• Klíčová slova
• TREPONEMAL INFECTIONS/immunology *,
• MeSH
• infekce bakteriemi rodu Treponema imunologie   MeSH
• lidé MeSH
• protilátky * MeSH
• Treponema pallidum * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• protilátky * MeSH

• MeSH
• infekce bakteriemi rodu Treponema diagnóza   MeSH
• lidé MeSH
• muramidasa * MeSH
• testy imobilizace Treponema pallidum * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• muramidasa * MeSH

Syphilis is a multistage sexually transmitted disease caused by Treponema pallidum ssp. pallidum (TPA). This study analyzed clinical samples collected from patients with a diagnosed syphilis infection from 2004-2022, isolated in the Czech Republic. Mucocutaneous swab samples (n = 543) from 543 patients were analyzed, and from these samples, 80.11 % (n = 435) were PCR positive, and 19.89 % (n = 108) were PCR negative for TPA DNA. Swabs were more often positive when collected from syphilis patients in the primary and secondary stages, compared to the latent or unknown stage. There was no significant difference in PCR positivity between the primary and secondary stages (p = 0.099). In IgM-positive patients, a statistically significant association with PCR-positivity was found in samples from seropositive (p = 0.033) and serodiscrepant (RPR negative) patients (p = 0.0006). When assessing our laboratory-defined cases of syphilis, the RPR, IgM, and PCR tests were similarly effective (within the range of 80.1-86.1 %). However, parallel testing with these methods was even more effective, i.e., RPR + PCR was 96.1 % effective and RPR + IgM + PCR was 97.8 % effective. A combination of RPR + PCR, or a combination of all three tests (RPR, IgM, and PCR) can therefore be used to reliably detect active syphilis cases, including reinfections. Our findings show that the reverse algorithm for detecting syphilis could be substantially improved by adding IgM and PCR testing.

• Klíčová slova
• Nontreponemal tests, PCR syphilis detection, RPR, Syphilis, Syphilis diagnostics, Syphilis serology, Treponemal tests,
• MeSH
• DNA bakterií genetika   MeSH
• dospělí MeSH
• imunoglobulin M * krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• polymerázová řetězová reakce * metody   MeSH
• protilátky bakteriální krev   MeSH
• senzitivita a specificita MeSH
• sérologická diagnostika syfilis metody   MeSH
• syfilis * diagnóza   mikrobiologie   MeSH
• Treponema pallidum * genetika   izolace a purifikace   imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• DNA bakterií MeSH
• imunoglobulin M * MeSH
• protilátky bakteriální MeSH

BACKGROUND: The aim of this study was to quantify the seroprevalence of hare treponematosis in European brown hare (Lepus europaeus) populations in the Czech Republic and to test for an association between treponematosis prevalence and the altitude of the areas in which hares were sampled. We tested 289 serum samples of brown hares collected between 2015 and 2017. The sampling areas included 12 districts (73 villages) distributed throughout the Czech Republic. Serum samples were tested for the presence of antibodies against the causative agent of hare treponematosis (Treponema paraluisleporidarum ecovar Lepus, TPeL) using two serological tests for human syphilis that cross-react with TPeL: the Treponema pallidum hemagglutination assay (TPHA) and the fluorescent treponemal antibody absorption (FTA-ABS) test. To account for the imperfect diagnostic sensitivity and specificity of each test, apparent prevalence estimates of TPeL were converted to true prevalence estimates using the Rogan Gladen estimator. The correlation between TPeL true seroprevalence and altitude of sampling areas was analyzed using Pearson's correlation coefficient at three levels of spatial resolution: (1) four groups, each composed of two merged districts, with ≥20 samples collected, differing in their altitude median (206, 348, 495, and 522 m above sea level); (2) separately tested eight districts, where ≥20 samples were collected per district; and (3) 27 groups composed of villages of the same altitude level distributed across the whole dataset. RESULTS: One hundred and seven of the 289 samples were seropositive to both tests, the FTA-ABS test was positive for an additional 47 samples. Seropositive samples were found in all 12 districts. True seroprevalence of TPeL in the sampled hares was 52% (95% confidence interval 46 to 58%). A statistically significant negative correlation between TPeL seroprevalence and altitude was identified at the district level (Pearson's r = - 0.722, p = 0.043). CONCLUSIONS: Between 2015 and 2017 hare treponematosis was present at a relatively high prevalence in brown hares in all 12 districts in the Czech Republic where sampling was carried out. The seroprevalence of TPeL in brown hares was negatively correlated with the altitude of the areas in which hares were sampled.

• Klíčová slova
• Game animals, Hare disease, Lagomorphs, Lepus europaeus, Lepus timidus, Lesion, Treponema paraluisleporidarum, Treponematosis, Venereal disease, Wildlife disease,
• MeSH
• infekce bakteriemi rodu Treponema epidemiologie   veterinární   MeSH
• nadmořská výška MeSH
• séroepidemiologické studie MeSH
• zajíci * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

Syphilis, caused by Treponema pallidum ssp. pallidum (TPA), is a persisting global health problem. Although syphilis diagnostics relies mainly on serology, serological tests have some limitations, and it is recommended that the final diagnosis be supported by additional tests. The purpose of this study was to analyze the relationship between serology and PCR in syphilis diagnostics. From the year 2004 to May 2019, a total of 941 samples were taken from 833 patients suspected of having syphilis, in Czech Republic. In all these samples, both nested PCR detection of TPA and serology testing were performed. Of the 941 samples, 126 were seronegative, 651 were seropositive, and 164 were serodiscrepant. Among seronegative samples (n = 126), 11 were PCR-positive (8.7%). Among seropositive samples (n = 651; i.e., samples positive for both non-treponemal and treponemal serology tests), 368 samples were PCR-positive (56.5%). The remaining 164 serodiscrepant samples included RPR negative and treponemal serological test-positive samples (n = 154) and a set of 10 RPR-positive samples negative in treponemal serological tests. While the first group revealed 73 PCR-positive samples (47.4%), the second revealed 5 PCR positive samples (50.0%). PCR detection rates were highest in primary syphilis, with lower rates in the secondary and undetermined syphilis stages. As shown here, the nested PCR can improve diagnostics of syphilis, especially in seronegative patients and in patients with discrepant serology.

• MeSH
• lidé MeSH
• polymerázová řetězová reakce * MeSH
• retrospektivní studie MeSH
• sérologická diagnostika syfilis metody   MeSH
• syfilis krev   diagnóza   MeSH
• Treponema genetika   imunologie   izolace a purifikace   fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Using the Captia Select Syph-G enzyme-linked immunosorbent assay (ELISA), we analyzed the sera of 1,771 patients from a high-risk population attending sexually transmitted disease (STD) clinics. We focused on discrepancies between the results of the immunoglobulin G (IgG) ELISA and the Treponema pallidum hemagglutination (TPHA) test. We identified 57 patients (3.22%) with conflicting results in the IgG ELISA and TPHA test. In order to resolve these discrepancies, these patients' health records were reviewed and additional serological tests (rapid plasma reagin, IgM ELISA, fluorescent treponemal antibody absorption, and Western blotting) were performed. We subsequently diagnosed 22 of these 57 (38.6%) patients with late latent syphilis. None of the patients with discordant test results was diagnosed with early syphilis. We followed 35 of these 57 patients, analyzing two consecutive serum samples at 3 weeks and at 3 months. Discordant results persisted in 12 (33.3%) patients. We successfully resolved the test result discrepancies for 28 patients (80%) involved in follow-up. Captia SelectSyph-G ELISA showed a sensitivity of 99.0%, a specificity of 98.0%, and positive and negative predictive values of 99.3% and 97.2%, respectively. Based on the results of this study, we conclude that the Captia SelectSyph-G ELISA is a reliable tool for syphilis testing in a high-risk population and recommend the utilization of the Captia SelectSyph-G ELISA as a confirmatory test in at-risk patients.

• MeSH
• dítě MeSH
• dospělí MeSH
• ELISA MeSH
• hemaglutinační testy MeSH
• imunoglobulin G krev   MeSH
• lidé MeSH
• prediktivní hodnota testů MeSH
• protilátky bakteriální krev   MeSH
• reagenční diagnostické soupravy * MeSH
• reprodukovatelnost výsledků MeSH
• rizikové faktory MeSH
• senzitivita a specificita MeSH
• sérologická diagnostika syfilis MeSH
• syfilis diagnóza   MeSH
• těhotenství MeSH
• Treponema pallidum imunologie   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• imunoglobulin G MeSH
• protilátky bakteriální MeSH
• reagenční diagnostické soupravy * MeSH

Although the link between microbial infections and Alzheimer's disease (AD) has been demonstrated in multiple studies, the involvement of pathogens in the development of AD remains unclear. Here, we investigated the frequency of the 10 most commonly cited viral (HSV-1, EBV, HHV-6, HHV-7, and CMV) and bacterial (Chlamydia pneumoniae, Helicobacter pylori, Borrelia burgdorferi, Porphyromonas gingivalis, and Treponema spp.) pathogens in serum, cerebrospinal fluid (CSF) and brain tissues of AD patients. We have used an in-house multiplex PCR kit for simultaneous detection of five bacterial and five viral pathogens in serum and CSF samples from 50 AD patients and 53 healthy controls (CTRL). We observed a significantly higher frequency rate of AD patients who tested positive for Treponema spp. compared to controls (AD: 62.2 %; CTRL: 30.3 %; p-value = 0.007). Furthermore, we confirmed a significantly higher occurrence of cases with two or more simultaneous infections in AD patients compared to controls (AD: 24 %; CTRL 7.5 %; p-value = 0.029). The studied pathogens were detected with comparable frequency in serum and CSF. In contrast, Borrelia burgdorferi, human herpesvirus 7, and human cytomegalovirus were not detected in any of the studied samples. This study provides further evidence of the association between microbial infections and AD and shows that paralleled analysis of multiple sample specimens provides complementary information and is advisable for future studies.

• Klíčová slova
• Alzheimer's disease, Microbial infection, PCR, Pathogen, Serum, Treponema spp.,
• MeSH
• Alzheimerova nemoc * epidemiologie   mikrobiologie   MeSH
• infekce bakteriemi rodu Treponema * epidemiologie   MeSH
• lidé MeSH
• lidský herpesvirus 6 MeSH
• studie případů a kontrol MeSH
• Treponema * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Pathogenic treponemes related to Treponema pallidum are both human (causing syphilis, yaws, bejel) and animal pathogens (infections of primates, venereal spirochetosis in rabbits). A set of 11 treponemal genome sequences including those of five Treponema pallidum ssp. pallidum (TPA) strains (Nichols, DAL-1, Mexico A, SS14, Chicago), four T. p. ssp. pertenue (TPE) strains (CDC-2, Gauthier, Samoa D, Fribourg-Blanc), one T. p. ssp. endemicum (TEN) strain (Bosnia A) and one strain (Cuniculi A) of Treponema paraluisleporidarum ecovar Cuniculus (TPeC) were tested for the presence of positively selected genes. METHODOLOGY/PRINCIPAL FINDINGS: A total of 1068 orthologous genes annotated in all 11 genomes were tested for the presence of positively selected genes using both site and branch-site models with CODEML (PAML package). Subsequent analyses with sequences obtained from 62 treponemal draft genomes were used for the identification of positively selected amino acid positions. Synthetic biotinylated peptides were designed to cover positively selected protein regions and these peptides were tested for reactivity with the patient's syphilis sera. Altogether, 22 positively selected genes were identified in the TP genomes and TPA sets of positively selected genes differed from TPE genes. While genetic variability among TPA strains was predominantly present in a number of genetic loci, genetic variability within TPE and TEN strains was distributed more equally along the chromosome. Several syphilitic sera were shown to react with some peptides derived from the protein sequences evolving under positive selection. CONCLUSIONS/SIGNIFICANCE: The syphilis-, yaws-, and bejel-causing strains differed relative to sets of positively selected genes. Most of the positively selected chromosomal loci were identified among the TPA treponemes. The local accumulation of genetic variability suggests that the diversification of TPA strains took place predominantly in a limited number of genomic regions compared to the more dispersed genetic diversity differentiating TPE and TEN strains. The identification of positively selected sites in tpr genes and genes encoding outer membrane proteins suggests their role during infection of human and animal hosts. The driving force for adaptive evolution at these loci thus appears to be the host immune response as supported by observed reactivity of syphilitic sera with some peptides derived from protein sequences showing adaptive evolution.

• MeSH
• bakteriální geny * MeSH
• biologická adaptace * MeSH
• dospělí MeSH
• genomika MeSH
• genotyp * MeSH
• lidé MeSH
• mladý dospělý MeSH
• selekce (genetika) MeSH
• syfilis mikrobiologie   patologie   MeSH
• Treponema pallidum klasifikace   genetika   izolace a purifikace   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Molecular 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. RESULTS: Eighteen 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. CONCLUSIONS: Differences 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.

• MeSH
• bakteriální proteiny genetika   MeSH
• genetická variace * MeSH
• krev mikrobiologie   MeSH
• kůže mikrobiologie   MeSH
• lidé MeSH
• molekulární typizace metody   MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• RNA ribozomální 23S genetika   MeSH
• syfilis mikrobiologie   MeSH
• Treponema pallidum klasifikace   genetika   izolace a purifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• bakteriální proteiny MeSH
• RNA ribozomální 23S MeSH

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.

• MeSH
• antibakteriální látky terapeutické užití   MeSH
• bakteriální léková rezistence genetika   MeSH
• DNA bakterií analýza   MeSH
• dospělí MeSH
• fenotyp MeSH
• genetická variace MeSH
• genotyp MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• polymerázová řetězová reakce MeSH
• ribotypizace * metody   MeSH
• RNA ribozomální 23S genetika   MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA MeSH
• syfilis diagnóza   farmakoterapie   epidemiologie   mikrobiologie   MeSH
• Treponema pallidum účinky léků   genetika   imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• antibakteriální látky MeSH
• DNA bakterií MeSH
• RNA ribozomální 23S MeSH