A set of 415 clinical samples isolated from 294 patients suspected of having syphilis collected in the Czech Republic between 2004 and 2010 was tested for the presence of treponemal DNA. Standard serological tests showed that 197 patients were syphilis-seropositive and 97 patients were syphilis-seronegative. In each sample, PCR tests for polA (TP0105), tmpC (TP0319), TP0136, TP0548 and 23S rRNA genes were performed. Samples taken from 91 patients were PCR-positive. Molecular typing of treponemal DNA was based on the sequencing of TP0136, TP0548 and 23S rRNA genes. Treponemal DNA was typeable in samples taken from 64 PCR-positive patients and 9 different genotypes were found. The proportion of treponemal strains resistant to macrolide antibiotics was 37.3%. In the DNA samples taken from 39 patients, a parallel treponemal typing approved by Centers for Disease Control and Prevention was performed. The variants of arp and tpr genes appear to combine independently with sequence variants of TP0136, TP0548 and 23S rRNA genes.

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Resolving the taxonomy of emerging zoonotic pathogens in the Trichophyton benhamiae complex

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Over a 12-year period, from 1987 to 1998, the spectrum of aetiological agents isolated from 11208 patients (6265 females and 4943 males) suspected of having dermatomycosis was analysed in the mycological laboratory of the Department of Dermatology, Charles University, Prague. The most frequently examined locations were toenails (34.9%), feet (15.6%), fingernails (12.8%), toe webs (11%), trunk (10%) and hands (8.7%). Dermatophytes were isolated from 5605 (30.2%) of all 18528 samples examined. Among dermatophytes, the most common infectious agent was Trichophyton rubrum (90.2%) followed by Trichophyton mentagrophytes (6.6%), Microsporum canis (1.8%) and Epidermophyton floccosum (1.4%). Trichophyton verrucosum, Microsporum persicolor and Microsporum gypseum were rare. During this 12-year period the pattern of aetiological agents of dermatomycoses in Prague was relatively stable.

BackgroundMolecular typing of syphilis-causing strains provides important epidemiologic data. We tested whether identified molecular subtypes were identical in PCR-positive parallel samples taken from the same patient at a same time. We also tested whether subtype prevalence differs in skin and blood samples.ResultsEighteen syphilis positive patients (showing both positive serology and PCR), with two PCR-typeable parallel samples taken at the same time, were tested with both CDC (Centers for Disease Control and Prevention) and sequence-based typing. Samples taken from 9 of 18 patients were completely typed for TP0136, TP0548, 23S rDNA, arp, and tpr loci. The CDC typing revealed 11 distinct genotypes while the sequence-based typing identified 6 genotypes. When results from molecular typing of TP0136, TP0548, and 23S rDNA were analyzed in samples taken from the same patient, no discrepancies in the identified genotypes were found; however, there were discrepancies in 11 of 18 patients (61.1%) samples relative to the arp and tpr loci. In addition to the above described typing, 127 PCR-positive swabs and whole blood samples were tested for individual genotype frequencies. The repetition number for the arp gene was lower in whole blood (WB) samples compared to swab samples. Similarly, the most common tpr RFLP type “d” was found to have lower occurrence rates in WB samples while type “e” had an increased occurrence in these samples.ConclusionsDifferences in the CDC subtypes identified in parallel samples indicated genetic instability of the arp and tpr loci and suggested limited applicability of the CDC typing system in epidemiological studies. Differences in treponemal genotypes detected in whole blood and swab samples suggested important differences between both compartments and/or differences in adherence of treponeme variants to human cells.

SUMMARYThe aim of the study was to evaluate clinical, epidemiological and demographic data of patients with syphilis hospitalized at the Department of Dermatovenereology of the General Teaching Hospital in 2009 and to identify the groups at high risk of sexually transmitted diseases. The results were compared with the previous surveys carried out between 1999 and 2005 and also with the data of the National Registry of Venereal Diseases for 2008.A total of 232 patients were hospitalized in 2009 (including 26 women admitted for compulsory retreatment in pregnancy). We noticed a 25% increase in the number of patients in comparison with the year 2008. Of the total number of patients, 206 were patients with newly diagnosed syphilis, of which 153 (74.3%) were men and 53 (25.7%) women. There was 22.3% of patients with primary and 31.6% with secondary syphilis. As in the previous years, heterosexual contact remained the most frequent route of syphilis transmission. However, the number of homosexual and bisexual men increased, amounting to 64.7% of all hospitalized men. Four men were HIV positive. The age distribution did not change; majority of the patients were aged between 30 and 40. The proportion of foreigners also remained unchanged (19.9%); they came mainly from the Slovak Republic (31.1%) and Ukraine (26.7%). The absolute number of reported syphilis cases has been increasing in the Czech Republic since 2006. From the epidemiologic point of view, the increasing incidence of early stages of the disease is alarming. Men prevail over women among the infected persons; the men having sex with men (MSM) has been increasing in the last three years.

Onychomycosis is one of the most common nail disorders. Its current treatment is not satisfactorily effective and often causes adverse side effects. This study aims to determine the optimal conditions for non-thermal plasma (NTP) inactivation of the most common dermatophytes in vitro and to apply it in patient`s therapy. The in vitro exposure to NTP produced by negative DC corona discharge caused full inactivation of Trichophyton spp. if applied during the early growth phases. This effect decreased to negligible inactivation with the exposure applied six days after inoculation. In a group of 40 patients with onychomycosis, NTP therapy was combined with nail plate abrasion and refreshment (NPAR) or treatment with antimycotics. The cohort included 17 patients treated with NPAR combined with NTP, 11 patients treated with antimycotics and NTP, and 12 patients treated with NPAR alone. The combination of NPAR and NTP resulted in clinical cure in more than 70% of patients. The synergistic effect of NPAR and NTP caused 85.7% improvement of mycological cure confirmed by negative microscopy and culture of the affected nail plate. We conclude that NTP can significantly improve the treatment of onychomycosis.