BACKGROUND: The increased prevalence of syphilis in Cuba prompted us to map the circulating Treponema pallidum subsp. pallidum allelic profiles in this geographic region. METHODS: Samples were collected from 2012 to 2017, from 83 male patients with ulcers or skin lesions, and were examined using multilocus sequence typing. Additionally, we analyzed the 23S rDNA and 16S rDNA regions for the presence of possible mutations leading to macrolide and tetracycline resistance. RESULTS: Among 94% of fully typed strains, we found 7 different allelic profiles, of which 4 had not been previously described. More than 87% of patients were infected with the T. pallidum SS14-like group and only 8.2% with T. pallidum Nichols-like group. As in other countries, the 1.3.1 allelic profile (ie, SS14-like) was the most common. In addition, 1 of the newly described allelic profiles represents T. pallidum strains that arose by recombination events between members of different T. pallidum subgroups. More than 90% of patients were infected with treponemes harboring the A2058G mutation. However, we found no potential tetracycline-resistant T. pallidum mutations. CONCLUSIONS: Our results suggest that, in Cuba, tetracycline antibiotics could be used to treat syphilis in penicillin-allergic patients instead of macrolides.

• Klíčová slova
• Treponema pallidum subsp. pallidum, Cuba, macrolide resistance, multilocus sequence typing, recombination in treponemes,
• MeSH
• alely MeSH
• antibakteriální látky MeSH
• bakteriální léková rezistence genetika   MeSH
• dospělí MeSH
• lidé MeSH
• makrolidy terapeutické užití   MeSH
• multilokusová sekvenční typizace MeSH
• mutace MeSH
• syfilis farmakoterapie   mikrobiologie   MeSH
• techniky typizace bakterií MeSH
• tetracyklin terapeutické užití   MeSH
• Treponema pallidum klasifikace   genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Kuba MeSH
• Názvy látek
• antibakteriální látky MeSH
• makrolidy MeSH
• tetracyklin MeSH

The MLST scheme currently used for Enterococcus faecium typing was designed in 2002 and is based on putative gene functions and Enterococcus faecalis gene sequences available at that time. As a result, the original MLST scheme does not correspond to the real genetic relatedness of E. faecium strains and often clusters genetically distant strains to the same sequence types (ST). Nevertheless, typing has a significant impact on the subsequent epidemiological conclusions and introduction of appropriate epidemiological measures, thus it is crucial to use a more accurate MLST scheme. Based on the genome analysis of 1,843 E. faecium isolates, a new scheme, consisting of 8 highly discriminative loci, was created in this study. These strains were divided into 421 STs using the new MLST scheme, as opposed to 223 STs assigned by the original MLST scheme. The proposed MLST has a discriminatory power of D = 0.983 (CI95% 0.981 to 0.984), compared to the original scheme's D = 0.919 (CI95% 0.911 to 0.927). Moreover, we identified new clonal complexes with our newly designed MLST scheme. The scheme proposed here is available within the PubMLST database. Although whole genome sequencing availability has increased rapidly, MLST remains an integral part of clinical epidemiology, mainly due to its high standardization and excellent robustness. In this study, we proposed and validated a new MLST scheme for E. faecium, which is based on genome-wide data and thus reflects the tested isolates' more accurate genetic similarity. IMPORTANCE Enterococcus faecium is one of the most important pathogens causing health care associated infections. One of the main reasons for its clinical importance is a rapidly spreading resistance to vancomycin and linezolid, which significantly complicates antibiotic treatment of infections caused by such resistant strains. Monitoring the spread and relationships between resistant strains causing severe conditions represents an important tool for implementing appropriate preventive measures. Therefore, there is an urgent need to establish a robust method enabling strain monitoring and comparison at the local, national, and global level. Unfortunately, the current, extensively used MLST scheme does not reflect the real genetic relatedness between individual strains and thus does not provide sufficient discriminatory power. This can lead directly to incorrect epidemiological measures due to insufficient accuracy and biased results.

• Klíčová slova
• Enterococcus faesium, clonal complex, epidemiology, multilocus sequence typing, whole genome sequenging,
• MeSH
• antibakteriální látky MeSH
• Enterococcus faecium * genetika   MeSH
• grampozitivní bakteriální infekce * epidemiologie   MeSH
• lidé MeSH
• multilokusová sekvenční typizace metody   MeSH
• sekvenování celého genomu MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antibakteriální látky MeSH

Cystic fibrosis (CF) patients in the Czech Republic suffered in the late 1990s from an epidemic with ST32 strain of Burkholderia cepacia complex (Bcc). Cohort segregation of Bcc and of ST32 positive patients was introduced in 1999 and 2002, respectively. We performed a study to evaluate the molecular epidemiology of Bcc infection after implementation of these infection control measures. Patients attending the Prague CF adult Centre from 2000 to 2015 were included in the present study. Demographic data and microbial statuses were collected from patient records. All Bcc isolates were analyzed using multilocus sequence typing (MLST). The prevalences of epidemic strain ST32 and of other Bcc strains were calculated. Ninety out of 227 CF patients were infected with Bcc during the study period. The prevalence of ST32 cases significantly decreased from 46.5% in 2000-2001 to 10.4% in 2014-2015 (P < 0.001) due to occurrence of only one new case in 2003, as well as to the death of 72% of ST32-infected patients. Conversely, there was a significant increase in prevalence of other Bcc strains, which rose from 0 to 14.9% (P = 0.015) and of transient infections. A micro-epidemic of infection with ST630 strain was observed in 2014 in lung transplant patients hospitalized in intensive care unit. The prevalence of epidemic strain ST32 decreased, whereas that of non-clonal strains of Bcc increased. Routine use of MLST allowed early detection of new and potentially epidemic strains.

• Klíčová slova
• Cystic Fibrosis, Cystic Fibrosis Patient, Molecular Genetic Method, Multilocus Sequence Typing, recA Group,
• MeSH
• Burkholderia cepacia komplex klasifikace   genetika   izolace a purifikace   MeSH
• cystická fibróza epidemiologie   mikrobiologie   MeSH
• infekce bakteriemi rodu Burkholderia epidemiologie   mikrobiologie   MeSH
• lidé MeSH
• molekulární epidemiologie MeSH
• multilokusová sekvenční typizace MeSH
• prevalence MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

Cronobacter spp. are bacterial pathogens that affect children and immunocompromised adults. In this study, we used multilocus sequence typing (MLST) to determine sequence types (STs) in 11 Cronobacter spp. strains isolated from retail foods, 29 strains from dust samples obtained from vacuum cleaners, and 4 clinical isolates. Using biochemical tests, species-specific polymerase chain reaction, and MLST analysis, 36 strains were identified as Cronobacter sakazakii, and 6 were identified as Cronobacter malonaticus. In addition, one strain that originated from retail food and one from a dust sample from a vacuum cleaner were identified on the basis of MLST analysis as Cronobacter dublinensis and Cronobacter turicensis, respectively. Cronobacter spp. strains isolated from the retail foods were assigned to eight different MLST sequence types, seven of which were newly identified. The strains isolated from the dust samples were assigned to 7 known STs and 14 unknown STs. Three clinical isolates and one household dust isolate were assigned to ST4, which is the predominant ST associated with neonatal meningitis. One clinical isolate was classified based on MLST analysis as Cronobacter malonaticus and belonged to an as-yet-unknown ST. Three strains isolated from the household dust samples were assigned to ST1, which is another clinically significant ST. It can be concluded that Cronobacter spp. strains of different origin are genetically quite variable. The recovery of C. sakazakii strains belonging to ST1 and ST4 from the dust samples suggests the possibility that contamination could occur during food preparation. All of the novel STs and alleles for C. sakazakii, C. malonaticus, C. dublinensis, and C. turicensis determined in this study were deposited in the Cronobacter MLST database available online ( http://pubmlst.org/cronobacter/).

• MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• Cronobacter sakazakii klasifikace   izolace a purifikace   metabolismus   MeSH
• Cronobacter klasifikace   izolace a purifikace   metabolismus   MeSH
• enterobakteriální infekce mikrobiologie   MeSH
• fylogeneze MeSH
• lidé MeSH
• mikrobiologie životního prostředí * MeSH
• molekulární typizace MeSH
• multilokusová sekvenční typizace MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• potravinářská mikrobiologie * MeSH
• prach * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• bakteriální proteiny MeSH
• prach * MeSH

The first multilocus sequence typing results of Czech neonatal Streptococcus agalactiae isolates are presented in this paper. The aims of the study were to prove if the Czech isolates belong to potential invasive clonal complex CC17 and to further analyse serotype-sequence type relationships. Twelve sequence types detected among 77 isolates were assigned to 5 clonal complexes. Sequence type variability was found for most of our collection serotypes. As many as 83 % of the invasive isolates were covered by as few as 3 sequence types: S17, ST23 and ST19. ST17 was the only sequence type strictly tied to serotype III. "Hyperinvasive" ST17 was identified in 85 % of the cerebrospinal fluid isolates (6 out of 7), but in 32 % of the blood isolates only.

• MeSH
• lidé MeSH
• multilokusová sekvenční typizace * MeSH
• novorozenec MeSH
• Streptococcus agalactiae genetika   izolace a purifikace   MeSH
• streptokokové infekce mikrobiologie   MeSH
• techniky typizace bakterií * MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• centra pro kontrolu nemocí a prevenci (USA) MeSH
• lidé MeSH
• multilokusová sekvenční typizace MeSH
• syfilis * MeSH
• Treponema pallidum genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• komentáře MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Spojené státy americké MeSH

Syphilis is an important public health problem and an increasing incidence has been noted in recent years. Characterization of strain diversity through molecular data plays a critical role in the epidemiological understanding of this re-emergence. We here propose a new high-resolution multilocus sequence typing (MLST) scheme for Treponema pallidum subsp. pallidum (TPA). We analyzed 30 complete and draft TPA genomes obtained directly from clinical samples or from rabbit propagated strains to identify suitable typing loci and tested the new scheme on 120 clinical samples collected in Switzerland and France. Our analyses yielded three loci with high discriminatory power: TP0136, TP0548, and TP0705. Together with analysis of the 23S rRNA gene mutations for macrolide resistance, we propose these loci as MLST for TPA. Among clinical samples, 23 allelic profiles as well as a high percentage (80% samples) of macrolide resistance were revealed. The new MLST has higher discriminatory power compared to previous typing schemes, enabling distinction of TPA from other treponemal bacteria, distinction between the two main TPA clades (Nichols and SS14), and differentiation of strains within these clades.

• MeSH
• alely MeSH
• antibakteriální látky farmakologie   MeSH
• DNA bakterií genetika   MeSH
• fylogeneze MeSH
• genom bakteriální MeSH
• genotyp MeSH
• globus pallidus MeSH
• jednonukleotidový polymorfismus MeSH
• makrolidy farmakologie   MeSH
• multilokusová sekvenční typizace metody   MeSH
• RNA ribozomální 23S genetika   MeSH
• sekvenční analýza DNA metody   MeSH
• syfilis epidemiologie   MeSH
• Treponema pallidum genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Francie epidemiologie   MeSH
• Švýcarsko epidemiologie   MeSH
• Názvy látek
• antibakteriální látky MeSH
• DNA bakterií MeSH
• makrolidy MeSH
• RNA ribozomální 23S MeSH

BACKGROUND: Out of 20 spirochete species from Borrelia burgdorferi sensu lato (s.l.) complex recognized to date some are considered to have a limited distribution, while others are worldwide dispersed. Among those are Borrelia burgdorferi sensu stricto (s.s.) and Borrelia bissettii which are distributed both in North America and in Europe. While B. burgdorferi s.s. is recognized as a cause of Lyme borreliosis worldwide, involvement of B. bissettii in human Lyme disease was not so definite yet. FINDINGS: Multilocus sequence typing of spirochete isolates originating from residents of Georgia and Florida, USA, revealed the presence of two Borrelia burgdorferi sensu stricto strains highly similar to those from endemic Lyme borreliosis regions of the northeastern United States, and an unusual strain that differed from any previously described in Europe or North America. Based on phylogenetic analysis of eight chromosomally located housekeeping genes divergent strain clustered between Borrelia bissettii and Borrelia carolinensis, two species from the B.burgdorferi s.l. complex, widely distributed among the multiple hosts and vector ticks in the southeastern United States. The genetic distance analysis showed a close relationship of the diverged strain to B. bissettii. CONCLUSIONS: Here, we present the analysis of the first North American human originated live spirochete strain that revealed close relatedness to B. bissettii. The potential of B. bissettii to cause human disease, even if it is infrequent, is of importance for clinicians due to the extensive range of its geographic distribution.

• MeSH
• Borrelia burgdorferi komplex MeSH
• Borrelia burgdorferi MeSH
• Borrelia klasifikace   genetika   izolace a purifikace   MeSH
• esenciální geny MeSH
• genotyp MeSH
• lidé MeSH
• lymeská nemoc diagnóza   mikrobiologie   MeSH
• molekulární sekvence - údaje MeSH
• multilokusová sekvenční typizace * MeSH
• sekvenční analýza DNA MeSH
• shluková analýza MeSH
• Spirochaetales MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Florida MeSH
• Georgia MeSH

MLST typing of Neisseria meningitidis directly from clinical material was introduced in the National Reference Laboratory for Meningococcal Infections in Prague. Four cerebrospinal fluid samples were obtained from patients with suspected meningococcal invasive disease. In all samples, all classical laboratory methods gave negative results and the only positive method was PCR, which revealed Neisseria meningitidis group C (two specimens) and group B (two specimens), respectively. MLST performed directly from cerebrospinal fluid revealed that the strains causing the two group C infections were of sequence type (ST) 11, while the two group B infections were characterized as ST-32 and ST-33 respectively. Multi-locus sequence typing of meningococci directly from clinical material offers the opportunity to improve further the surveillance of meningococcal disease and has now been introduced into the routine portfolio of tests employed at the national reference laboratory of the Czech Republic.

• MeSH
• DNA bakterií MeSH
• DNA primery MeSH
• lidé MeSH
• meningokokové infekce patologie   MeSH
• mladiství MeSH
• mozkomíšní mok mikrobiologie   MeSH
• Neisseria meningitidis genetika   izolace a purifikace   patogenita   MeSH
• polymerázová řetězová reakce MeSH
• sekvenční analýza DNA MeSH
• surveillance populace MeSH
• Check Tag
• lidé MeSH
• mladiství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA bakterií MeSH
• DNA primery MeSH

Given the great biological importance and high diversity of temperate Staphylococcus aureus bacteriophages, a method is needed for the description of their genomic structure. Here we have updated a multiplex PCR strategy for the complex characterization of S. aureus phages of the family Siphoviridae. Based on the comparative genomic analysis of the available phage sequences, a multilocus PCR strategy for typing the major modules of the phage genome was designed. The genomic modules were classified on the basis of the genes for integrase (10 types), anti-repressor (five types), replication proteins polA, dnaC and dnaD (four types), dUTPase (four types), portal protein (eight types), tail appendices (four types) and endolysin (four types) corresponding to the integrase locus, lysogeny control region, and modules for DNA replication, transcription regulation, packaging, tail appendices and lysis respectively. The nine PCR assays designed for the above sequences were shown to be capable to identify the bacteriophage gene pool present both in the phage and bacterial genomes and their extensive mosaic structure. The established multiplex PCR-based multilocus diagnostic scheme is convenient for rapid and reliable phage and prophage classification and for the study of bacteriophage evolution.

• MeSH
• DNA virů genetika   MeSH
• genom virový * MeSH
• multilokusová sekvenční typizace MeSH
• polymerázová řetězová reakce metody   MeSH
• profágy klasifikace   genetika   MeSH
• Siphoviridae klasifikace   genetika   MeSH
• srovnávací genomová hybridizace MeSH
• Staphylococcus aureus genetika   virologie   MeSH
• virové proteiny genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA virů MeSH
• virové proteiny MeSH