent normalization of the pH of the medium with 0.1 N NaOH solution.

Bacteria↗
PubMed1988

[A new pathogenic trait encoded by Yersinia pseudotuberculosis pVM82 plasmid].

The strains of Yersinia pseudotuberculosis isolated from patients in the course of outbreaks of infection (epidemic strains) were found to possess at least two plasmids with molecular masses of 45 and 82 MD. In contrast, the strains obtained in sporadic cases harbored different sets of plasmids, but never the 82 MD plasmids. These plasmids designated pVM82 and isolated from strains of different geographic regions of the country were identical. pVM82 have no homology with Y. pestis plasmids of the similar size coding for the FraI antigen. The pVM82 DNA was found to be composed of the 57 MD plasmid DNA and the 25 MD fragment of Y. pseudotuberculosis DNA. Using Western blot hybridization technique it was shown that the presence of pVM82 suppressed formation of antibody against some major antigenic determinants of Y. pseudotuberculosis. Immunosuppression took place when the animals were infected with bacteria grown below 20 but not at 37 degrees C. The 57 MD plasmid failed to produce immunosuppression. It was concluded that the 25 MD fragment of pFN82 encoded a novel pathogenic factor responsible for immunosuppression.

Antibodies, Bacterial↗
PubMedDate not supplied

[Characterization of Vibrio cholerae eltor isolates according to their epidemic potential using new diagnostic cholera bacteriophages eltor ctx+ and ctx- and by the polymerase chain reaction].

The epidemic potential of 113 V. cholerae eltor strains of different origin was determined with new diagnostic cholera bacteriophages eltor ctx+ and ctx-, as well as the test for hemolytic activity. Of these strains 50 were epidemically safe and 51 were epidemically dangerous, while the epidemic potential of 12 other strains could not be detected. Determination of genes ctxA, tcpA and toxR in the strains under study by means of the polymerase chain reaction (PCR) revealed that epidemically dangerous strains carried the whole set of the above genes in 92.2% of cases. 98.0% of epidemically safe cultures were lacking either gene ctxA, or genes ctxA and tcpA, or genes ctxA, tcpA and toxR, which confirmed their incapacity to cause cholera. The results of the differentiation of the cultures with new diagnostic cholera phages coincided with the results of PCR in 90% of cases. The most complete and reliable evaluation of the epidemic potential of individual vibrio isolates may be obtained using the two compared methods. The amplification test system gives more information when isolates with unclear epidemic potential are analyzed.

Bacterial Outer Membrane Proteins↗
PubMedDate not supplied

[Anti-arbovirus antibodies positive serum screening in population of Saratov].

Seroepidemiological studies revealed the existence of a population stratum immune to West Nile and Sindbis viruses in the Saratov region. Screening of 218 sera showed that the stratum immune to arboviruses increased from 1998 to 2000. The incidence of antibodies to Batai and Sindbis viruses was the highest. Activation of natural foci of these arboviruses can lead to exacerbation of the epidemic situation in the region.

Antibodies, Viral↗
PubMedDate not supplied

[Comparative assessment of the diagnostic value of the laboratory diagnostic methods for trichomoniasis].

The authors compared the sensitivity and specificity of currently available methods for laboratory diagnosis of trichomoniasis, by examining 971 persons. The highest frequency of T. vaginalis was detected by studies of a stained smear (37.4%), culture tests (19.0%); polymerase chain reaction (PCR) (17.1%), immunofluorescence tests (12.7%), wet smear test (2.7%). Enzyme immunoassay yielded positive results in 36.2% of cases. The use of PCR and culture tests frequently provided similar results (92.0%). The authors have proposed an algorithm of a laboratory study including wet smear microscopy, PCR, and culture tests as basic methods. They have noted the higher detection rates of T. vaginalis when analyzing during draining physiotherapeutic procedures (endocervical vibratory massage with vacuum aspiration in females and pneumovibratory massage of the prostate with endourethral chemotrypsin electrophoresis in males).

Animals↗
PubMedDate not supplied

[The ecology and prevalence of arboviruses on the territory of the Saratov Region].

The territorial spread of Tahyna, Batai, Sindbis, West Nile fever and Crimean-Congo hemorrhagic fever viruses throughout the Saratov region in 1998 - 2000 was analyzed. The characteristics of the epizootic activity of the natural foci of these arboviruses in different landscape zones (temperate forest-steppes, steppes and semi-deserts) were calculated. The species composition of small mammals, the natural reservoirs of the causative agents of arbovirus infections, was determined.

Animals↗
PubMedDate not supplied

[A gene expression study and the isolation and immunobiological properties of the iron-regulated membrane proteins of the plague microbe].

The study has examined how the plague bacillus expresses genes of two high-molecular weight (190 and 240 kD) iron-regulated membrane proteins (HMWP). The latter were purified and assayed for their immunobiological properties in vitro and in vivo. DNA-probing (at E.Carniel's Laboratory, Pasteur Institute, Paris) has indicated that Yersinia pestis strains having genes that code for HMWP synthesis circulate in the natural foci of plague which are situated in the former USSR. The use of HMWP in combination with live plague vaccine EV was found to greatly enhance its immunogenicity.

Animals↗
PubMedDate not supplied

[A DNA analysis of Vibrio cholerae strains by the polymerase chain reaction].

The method for the analysis of cholera toxin gene in V. cholerae strains was developed on the basis of polymerase chain reaction (PCR). This specific and highly sensitive method using primers affecting the site of the DNA of the operon of cholera toxin gene made it possible to identify one copy of V. cholerae genome. For the first time the content of cholera toxin gene in 4 V. cholerae (eltor) strains, obtained from the clinical material of cholera patients in Tajikistan and Dagestan, was shown with the use of PCR.

Base Sequence↗
PubMedDate not supplied

[The determination of the content of the cholera toxin gene in the composition of the DNA from Vibrio cholerae strains by means of the nested polymerase chain reaction].

The possibility of detecting cholera toxin genes in V.cholerae enterotoxigenic strains by the method of "nested" polymerase chain reaction with the use of primers on the DNA area of operon ctx of AB genes. The possibility of the detection of several V.cholerae cells by this method was shown with the use of a series of bacterial lysate dilutions. The newly developed test system for the detection of cholera toxin gene on the basis of the analysis of bacterial lysates of V.cholerae nontoxigenic strains, as well as Escherichia coli toxigenic and nontoxigenic strains, was shown to be highly specific.

Base Sequence↗
PubMedDate not supplied

[Molecular genetic methods for detecting and identifying the plague microbe].

The review analyzes the data on the new designed drugs and ways of identification and detection of the plague causative agent. It gives the results obtained from molecular genetic studies in designing the diagnostic test system based on DNA [correction of DNK] probes and in developing a way of detecting the plague causative agent by the polymerase chain reaction.

Animals↗