Determination of the causative agent of erythema-like skin lesions in case of nonspecific superficial perivascular dermatitis was supported by histological examination and led to the latter diagnosis of Hyperkeratosis lenticularis perstans (Flegel disease) in patient. The presence of antibodies against Borrelia burgdorferi in patient serum was confirmed by a routine ELISA method and verified by Western blot technique. Skin biopsy and blood specimens were analyzed by PCR and multilocus sequence analysis (MLSA). Western blot method revealed IgG antibody response against two specific antigens, 17 and 83 kDa proteins. The recombinant test detected IgG antibody response against p100 and p41 antigens. The sequence analysis of amplicons from the selected genomic loci obtained from skin biopsy and serum samples revealed the presence of two species from B. burgdorferi sensu lato complex as a co-infection in this patient-B. burgdorferi sensu stricto (s.s.) and Borrelia garinii.

• MeSH
• antigeny bakteriální imunologie   MeSH
• biopsie MeSH
• Borrelia burgdorferi komplex genetika   izolace a purifikace   MeSH
• Borrelia burgdorferi genetika   izolace a purifikace   MeSH
• DNA bakterií genetika   izolace a purifikace   MeSH
• imunoglobulin G krev   MeSH
• keratóza mikrobiologie   patologie   MeSH
• koinfekce mikrobiologie   patologie   MeSH
• kůže mikrobiologie   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• multilokusová sekvenční typizace MeSH
• polymerázová řetězová reakce MeSH
• protilátky bakteriální krev   MeSH
• western blotting MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• Názvy látek
• antigeny bakteriální MeSH
• DNA bakterií MeSH
• imunoglobulin G MeSH
• protilátky bakteriální MeSH

BACKGROUND: The controversy surrounding the potential impact of birds in spirochete transmission dynamics and their capacity to serve as a reservoir has existed for a long time. The majority of analyzed bird species are able to infect larval ticks with Borrelia. Dispersal of infected ticks due to bird migration is a key to the establishment of new foci of Lyme borreliosis. The dynamics of infection in birds supports the mixing of different species, the horizontal exchange of genetic information, and appearance of recombinant genotypes. METHODS: Four Borrelia burgdorferi sensu lato strains were cultured from Ixodes minor larvae and four strains were isolated from Ixodes minor nymphs collected from a single Carolina Wren (Thryothorus ludovicianus). A multilocus sequence analysis that included 16S rRNA, a 5S-23S intergenic spacer region, a 16S-23S internal transcribed spacer, flagellin, p66, and ospC separated 8 strains into 3 distinct groups. Additional multilocus sequence typing of 8 housekeeping genes, clpA, clpX, nifS, pepX, pyrG, recG, rplB, and uvrA was used to resolve the taxonomic status of bird-associated strains. RESULTS: Results of analysis of 14 genes confirmed that the level of divergence among strains is significantly higher than what would be expected for strains within a single species. The presence of cross-species recombination was revealed: Borrelia burgdorferi sensu stricto housekeeping gene nifS was incorporated into homologous locus of strain, previously assigned to B. americana. CONCLUSIONS: Genetically diverse Borrelia strains are often found within the same tick or same vertebrate host, presenting a wide opportunity for genetic exchange. We report the cross-species recombination that led to incorporation of a housekeeping gene from the B. burgdorferi sensu stricto strain into a homologous locus of another bird-associated strain. Our results support the hypothesis that recombination maintains a majority of sequence polymorphism within Borrelia populations because of the re-assortment of pre-existing sequence variants. Even if our findings of broad genetic diversity among 8 strains cultured from ticks that fed on a single bird could be the exception rather than the rule, they support the theory that the diversity and evolution of LB spirochetes is driven mainly by the host.

• MeSH
• Borrelia burgdorferi klasifikace   genetika   izolace a purifikace   MeSH
• esenciální geny MeSH
• fylogeneze MeSH
• genetická variace MeSH
• klíšťata mikrobiologie   MeSH
• molekulární sekvence - údaje MeSH
• ptáci mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Borrelia burgdorferi sensu lato (s.l.) complex is a diverse group of worldwide distributed bacteria that includes 18 named spirochete species and a still not named group proposed as genomospecies 2. Descriptions of new species and variants continue to be recognized, so the current number of described species is probably not final. Most of known spirochete species are considered to have a limited distribution. Eleven species from the B. burgdorferi s.l. complex were identified in and strictly associated with Eurasia (B. afzelii, B. bavariensis, B. garinii, B. japonica, B. lusitaniae, B. sinica, B. spielmanii, B. tanukii, B. turdi, B. valaisiana, and B. yangtze), while another 5 (B. americana, B. andersonii, B. californiensis, B. carolinensis, and B. kurtenbachii) were previously believed to be restricted to the USA only. B. burgdorferi sensu stricto (s.s.), B. bissettii, and B. carolinensis share the distinction of being present in both the Old and the New World. Out of the 18 genospecies, 3 commonly and 4 occasionally infect humans, causing Lyme borreliosis (LB) - a multisystem disease that is often referred to as the 'great imitator' due to diversity of its clinical manifestations. Among the genospecies that commonly infect people, i.e. B. burgdorferi s.s., B. afzelii, and B. garinii, only B. burgdorferi s.s. causes LB both in the USA and in Europe, with a wide spectrum of clinical conditions ranging from minor cutaneous erythema migrans (EM) to severe arthritis or neurological manifestations. The epidemiological data from many European countries and the USA show a dramatic increase of the diagnosed cases of LB due to the development of new progressive diagnostic methods during the last decades (Hubálek, 2009). Recently, the definition of the disease has also changed. What was not considered Lyme borreliosis before might be now.

• MeSH
• arachnida jako vektory mikrobiologie   fyziologie   MeSH
• Borrelia burgdorferi klasifikace   genetika   izolace a purifikace   patogenita   MeSH
• DNA bakterií genetika   MeSH
• druhová specificita MeSH
• fylogeneze MeSH
• fylogeografie MeSH
• genetická variace MeSH
• hlodavci MeSH
• klíšťata mikrobiologie   fyziologie   MeSH
• lidé MeSH
• lymeská nemoc * diagnóza   epidemiologie   mikrobiologie   přenos   MeSH
• polymerázová řetězová reakce MeSH
• ptáci MeSH
• techniky typizace bakterií metody   MeSH
• veřejné zdravotnictví MeSH
• zdroje nemoci MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Geografické názvy
• Asie MeSH
• Evropa MeSH
• Spojené státy americké MeSH
• Názvy látek
• DNA bakterií MeSH

A multiplex PCR assay was developed for the detection of toxigenic and pathogenic V. cholerae from direct water sources using specific primers targeting diverse genes, viz. outer membrane protein (ompW), cholera toxin (ctxB), ORF specific for O1 (rfbG), zonula occludens (zot) and toxin co-regulated pilus (tcpB); among these genes, ompW acts as internal control for V. cholerae, the ctx gene as a marker for toxigenicity and tcp for pathogenicity. The sensitivity of multiplex PCR was 5 x 10(4) V. cholerae cells per reaction. The procedure was simplified as direct bacterial cells were used as template and there was no need for DNA extraction. The assay was specific as no amplification occurred with the other bacteria used. Toxigenic V. cholerae were artificially spiked in different water samples, filtered through a 0.45 microm membrane, and the filters containing bacteria were enriched in APW for 6 h. PCR following filtration and enrichment could detect as little as 8 V. cholerae cells per mL in different spiked water samples. Various environmental potable water samples were screened for the presence of V. cholerae using this assay procedure. The proposed method is rapid, sensitive and specific for environmental surveillance for the presence of toxigenic-pathogenic and nonpathogenic V. cholerae.

• MeSH
• bakteriální proteiny genetika   MeSH
• cholerový toxin genetika   MeSH
• DNA primery MeSH
• látky znečišťující vodu analýza   MeSH
• monitorování životního prostředí metody   MeSH
• polymerázová řetězová reakce metody   MeSH
• senzitivita a specificita MeSH
• sladká voda mikrobiologie   MeSH
• Vibrio cholerae non-O1 * genetika   izolace a purifikace   patogenita   MeSH
• Vibrio cholerae O1 * genetika   izolace a purifikace   patogenita   MeSH
• zásobování vodou analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• bakteriální proteiny MeSH
• cholerový toxin MeSH
• DNA primery MeSH
• látky znečišťující vodu MeSH

We report the development of a novel method for detection of Bartonella DNA in ixodid ticks. The assay is based on a specific amplification of a part of 16S rRNA gene and 16S-23S rRNA intergenic spacer region of Bartonella sp. by nested PCR and Southern blot hybridization with specific DNA probe; the method is highly sensitive and specific. The screening of 327 unfed ticks collected in different urban and suburban areas of Czechia in 2003-2005 revealed the presence of Bartonella DNA in four Ixodes ricinus individuals (1.2%), two males, one female and one nymph.

• MeSH
• Bartonella genetika   izolace a purifikace   MeSH
• DNA bakterií genetika   MeSH
• intergenová DNA genetika   MeSH
• Ixodidae klasifikace   mikrobiologie   MeSH
• lidé MeSH
• oligonukleotidové sondy genetika   MeSH
• polymerázová řetězová reakce metody   MeSH
• ribozomální DNA genetika   MeSH
• RNA ribozomální 16S genetika   MeSH
• RNA ribozomální 23S genetika   MeSH
• senzitivita a specificita MeSH
• Southernův blotting metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• DNA bakterií MeSH
• intergenová DNA MeSH
• oligonukleotidové sondy MeSH
• ribozomální DNA MeSH
• RNA ribozomální 16S MeSH
• RNA ribozomální 23S MeSH

The genotype of Borrelia burgdorferi sensu lato was detected in 371 out of 1244 ticks. Borrelia determination was based on partial sequencing of the 16S rRNA gene and real-time polymerase chain reactions for identification and quantitation of ospA and recA genes. Different Borrelia spp. were identified; B. garinii in 40% ticks followed by B. afzelii (36.3%), B. burgdorferi sensu stricto (12.9%), B. valaisiana (3.5%), B. lusitaniae (0.8%), B. bissettii (0.5%) and B. miyamotoi-like (0.5%). Cultivation of 30 borrelia strains in BSK-H medium, among them B. valaisiana, B. bissettii-like and B. miyamotoi-like strains was unique in Czechia. Calibrated microfluidic-based quantification showed differences in the concentration of the nucleic acids and molar mass of the outer surface proteins of different Borrelia spp. with standard sensitivity and specificity and was helpful for their identification. The outer surface protein OspA was absent in B. miyamotoi-like and the OspB protein in B. valaisiana, B. lusitaniae and in three subtypes of B. garinii.

• MeSH
• antigeny povrchové chemie   genetika   MeSH
• bakteriální vakcíny chemie   genetika   MeSH
• Borrelia burgdorferi komplex genetika   izolace a purifikace   MeSH
• DNA bakterií chemie   genetika   MeSH
• fylogeneze MeSH
• genetická variace MeSH
• klíště mikrobiologie   MeSH
• lipoproteiny chemie   genetika   MeSH
• mikrofluidní analytické techniky MeSH
• molekulární sekvence - údaje MeSH
• polymerázová řetězová reakce MeSH
• proteiny vnější bakteriální membrány chemie   genetika   MeSH
• RecA-rekombinasy chemie   genetika   MeSH
• RNA ribozomální 16S chemie   genetika   MeSH
• sekvence nukleotidů MeSH
• sekvenční seřazení MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny povrchové MeSH
• bakteriální vakcíny MeSH
• DNA bakterií MeSH
• lipoproteiny MeSH
• OspA protein MeSH Prohlížeč
• proteiny vnější bakteriální membrány MeSH
• RecA-rekombinasy MeSH
• RNA ribozomální 16S MeSH

We have screened 91 migratory birds representing 32 species during the autumn of 2003 for the presence of the zoonotic pathogens Borrelia and Chlamydophila. Using polymerase chain reaction (PCR), B. burgdorferi sensu stricto was detected in cloacal swabs and, in two causes, also in throat swabs in 8 individuals (8.7 %) representing 7 birds species; B. garinii and B. afzelii were not detected. C. psittaci was detected only in cloacal swabs; 6 birds (6.6 %) from four species were found to be positive. The PCR products were sequenced and the sequences were compared phylogenetically with the gene sequences of 14 Chlamydophila strains retrieved from nucleotide databases; although the sequenced DNA was only 110 bp long, all obtained sequences created a new cluster with sublines branching from a position close to the periphery of the genus. All tested samples appear distinct within the known species and were most similar to C. felis or C. abortis.

• MeSH
• Borrelia burgdorferi klasifikace   genetika   izolace a purifikace   MeSH
• Chlamydophila psittaci klasifikace   genetika   izolace a purifikace   MeSH
• DNA bakterií analýza   chemie   genetika   MeSH
• farynx mikrobiologie   MeSH
• fylogeneze MeSH
• kloaka mikrobiologie   MeSH
• lymeská nemoc epidemiologie   mikrobiologie   veterinární   MeSH
• nemoci ptáků epidemiologie   mikrobiologie   MeSH
• polymerázová řetězová reakce MeSH
• psitakóza epidemiologie   mikrobiologie   veterinární   MeSH
• ptáci mikrobiologie   MeSH
• ribozomální DNA chemie   genetika   MeSH
• RNA ribozomální 16S genetika   MeSH
• sekvenční homologie nukleových kyselin MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Slovenská republika epidemiologie   MeSH
• Názvy látek
• DNA bakterií MeSH
• ribozomální DNA MeSH
• RNA ribozomální 16S MeSH

Selected cerebrospinal-fluid (CSF) parameters (intrathecal synthesis of Borrelia-specific antibodies, oligoclonal IgG bands, CSF-to-serum quotient of albumin as a marker of blood-CSF barrier function and cytology) and typical CSF profile in neuroborreliosis were evaluated with the aim of elucidating possible clinical and laboratory similarities of neuroborreliosis (NB) and other neurological diseases (OND). From the cohort of 58 patients (38 diagnosed for NB, 20 with OND) NB patients had positive Borrelia-specific IgG antibodies in 97 % and positive Borrelia-specific IgM antibodies in 55 %; oligoclonal IgG bands were detected in 55%. The blood-CSF barrier was impaired in 89%, positive cytology was detected in 97% of the NB patients. Evaluation of specific intrathecal synthesis improves CSF diagnosis of NB, therefore, a combined CSF analysis has to be considered along with the clinical picture and medical history when formulating the diagnosis of NB.

• MeSH
• Borrelia burgdorferi komplex * imunologie   MeSH
• diferenciální diagnóza MeSH
• hematoencefalická bariéra MeSH
• histocytochemie MeSH
• imunoglobulin G mozkomíšní mok   MeSH
• imunoglobulin M mozkomíšní mok   MeSH
• lidé MeSH
• lymská neuroborelióza mozkomíšní mok   diagnóza   MeSH
• mozkomíšní mok * chemie   cytologie   imunologie   MeSH
• nemoci nervového systému mozkomíšní mok   diagnóza   MeSH
• protilátky bakteriální mozkomíšní mok   MeSH
• sérový albumin mozkomíšní mok   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• imunoglobulin G MeSH
• imunoglobulin M MeSH
• protilátky bakteriální MeSH
• sérový albumin MeSH