The aim of the study was to propose a multilocus calibrator containing seven specific sequences of SSTR1-SSTR5, D2R, ER1 genes and the GUS house-keeping reference gene. For preparation, gene synthesis and gene assembly technique were used. The experimental group consisted of 44 pituitary adenomas with overproduction of growth hormone leading to acromegaly. The gene expression was investigated using the real-time PCR. We revealed highest numbers of SSTR2 and D2R transcripts, lower numbers of SSTR1, SSTR3, SSTR5 and ER1 trans¬cripts, and the lowest transcription of the SSTR4 gene. The use of the multicalibrator ensured a high comparability of the results obtained for the tested genes.

• MeSH
• akromegalie * etiologie   genetika   MeSH
• biopsie MeSH
• dospělí MeSH
• exprese genu MeSH
• genetická transkripce MeSH
• insulinu podobný růstový faktor I sekrece   MeSH
• kalibrace * MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• multilokusová sekvenční typizace MeSH
• nádory hypofýzy * genetika   MeSH
• receptory dopaminové * analýza   MeSH
• receptory somatostatinu * analýza   MeSH
• růstový hormon sekrece   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• klinická studie MeSH
• práce podpořená grantem MeSH

To date, genotyping data on giardiasis have not been available in the Czech Republic. In this study, we characterized 47 human isolates of Giardia intestinalis from symptomatic as well as asymptomatic giardiasis cases. Genomic DNA from trophozoites was tested by PCR-sequence analysis at three loci (β-giardin, glutamate dehydrogenase and triose phosphate isomerase). Sequence analysis showed assemblages A and B in 41 (87.2%) and six (12.8%) isolates, respectively. Two of the 41 assemblage A samples were genotyped as sub-assemblage AI, and 39 were genotyped as sub-assemblage AII. Four previously identified multilocus genotypes (MLGs: AI-1, AII-1, AII-4 and AII-9) and six likely novel variations of MLGs were found. In agreement with previous studies, sequences from assemblage B isolates were characterized by a large genetic variability and by the presence of heterogeneous positions, which prevent the definition of MLGs. This study also investigated whether there was a relationship between the assemblage and clinical data (including drug resistance). However, due to the large number of genotypes and the relatively small number of samples, no significant associations with the clinical data were found.

• MeSH
• dítě MeSH
• dospělí MeSH
• genotyp MeSH
• Giardia klasifikace   enzymologie   genetika   MeSH
• giardiáza parazitologie   MeSH
• glutamátdehydrogenasa genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• multilokusová sekvenční typizace MeSH
• předškolní dítě MeSH
• protozoální proteiny genetika   MeSH
• sekvenční analýza DNA MeSH
• triózafosfátizomeráza genetika   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Treponema pallidum strains are closely related at the genome level but cause distinct diseases. Subspecies pallidum (TPA) is the causative agent of syphilis, subspecies pertenue (TPE) causes yaws while subspecies endemicum (TEN) causes bejel (endemic syphilis). Compared to the majority of treponemal genomic regions, several chromosomal loci were found to be more diverse. To assess genetic variability in diverse genomic positions, we have selected (based on published genomic data) and sequenced five variable loci, TP0304, TP0346, TP0488, TP0515 and TP0558, in 19 reference Treponema pallidum strains including all T. pallidum subspecies (TPA, TPE and TEN). Results of this multilocus analysis divided syphilitic isolates into two groups: SS14-like and Nichols-like. The SS14-like group is comprised of SS14, Grady, Mexico A and Philadelphia 1 strains. The Nichols-like group consisted of strains Nichols, Bal 73-1, DAL-1, MN-3, Philadelphia 2, Haiti B and Madras. The TP0558 locus was selected for further studies because it clearly distinguished between the SS14- and Nichols-like groups and because the phylogenetic tree derived from the TP0558 locus showed the same clustering pattern as the tree constructed from whole genome sequences. In addition, TP0558 was shown as the only tested locus that evolved under negative selection within TPA strains. Sequencing of a short fragment (573bp) of the TP0558 locus in a set of 25 clinical isolates from 22 patients collected in the Czech Republic during 2012-2013 revealed that clinical isolates follow the SS14- and Nichols-like distribution.

• MeSH
• dospělí MeSH
• genotyp MeSH
• lidé středního věku MeSH
• lidé MeSH
• molekulární epidemiologie MeSH
• multilokusová sekvenční typizace MeSH
• novorozenec MeSH
• shluková analýza MeSH
• syfilis epidemiologie   mikrobiologie   MeSH
• Treponema pallidum klasifikace   genetika   izolace a purifikace   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika 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

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.

• 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

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 MeSH
• Švýcarsko MeSH

"Candidatus Neoehrlichia mikurensis" is the tick-borne agent of neoehrlichiosis, an infectious disease that primarily affects immunocompromised patients. So far, the genetic variability of "Ca. Neoehrlichia" has been studied only by comparing 16S rRNA genes and groEL operon sequences. We describe the development and use of a multilocus sequence analysis (MLSA) protocol to characterize the genetic diversity of clinical "Ca. Neoehrlichia" strains in Europe and their relatedness to other species within the Anaplasmataceae family. Six genes were selected: ftsZ, clpB, gatB, lipA, groEL, and 16S rRNA. Each MLSA locus was amplified by real-time PCR, and the PCR products were sequenced. Phylogenetic trees of MLSA locus relatedness were constructed from aligned sequences. Blood samples from 12 patients with confirmed "Ca. Neoehrlichia" infection from Sweden (n = 9), the Czech Republic (n = 2), and Germany (n = 1) were analyzed with the MLSA protocol. Three of the Swedish strains exhibited identical lipA sequences, while the lipA sequences of the strains from the other nine patients were identical to each other. One of the Czech strains had one differing nucleotide in the clpB sequence from the sequences of the other 11 strains. All 12 strains had identical sequences for the genes 16S rRNA, ftsZ, gatB, and groEL. According to the MLSA, among the Anaplasmataceae, "Ca. Neoehrlichia" is most closely related to Ehrlichia ruminantium, less so to Anaplasma phagocytophilum, and least to Wolbachia endosymbionts. To conclude, three sequence types of infectious "Ca. Neoehrlichia" were identified: one in the west of Sweden, one in the Czech Republic, and one spread throughout Europe.

• MeSH
• Anaplasmataceae klasifikace   genetika   izolace a purifikace   MeSH
• esenciální geny MeSH
• fylogeneze MeSH
• genetická variace * MeSH
• genotyp * MeSH
• infekce bakteriemi čeledi Anaplasmataceae epidemiologie   mikrobiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• molekulární epidemiologie metody   MeSH
• multilokusová sekvenční typizace metody   MeSH
• RNA ribozomální 16S genetika   MeSH
• senioři MeSH
• shluková analýza MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Německo MeSH
• Švédsko MeSH

Pro klonální analýzu kmenů Streptococcus pneumoniae z invazivního pneumokokového onemocnění byla v NRLpro streptokokové nákazy SZÚ zavedena a ověřena metoda Multiple-Locus Variable number tandem repeat Ana-lysis (MLVA). Oproti rutinně používané metodě Multilocus Sequence Typing (MLST) je výhodou metody MLVAsnadnost a rychlost provedení i nižší nákladnost. Rozlišení klonů je u obou metod obdobné. Standardem klonálníanalýzy pro molekulární surveillance původců závažných pneumokokových infekcí je však MLST, MLVA můžebýt doplňkem pro klonální porovnání izolátů z lokálních epidemiologických situací.

The NRL for Streptococcal Infections implemented and tested Multiple-Locus Variable number tandem repeatAnalysis (MLVA) for clonal analysis of Streptococcus pneumoniae strains from invasive pneumococcal disease.As compared to the routinely used Multilocus Sequence Typing (MLST) method, MLVA has the advantage of beingeasier, faster, and less expensive to perform. Both methods have similar resolution. Nevertheless, the standardmethod for clonal analysis in molecular surveillance of causative agents of severe pneumococcal infections isMLST, and MLVA can be used as a complementary method for clonal comparison of local isolates.

• MeSH
• DNA bakterií genetika   MeSH
• minisatelitní repetice genetika   MeSH
• multilokusová sekvenční typizace * metody   MeSH
• pneumokokové infekce diagnóza   epidemiologie   MeSH
• Streptococcus pneumoniae * genetika   izolace a purifikace   MeSH
• techniky typizace bakterií metody   MeSH
• Publikační typ
• práce podpořená grantem MeSH

The taxonomy and systematics of the armored harvestmen (suborder Laniatores) are based on various sets of morphological characters pertaining to shape, armature, pedipalpal setation, and the number of articles of the walking leg tarsi. Few studies have tested the validity of these historical character systems in a comprehensive way, with reference to an independent data class, i.e., molecular sequence data. We examined as a test case the systematics of Podoctidae, a family distributed throughout the Indo-Pacific. We tested the validity of the three subfamilies of Podoctidae using a five-locus phylogeny, and examined the evolution of dorsal shape as a proxy for taxonomic utility, using parametric shape analysis. Here we show that two of the three subfamilies, Ibaloniinae and Podoctinae, are non-monophyletic, with the third subfamily, Erecananinae, recovered as non-monophyletic in a subset of analyses. Various genera were also recovered as non-monophyletic. As first steps toward revision of Podoctidae, the subfamilies Erecananinae Roewer, 1912 and Ibaloniinae Roewer, 1912 are synonymized with Podoctinae Roewer, 1912 new synonymies, thereby abolishing unsubstantiated subfamilial divisions within Podoctidae. We once again synonymize the genus Paralomanius Goodnight & Goodnight, 1948 with Lomanius Roewer, 1923 revalidated. We additionally show that eggs carried on the legs of male Podoctidae are not conspecific to the males, falsifying the hypothesis of paternal care in this group.

• MeSH
• Bayesova věta MeSH
• cytochromy c klasifikace   genetika   metabolismus   MeSH
• DNA chemie   izolace a purifikace   metabolismus   MeSH
• fylogeneze MeSH
• ovum metabolismus   MeSH
• pavoukovci anatomie a histologie   klasifikace   genetika   růst a vývoj   MeSH
• RNA ribozomální 16S klasifikace   genetika   metabolismus   MeSH
• sekvenční analýza DNA MeSH
• sekvenční seřazení MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. 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