Genomic Analysis of Corynebacterium diphtheriae Strains Isolated in the Years 2007-2022 with a Report on the Identification of the First Non-Toxigenic tox Gene-Bearing Strain in Poland.

Infections caused by non-toxigenic Corynebacterium diphtheriae have been reported every year in Poland since 2004, with the ST8 biovar gravis strains being most commonly isolated. This study analyzed thirty strains isolated between 2017 and 2022 and six previously isolated strains. All the strains were characterized using classic methods in terms of species, biovar level, and diphtheria toxin production, as well as by means of whole genome sequencing. The phylogenetic relationship based on SNP analysis was determined. The number of C. diphtheriae infections has been rising in Poland every year with a maximum of 22 cases in the year 2019. Since 2022, only the non-toxigenic gravis ST8 (most common) and mitis ST439 (less common) strains have been isolated. An analysis of the genomes of the ST8 strains showed that they had many potential virulence factors, such as adhesins and iron-uptake systems. The situation rapidly changed in 2022 and strains from different STs were isolated (ST32, 40, and 819). The ST40 biovar mitis strain was found to be non-toxigenic tox gene-bearing (NTTB), with the tox gene inactivated due to a single nucleotide deletion. Such strains were previously isolated in Belarus. The sudden appearance of new C. diphtheriae strains with different STs and the isolation of the first NTTB strain in Poland indicate that C. diphtheriae should be classified as a pathogen of special public health concern.

Effectiveness of experimental and commercial pertussis vaccines in the elimination of Bordetella pertussis isolates with different genetic profiles in murine model.

The aim of this study was to compare the elimination of Bordetella pertussis clinical isolates, representing different genotypes in relation to alleles encoding virulence factors (MLST-multi-locus antigen sequence typing), MLVA type (multi-locus variable-number tandem repeat analysis) and PFGE group (pulsed-field gel electrophoresis) from the lungs of naive mice or mice were immunised with the commercial whole-cell pertussis vaccine, the acellular pertussis vaccine and the experimental whole-cell pertussis vaccine. Molecular data indicate that the resurgence of pertussis in populations with high vaccine coverage is associated with genomic adaptation of B. pertussis, to vaccine selection pressure. Pertactin-negative B. pertussis isolates were suspected to contribute to the reduced vaccine effectiveness. It was shown that one of the isolates used is PRN deficient. The mice were intranasally challenged with bacterial suspension containing approximately 5 × 10 7 CFU/ml B. pertussis. The immunogenicity of the tested vaccines against PT (pertussis toxin), PRN (pertactin), FHA (filamentous haemagglutinin) and FIM (fimbriae types 2 and 3) was examined. The commercial whole-cell and acellular pertussis vaccines induced an immunity effective at eliminating the genetically different B. pertussis isolates from the lungs. However, the elimination of the PRN-deficient isolate from the lungs of mice vaccinated with commercial vaccines was delayed as compared to the PRN ( +) isolate, suggesting phenotypic differences with the circulating isolates and vaccine strains. The most effective vaccine was the experimental vaccine with the composition identical to that of the strains used for infection.

Corynebacterium diphtheriae infections currently and in the past.

Along with the introduction of common obligatory vaccinations against diphtheria, the disease has been limited in developed countries. However, diphtheria is still endemic in developing countries. Due to a growing popularity of visiting these countries, there is a risk of importation of the disease to Europe. Studies revealed that over 60% of persons aged >40 years in the Polish population do not have a protective level of antibodies against diphtheria. Furthermore, an access to diphtheria antitoxin, which is essential in diphtheria treatment, is now hardly accessible in Europe. On the other hand, in many countries, including Poland, new infections caused by non-toxigenic Corynebacterium diphtheriae have been emerged. Such infections are frequently manifested by bacteraemia and endocarditis with a high fatality rate, amounting even to 41%.

Comparison of Kinetics of Antibody Avidity and IgG Subclasses' Response in Patients with COVID-19 and Healthy Individuals Vaccinated with the BNT162B2 (Comirnaty, Pfizer/BioNTech) mRNA Vaccine.

There are limited reports concerning the levels of antibodies in IgG subclasses and the avidity of IgG, which is the functional strength with which an antibody binds to an antigen in serum samples obtained at different times after infection or vaccination. This study investigated the kinetics of antibody avidity and the IgG antibody response within IgG1-IgG4 subclasses in individuals vaccinated with the BNT162B2 mRNA vaccine and in COVID-19 patients. Serum samples were collected from individuals vaccinated with three doses of the BNT162B2 (Comirnaty, Pfizer/BioNTech) vaccine and from unvaccinated COVID-19 patients. This study revealed that IgG1 was a dominating subclass of IgG both in COVID-19 patients and in vaccinated individuals. The level of IgG4 and IgG avidity significantly increased 7 months after the first two doses of the vaccine and then again after the third dose. IgG2 and IgG3 levels were low in most individuals. Investigating IgG avidity and the dynamics of IgG subclasses is essential for understanding the mechanisms of protection against viral infections, including COVID-19, especially in the context of immunization with innovative mRNA vaccines and the possible future development and application of mRNA technology.

Adjuvant Effect of Whole-Cell Pertussis Component on Tetanus Toxoid Potency in Murine Model.

There is currently an increasing interest in the development of new-generation purified antigen-based vaccines with a higher safety profile compared to conventional inactivated vaccines. The main problem of subunit vaccines is their lower immunogenicity compared to whole-cell vaccines and inducing weaker and shorter-lasting immune responses. In this paper, the results of the assay of the potency of the tetanus component combined with the diphtheria component and whole-cell pertussis vaccine (DTwP), diphtheria and tetanus vaccine (DT), and in monovalent tetanus vaccine (T) are presented. In the mice model, an adjuvant impact of the whole-cell pertussis component on the immune response against tetanus was observed. It was noticed that the potency of tetanus component in the DTwP vaccine was significantly higher than tetanus potency in DT and T vaccines, despite the same bounding ability unit of the tetanus toxoid in the vaccine formulations. The levels of induction of tetanus antibodies by the tested vaccines were also examined. There were no differences in the induction of humoral responses against tetanus by tested vaccines. This publication discusses the possible mechanisms of impact of the whole-cell pertussis component on the other vaccine antigens and the positive and negative aspects of using the whole-cell pertussis component as an adjuvant.

COVID-19 Vaccines over Three Years after the Outbreak of the COVID-19 Epidemic.

The outbreak of COVID-19 started in December 2019 and spread rapidly all over the world. It became clear that the development of an effective vaccine was the only way to stop the pandemic. It was the first time in the history of infectious diseases that the process of the development of a new vaccine was conducted on such a large scale and accelerated so rapidly. At the end of 2020, the first COVID-19 vaccines were approved for marketing. At the end of March 2023, over three years after the outbreak of the COVID-19 pandemic, 199 vaccines were in pre-clinical development and 183 in clinical development. The candidate vaccines in the clinical phase are based on the following platforms: protein subunit, DNA, RNA, non-replication viral vector, replicating viral vector, inactivated virus, virus-like particles, live attenuated virus, replicating viral vector combined with an antigen-presenting cell, non-replication viral vector combined with an antigen-presenting cell, and bacterial antigen-spore expression vector. Some of the new vaccine platforms have been approved for the first time for human application. This review presents COVID-19 vaccines currently available in the world, procedures for assurance of the quality and safety of the vaccines, the vaccinated population, as well as future perspectives for the new vaccine platforms in drug and therapy development for infectious and non-infectious diseases.

Novel combinatorial therapy of oncolytic adenovirus AdV5/3-D24-ICOSL-CD40L with anti PD-1 exhibits enhanced anti-cancer efficacy through promotion of intratumoral T-cell infiltration and modulation of tumour microenvironment in mesothelioma mouse model.

Introduction: Malignant mesothelioma is a rare and aggressive form of cancer. Despite improvements in cancer treatment, there are still no curative treatment modalities for advanced stage of the malignancy. The aim of this study was to evaluate the anti-tumor efficacy of a novel combinatorial therapy combining AdV5/3-D24-ICOSL-CD40L, an oncolytic vector, with an anti-PD-1 monoclonal antibody. Methods: The efficacy of the vector was confirmed in vitro in three mesothelioma cell lines - H226, Mero-82, and MSTO-211H, and subsequently the antineoplastic properties in combination with anti-PD-1 was evaluated in xenograft H226 mesothelioma BALB/c and humanized NSG mouse models. Results and discussion: Anticancer efficacy was attributed to reduced tumour volume and increased infiltration of tumour infiltrating lymphocytes, including activated cytotoxic T-cells (GrB+CD8+). Additionally, a correlation between tumour volume and activated CD8+ tumour infiltrating lymphocytes was observed. These findings were confirmed by transcriptomic analysis carried out on resected human tumour tissue, which also revealed upregulation of CD83 and CRTAM, as well as several chemokines (CXCL3, CXCL9, CXCL11) in the tumour microenvironment. Furthermore, according to observations, the combinatorial therapy had the strongest effect on reducing mesothelin and MUC16 levels. Gene set enrichment analysis suggested that the combinatorial therapy induced changes to the expression of genes belonging to the "adaptive immune response" gene ontology category. Combinatorial therapy with oncolytic adenovirus with checkpoint inhibitors may improve anticancer efficacy and survival by targeted cancer cell destruction and triggering of immunogenic cell death. Obtained results support further assessment of the AdV5/3-D24-ICOSL-CD40L in combination with checkpoint inhibitors as a novel therapeutic perspective for mesothelioma treatment.

Corynebacterium epidermidicanis sp. nov., isolated from skin of a dog.

A Gram-stain-positive, pleomorphic, oxidase-negative, non-motile isolate from the skin of a dog, designated strain 410(T), was subjected to comprehensive taxonomic characterization. Comparison of the 16S rRNA gene sequences revealed that the novel isolate showed highest similarities to the type strains of Corynebacterium humireducens, Corynebacterium diphtheriae, Corynebacterium pseudotuberculosis and Corynebacterium ulcerans (96.1-96.8 %). The quinone system consisted predominantly of MK-8(H(2)) and MK-9(H(2)). The polar lipid profile of strain 410(T) contained the major compounds diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, two unidentified phospholipids and four unidentified glycolipids. The polyamine pattern was composed of the major amines spermidine and spermine. In the fatty acid profile, predominantly straight-chain, saturated and mono-unsaturated fatty acids were detected (C(18 : 1)ω9c, C(16 : 1)ω7c, C(16 : 0)). These chemotaxonomic traits are in agreement with those reported for representatives of the genus Corynebacterium. Strain 410(T) tested negative for diphtheria toxin. Physiological properties as well as unique traits in the polar lipid profile could be used to distinguish strain 410(T) from the most closely related species. These data suggest that strain 410(T) represents a novel species of the genus Corynebacterium, for which we propose the name Corynebacterium epidermidicanis sp. nov. The type strain is 410(T) (= DSM 45586(T) = LMG 26322(T) = CCUG 60915(T)).

[Evaluation of multiplex PCR to identify the species of microorganisms from Bacillus cereus group]. / Ocena przydatnosci metody multiplex PCR do identyfikacji i róznicowania drobnoustrojów z grupy Bacillus cereus.

INTRODUCTION: Bacillus genus comprises about 215 species. The most closely related are B. cereus, B. thuringiensis, B. anthracis, B. mycoides, B. pseudomycoides and B. weihenstephanensis. These bacterial species belong to the Bacillus cereus group. Identification and differentiation of Bacillus cereus group bacteria is difficult because of genetic and phenotypic similarity. Many molecular methods have already been suggested to differentiate B. cereus group members. However easier and more convenient methods are still sought. The aim of this study was to evaluate of multiplex PCR method to identify and distinguish strains of Bacillus cereus group, as proposed by Park et al. (J Microbiol Biotechnol 2007; 17: 1177-82). MATERIALS AND METHOD: Twenty four strains of Bacillus cereus group included B. cereus, B. anthracis, B. thuringiensis and B. weihestephanensis was examined. A multiplex-PCR assay for the differentiation of the species has been applied by using three pairs specific oligonucleotide primers based on sequences of gyrB genes which identify species from Bacillus cereus group and one pair specific primers based on sequences of groEL gene, which are used to identify Bacillus cereus group. RESULTS: Using a specific primers complementary to fragment of groEL gene, we received all PCR products and thus we identified Bacillus cereus group. We have not recived a specific products characteristic for each of the species. Oligonucleotide primers recognized by Park et al. as specific for each species were complementary (often 100%) for the gyrB gene sequence in almost all species of the B. cereus group. CONCLUSIONS: The multiplex PCR method proposed by Park et al. multiplex PCR method for identification ofB. cereus group and individual bacterial species has been proved to be useful only for identification the entire group of B. cereus. This method does not provide specific identification of the individual species. Lack of specificity of the primers used in this study creates a risk of obtaining PCR product in more than one species of the entire examined group of bacteria and does not allow the precise identification to the species level.

[Occurence of pili genes in Corynebacterium diphtheriae strains]. / Wystepowanie genów z wyiazanych z wytwarzaniem pili u szczepów Corynebacterium diphtheriae.

INTRODUCTION: Adhesion of pathogenic bacteria to host cells is a crucial step during infection. The presence of specific adhesion factors on the bacterial cell surface determines the tropism of the pathogen to the tissues expressing certain surface receptors. The adhesion is mediated primarily by filamentous structures called pili. Three distinct pilus structures can be produced by Corynebacterium diphtheriae: SpaA-, SpaD- and SpaH-type pili. Pilus genes encode a total of nine pilus proteins, named SpaA through SpaI, and six sortases, named SrtA through SrtF. All the pilus genes are located on pathogenicity islands and can be acquired and lost by different strains. The aim of presented studies was assessment of occurence of pili genes among C. diphtheriae strains isolated from different infections, including invasive infections, in Poland. METHODS: Thirty-one toxigenic and nontoxigenic C. diphtheriae strains isolated from wounds, blood, nose and pharynx were investigated for presence of 15 pili genes. The studies were conducted using PCR. Gene specific primers were designed on the basis of the complete genome sequence of C. diphtheriae NCTC 13129. RESULT: All the nontoxigenic C. diphtheriae strains isolated from invasive infections possess every tested genes in contrast to toxigenic strains that revealed highly mosaic structure of pili gene clusters. Differences in gene content were detected in SpaA- and SpaH-type pili gene clusters. Complete set of genes in SpaD-type pili gene cluster was detected in all but two strains. The two strains did not possess any of SpaD-type pili genes. CONCLUSIONS: Invasiveness of C. diphtheriae strains could be related to adhesive factors. Results of our studies suggest that ability to express all types of pili is indispensable for causing invasive infections by nontoxigenic C. diphtheriae. Whereas full set ofpili genes is not necessary for causing classical diphtheria.

New Corynebacterium Species with the Potential to Produce Diphtheria Toxin.

Only three Corynebacterium species are known to produce a lethal exotoxin called diphtheria toxin. These are C. diphtheriae, C. ulcerans and C. pseudotuberculosis. The diphtheria toxin gene (tox) is carried in a family of closely related corynebacteriophages and therefore the toxin can be produced only through lysogenisation, in which the corynephage encoding tox is stably inserted into the chromosome. However, 'nontoxigenic tox gene-bearing' (NTTB) strains, which are genotypically tox-positive but do not express the protein, have been described. The emergence of NTTB strains was first observed during the 1990s diphtheria epidemic in Eastern Europe and nowadays such isolates have been detected in many countries in the world. Recently, novel species of Corynebacterium genus have been described which might have the potential of producing the diphtheria toxin due to the possession of the diphtheria toxin gene but it has not produced toxin in laboratory tests. The circulation of NTTB strains could be related to the increased risk for diphtheria disease arising from the risk of re-emerging toxin expression. The article presents the mechanism of diphtheria toxin expression and action, recently described novel species of NTTB corynebacteria as well as the taxonomic changes within the C. diphtheriae group.

Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay.

Rapid and accurate detection and identification of pathogens in clinical samples is essential for all infection diseases. However, in the case of epidemics, it plays a key role not only in the implementation of effective therapy but also in limiting the spread of the epidemic. In this study, we present the application of two nucleic acid isothermal amplification methods-reverse transcription helicase dependent amplification (RT-HDA) and reverse transcription loop-mediated amplification (RT-LAMP)-combined with lateral flow assay as the tools for the rapid detection of SARS-CoV-2, the etiological agent of COVID-19, which caused the ongoing global pandemic. In order to optimize the RT-had, the LOD was 3 genome copies per reaction for amplification conducted for 10-20 min, whereas for RT-LAMP, the LOD was 30-300 genome copies per reaction for a reaction conducted for 40 min. No false-positive results were detected for RT-HDA conducted for 10 to 90 min, but false-positive results occurred when RT-LAMP was conducted for longer than 40 min. We concluded that RT-HDA combined with LFA is more sensitive than RT-LAMP, and it is a good alternative for the development of point-of-care tests for SARS-CoV-2 detection as this method is simple, inexpensive, practical, and does not require qualified personnel to perform the test and interpret its results.

[Microbiological diagnosis of respiratory infections caused by Legionella pneumophila]. / Mikrobiologiczna diagnostyka zakazen ukladu oddechowego wywolywanych przez paleczki legionella pneumophila.

Legionella pneumophila is an important causative agent of pneumonia in humans which is difficult to diagnose because the signs and symptoms are nonspecific and do not distinguish Legionella infection from other common causes of pneumonia. Currently, the diagnosis of Legionnaires' disease is based on phenotyping (culture, antibody detection in human sera, antigen detection in urine) and genotyping methods such as PCR (polymerase chain reaction). This review focuses on current diagnostic tests for surveillance of Legionella pneumophila infections in Poland.

The influence of a swab type on the results of point-of-care tests.

Most point-of-care tests (POCT) use swabs for sampling and/or for applying a sample on the test. A variety of swabs differing in tip materials is commercially available. Different tip materials have different chemical and physical characteristics which might influence the specimen collection and release. We investigated properties of various types of swabs used in clinical diagnostics with focusing on two kinds of analytes, DNA and proteins, which are most often used targets in POCT. As the model samples we used diphtheria toxoid NIBSC 69/017 for investigating recovery of protein analytes such as antigens and bacterial strains of Escherichia coli ATCC 25922, diphtheria toxin-producing Corynebacterium diphtheriae NCTC 10648, and the clinical isolate nontoxigenic C. diphtheriae 5820/15 for investigating the recovery of nucleic acids. We investigated four types of swabs most commonly used in clinical diagnostics in terms of absorption capacity and efficiency of release of nucleic acids and proteins. Volume uptake was measured in milligrams. For DNA release various washing out buffers were used and the amount of released DNA was measured spectrophotometrically. The amount of protein released from the swabs were examined using the Lowry assay. We observed statistically significant differences (p < 0.05) in the mean weights of absorbed liquid, in the DNA recovery and protein recovery by the four variety of swab examined. However, the efficiency of DNA and protein release was not correlated to the absorbed volume of a sample, but rather to the properties of swabs. The swab composition and structure can have a significant impact on the collection and release efficiency of a sample. Therefore, validation of POCT in relation to the used swabs for sampling is really important. The use of inappropriate swabs could lead to false negative or misleading analysis results.

Genetic diversity of Francisella tularensis in Poland with comments on MLVA genotyping and a proposition of a novel rapid v4-genotyping.

We investigated the genotypes of Francisella tularensis (F. tularensis) strains isolated in Poland during the period 1953-2013 and studied their genetic relationship to F. tularensis strains isolated in other countries using MLVA. We examined the mosquito and tick samples collected in Poland for the presence of F. tularensis DNA using PCR. Our results revealed a high genetic diversity among the strains of F. tularensis collected from Poland, suggesting that the bacterium is commonly found in the environment. However, we did not detect F. tularensis DNA in ticks and mosquitoes, showing that the arthropod bites might not be the main source of infection. We also propose the application of a practical assay called v4-genotyping that can be directly performed on the clinical and environmental samples. In addition, we discovered genetic variations among Schu S4 reference strains used in various laboratories and showed that MLVA analysis should not be based on amplicon sizes only because point mutations occurring within the MLVA loci might not always be manifested by a change in the amplicon size.

Analysis of the Amino Acid Sequence Variation of the 67-72p Protein and the Structural Pili Proteins of Corynebacterium diphtheriae for their Suitability as Potential Vaccine Antigens.

The aim of this study was to identify the potential vaccine antigens in Corynebacterium diphtheriae strains by in silico analysis of the amino acid variation in the 67-72p surface protein that is involved in the colonization and induction of epithelial cell apoptosis in the early stages of infection. The analysis of pili structural proteins involved in bacterial adherence to host cells and related to various types of infections was also performed. A polymerase chain reaction (PCR) was carried out to amplify the genes encoding the 67-72p protein and three pili structural proteins (SpaC, SpaI, SapD) and the products obtained were sequenced. The nucleotide sequences of the particular genes were translated into amino acid sequences, which were then matched among all the tested strains using bioinformatics tools. In the last step, the affinity of the tested proteins to major histocompatibility complex (MHC) classes I and II, and linear B-cell epitopes was analyzed. The variations in the nucleotide sequence of the 67-72p protein and pili structural proteins among C. diphtheriae strains isolated from various infections were noted. A transposition of the insertion sequence within the gene encoding the SpaC pili structural proteins was also detected. In addition, the bioinformatics analyses enabled the identification of epitopes for B-cells and T-cells in the conserved regions of the proteins, thus, demonstrating that these proteins could be used as antigens in the potential vaccine development. The results identified the most conserved regions in all tested proteins that are exposed on the surface of C. diphtheriae cells.The aim of this study was to identify the potential vaccine antigens in Corynebacterium diphtheriae strains by in silico analysis of the amino acid variation in the 67­72p surface protein that is involved in the colonization and induction of epithelial cell apoptosis in the early stages of infection. The analysis of pili structural proteins involved in bacterial adherence to host cells and related to various types of infections was also performed. A polymerase chain reaction (PCR) was carried out to amplify the genes encoding the 67­72p protein and three pili structural proteins (SpaC, SpaI, SapD) and the products obtained were sequenced. The nucleotide sequences of the particular genes were translated into amino acid sequences, which were then matched among all the tested strains using bioinformatics tools. In the last step, the affinity of the tested proteins to major histocompatibility complex (MHC) classes I and II, and linear B-cell epitopes was analyzed. The variations in the nucleotide sequence of the 67­72p protein and pili structural proteins among C. diphtheriae strains isolated from various infections were noted. A transposition of the insertion sequence within the gene encoding the SpaC pili structural proteins was also detected. In addition, the bioinformatics analyses enabled the identification of epitopes for B-cells and T-cells in the conserved regions of the proteins, thus, demonstrating that these proteins could be used as antigens in the potential vaccine development. The results identified the most conserved regions in all tested proteins that are exposed on the surface of C. diphtheriae cells.

Pertactin-deficient Bordetella pertussis isolates in Poland-a country with whole-cell pertussis primary vaccination.

The introduction of pertussis vaccination in the 1950s resulted in a significant decrease in the incidence of disease. However, since the 1990s many highly vaccinated countries have observed the re-emergence of the disease. One of the causes of this phenomenon might be related to the adaptation of Bordetella pertussis to vaccination. The purpose of the presented study was an investigation of the emergence and spread of vaccine antigen-deficient B. pertussis isolates in Poland and genomic characterization of the currently circulating pathogen population using PFGE, MLVA and MAST. The results revealed that all tested isolates expressed Ptx, FHA and ACT antigens but 15.4% (4/26) of isolates from 2010 to 2016 were Prn-deficient. Moreover, one TcfA-deficient isolate was collected in 2015. The genotyping showed a genetic distinction between the isolates circulating in 2010-2016 and isolates from previous periods. The majority of currently circulating isolates belonged to PFGE group IV (96.2%), type MT27 (73.1%), and carried ptxA1-ptxC2-ptxP3-prn2-tcfA2-fim2-1-fim3-1 alleles (61.5%). The unique genetic structure of the B. pertussis population in Poland has changed since 2010 and became similar to that observed in countries with aP vaccination. This could be a result of increasing use of aP vaccines (60% of primary vaccination in 2013) over wP vaccines, which have been broadly used for primary vaccination in Poland for decades.

[Types of Corynebacterium diphtheriae strains isolated in Poland in 2004-2008]. / Typy szczepów Corynebacterium diphtheriae izolowanych w Polsce w latach 2004-2008.

Biotyping and genotyping were performed for Corynebacterium diphtheriae strains isolated from humans in Poland during last several years. All the strains belonged to the same biotype and genetic clone. Similar situation has not yet been described in international literature, although nontoxigenic C. diphtheriae strains have been isolated more frequently in developed countries. However, strains isolated and investigated in other countries revealed heterogeneity of biotypes and genotypes.

[Valuation for usefulness of selected chromosomal markers for Bacillus anthracis identification. I. Valuation for markers SG-749, SG-450 and SG-300]. / Ocena przydatnosci wybranych chromosomalnych markerów do identyfikacji Bacillus anthracis. I. Ocena markerów SG-749, SG-450 i SG-300.

The article presents results of valuation for B. anthracis-specificity and usefulness for its identification obtained for different chromosomal markers. In the first part of the study markers SG-749, SG-300 and SG-450 were analyzed. For the investigation RFLP-PCR and MSSCP techniques were used and different electrophoresis methods were tested. The results gave an information not only about specificity of tested markers but also about the possibility of shorten time necessary to obtain results of B. anthracis identification.

[Valuation for usefulness of selected chromosomal markers for Bacillus anthracis identification. II. Valuation for markers SSH and rpoB]. / Ocena przydatnosci wybranych chromosomalnych markerów do identyfikacji Bacillus anthracis. II. Ocena markerów SSH oraz rpoB.

The article presents results of valuation for B. anthracis-specificity and usefulness for its identification obtained for different chromosomal markers. In the second part of the study markers SSH241, SSH196, SSH163, SSH133 as well as a fragment of the house-keeping gene rpoB were analyzed. For the investigation MSSCP and multiplex-PCR assays were used. There were also tested different techniques of electrophoresis. The results gave an information about specificity of tested markers and their usefulness for B. anthracis identification.