Článek se zaměřuje na rutinně nekultivovatelné treponemové infekce, které i nadále představují celosvětový problém. Pojednává o současných poznatcích v oblasti molekulární genetiky a zdůrazňuje důležitost implementace aktuálních výzkumů do dermatovenerologie. Soustředí se na využití metody PCR v diagnostice a propojení klinické medicínské praxe s vědeckými poznatky v oblasti molekulární genetiky u treponemových infekcí.
The article deals with routinely uncultivable treponemal infections that continue to be a global problem. The current knowledge in the field of molecular genetics is presented and the importance of implementing current research into dermatovenerology is emphasized. The article focuses on the use of the PCR method in diagnosis as well as on linking clinical medical practice with scientific knowledge in the field of molecular genetics in treponemal infections.
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.
- 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 MeSH
Syphilis is a multistage sexually transmitted disease caused by Treponema pallidum ssp. pallidum. In the Czech Republic, there are around 700-800 new syphilis cases annually, continuously increasing since 2012. This study analyzed a total of 1228 samples from 2004 to 2022. Of the PCR-positive typeable samples (n = 415), 68.7% were fully-typed (FT), and 31.3% were partially-typed. Most of the identified isolates belonged to the SS14-clade and only 6.3% were the Nichols-like cluster. While in the beginning of sample collection isolates have been macrolide-susceptible, recent isolates are completely resistant to macrolides. Among the FT samples, 34 different allelic profiles (APs) were found. Most of the profiles (n = 27) appeared just once in the Czech population, while seven profiles were detected more than twice. The most frequent APs belonged to two separate groups of SS14-like isolates, including group of 1.3.1 (ST 1) and 1.26.1 (ST 25) profiles, and the second group containing 1.1.8 (ST 3), 1.1.1 (ST 2), and 1.1.3 (ST 11) (representing 57.5%, and 25.3% of all detected APs, respectively). Both groups consistently differed in 6 nucleotide positions in five genes (TP0150, TP0324, TP0515, TP0548, and TP0691) coding amino-acid replacements suggesting that one or more of these differences could be involved in the higher success of the first group.
- MeSH
- alely * MeSH
- dospělí MeSH
- genotyp MeSH
- lidé středního věku MeSH
- lidé MeSH
- makrolidy farmakologie MeSH
- multilokusová sekvenční typizace * MeSH
- syfilis * mikrobiologie epidemiologie genetika MeSH
- Treponema pallidum * genetika izolace a purifikace MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Globally, 94% of Treponema pallidum subsp. pallidum (TPA) clinical strains belong to the SS14-like group and 6% to the Nichols-like group, with a prevalence of macrolide resistance of 90%. Our goal was to determine whether local TPA strain distribution and macrolide resistance frequency have changed significantly since our last report, which revealed that Buenos Aires had a high frequency of Nichols-like strains (27%) and low levels of macrolide resistance (14%). Swab samples from patients with suspected syphilis were collected during 2015-2019 and loci TP0136, TP0548, TP0705 were sequenced in order to perform multilocus sequence typing. Strains were classified as Nichols-like or SS14-like. The presence of macrolide resistance-associated mutations was determined by examination of the 23S rDNA gene sequence. Of 46 typeable samples, 37% were classified as Nichols-like and 63% as SS14-like. Macrolide resistance prevalence was 45.7%. Seven allelic profiles were found, five were SS14-like and two were Nichols-like. The frequency of Nichols-like strains increased between studies (26.8% vs. 37%, p = 0.36). A dramatic increase was found in the frequency of macrolide resistant strains between studies (14.3% vs. 45.7%, p = 0.005). Our results are in agreement with international trends and underscore the need to pursue further TPA molecular typing studies in South America.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence genetika MeSH
- infekce bakteriemi rodu Treponema * MeSH
- lidé MeSH
- makrolidy farmakologie MeSH
- multilokusová sekvenční typizace MeSH
- ribozomální DNA MeSH
- Treponema pallidum * genetika MeSH
- Treponema MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Argentina MeSH
Bejel (endemic syphilis) is a neglected non-venereal disease caused by Treponema pallidum subsp. endemicum (TEN). Although it is mostly present in hot, dry climates, a few cases have been found outside of these areas. The aim of this work was the sequencing and analysis of TEN isolates obtained from "syphilis patients" in Cuba, which is not considered an endemic area for bejel. Genomes were obtained by pool segment genome sequencing or direct sequencing methods, and the bioinformatics analysis was performed according to an established pipeline. We obtained four genomes with 100%, 81.7%, 52.6%, and 21.1% breadth of coverage, respectively. The sequenced genomes revealed a non-clonal character, with nucleotide variability ranging between 0.2-10.3 nucleotide substitutions per 100 kbp among the TEN isolates. Nucleotide changes affected 27 genes, and the analysis of the completely sequenced genome also showed a recombination event between tprC and tprI, in TP0488 as well as in the intergenic region between TP0127-TP0129. Despite limitations in the quality of samples affecting breadth of sequencing coverage, the determined non-clonal character of the isolates suggests a persistent infection in the Cuban population rather than a single outbreak caused by imported case.
Cílem studie bylo vyhodnotit klinická, epidemiologická a demografická data pacientů se syfilis léčených na Dermatovenerologické klinice 1. LF UK a Všeobecné fakultní nemocnice v Praze (STD centrum) v roce 2017–2019 a porovnat je s předchozím průzkumem provedeným v roce 2007–2009, s celorepublikovými údaji z Registru pohlavních nemocí a s daty evropské databáze (The European Surveillance System of the ECDC). V průběhu let 2017–2019 jsme hospitalizovali 325 pacientů se syfilis, 83,4 % mužů a 16,6 % žen. Celkový počet pacientů se snížil o 148 případů (31,3 %), výraznější je pokles u žen (61,2 %) než u mužů (18,9 %). Nárůst poměru mužů vůči ženám je statisticky významný (p < 0,001). Podíl cizinců byl 26,5 % a signifikantně se nezměnil. V souboru mužů klesl podíl primární syfilis (24,4 % vs. 32,0 %; p = 0,046) a došlo k významnému nárůstu počtu HIV pozitivních mužů (7,7 % vs. 3,0 %; p = 0,017) při zachování přibližně stejného podílu homosexuálních mužů (p = 0,934). Podíl homo/bisexuálních mužů činil 48,4 % ze souboru všech pacientů a 58,3 % ze souboru mužů. V souboru žen byl tradičně zaznamenán nízký počet případů primární a sekundární syfilis, tento podíl se statisticky významně nezměnil. Rovněž se signifikantně nezměnil podíl gravidních žen (p = 0,371). Vyšetření vzorků na makrolidovou rezistenci ukázalo podstatné snížení počtu senzitivních vzorků pro období 2017–2019 (9,5 % vs. 57,6 %). Pro analýzu trendů a cílenou prevenci syfilis je nutný přesný a průběžný sběr epidemiologických a klinických dat. Z našich i celoevropských údajů vyplývá, že nejohroženější skupinou jsou muži mající sex s muži. Pravidelný screening na syfilis a ostatní sexuálně přenosné infekce musí být u této skupiny prioritou bez ohledu na HIV status.
The aim of the study was to evaluate clinical, epidemiological and demographic data of patients treated with syphilis at the Department of Dermatovenereology of the 1st faculty of medicine and General University Hospital in Prague (STD centre) in 2017–2019 and compare them with the previous survey carried out in 2007–2009 and with the nationwide data from the Czech Registry of veneral diseases and the european data from the European Surveillance System of the ECDC. A total of 325 patients (83,4 % men and 16,6 % women) were hospitalized during the period 2017–2019. The total number of hospitalised patients decreased by 148 cases (31.3 %), the decrease was more pronounced in the female group (61.2%) as compared to the male group (18.9%). The increase of the male-to-female ratio is statistically significant (p < 0.001). The patients’ age has not changed in the long term, most of them were aged between 30 and 40. The percentage of foreigners was 26.5% and did not change significantly. In the male group, the proportion of primary syphilis decreased (24.4% vs 32.0 %; p = 0.046) and there was a significant increase in the number of HIV-positive patients (7.7% vs 3.0 %; p = 0.017), while the proportion of homo/bisexual men remained approximately the same (p = 0.934). The proportion of homo/bisexual men was 48.4% of all patients and 58.3% of men. In the female group, the number of primary and secondary syphilis cases was low and this proportion did not change significantly. Similarly, the proportion of pregnant women did not change significantly either (p = 0.371). The testing of samples for macrolide resistance showed a significant decrease in the number of sensitive strains (9.5% vs 57.6%). Accurate and continuous collection of epidemiological and clinical data is necessary for trend analysis and targeted syphilis prevention. Our as well as European data show that men having sex with men are the most vulnerable group. Regular screening for syphilis and other sexually transmitted infections must be a priority in this group, regardless of the HIV status.
- Klíčová slova
- alelické profily,
- MeSH
- epidemiologické studie MeSH
- lidé MeSH
- sexuálně přenosné nemoci epidemiologie přenos MeSH
- syfilis * epidemiologie MeSH
- Treponema pallidum MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví 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
BACKGROUND: Treponema pallidum subspecies pallidum (TPA) and subsp. endemicum (TEN) are the causative agents of syphilis and bejel, respectively. TEN shows similar clinical manifestations and is morphologically and serologically indistinguishable from TPA. Recently, bejel was found outside of its assumed endemic areas. Using molecular typing we aimed to discover bejel and characterize circulating TPA types among syphilis cases with Surinamese, Antillean and Dutch ethnicity in Amsterdam. METHODS: DNA was extracted from 137 ulcer swabs, which tested positive in the in-house diagnostic PCR targeting the polA gene. Samples were collected between 2006 and 2018 from Surinamese, Antillean and Dutch patients attending the Amsterdam STI clinic. Multilocus sequence typing was performed by partial sequence analysis of the tp0136, tp0548 and tp0705 genes. In addition, the 23S rRNA loci were analyzed for A2058G and A2059G macrolide resistance mutations. RESULTS: We found 17 distinct allelic profiles in 103/137 (75%) fully typed samples, which were all TPA and none TEN. Of the strains, 82.5% were SS14-like and 17.5% Nichols-like. The prevalence of Nichols-like strains found in this study is relatively high compared to nearby countries. The most prevalent types were 1.3.1 (42%) and 1.1.1 (19%), in concordance with similar TPA typing studies. The majority of the TPA types found were unique per country. New allelic types (7) and profiles (10) were found. The successfully sequenced 23S rRNA loci from 123/137 (90%) samples showed the presence of 79% A2058G and 2% A2059G mutations. CONCLUSIONS: No TEN was found in the samples from different ethnicities residing in Amsterdam, the Netherlands, so no misdiagnoses occurred. Bejel has thus not (yet) spread as a sexually transmitted disease in the Netherlands. The strain diversity found in this study reflects the local male STI clinic population which is a diverse, mixed group.
- MeSH
- alely MeSH
- bakteriální geny * MeSH
- dospělí MeSH
- etnicita statistika a číselné údaje MeSH
- lidé MeSH
- syfilis epidemiologie etnologie mikrobiologie MeSH
- taxonomické DNA čárové kódování MeSH
- Treponema pallidum klasifikace genetika patogenita MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Nizozemské Antily MeSH
- Nizozemsko MeSH
- Surinam 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.
- 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 MeSH
