OBJECTIVES: Treponema pallidum subsp. pallidum (T. pallidum) is the etiological agent of syphilis, a sexually transmitted disease of global public health importance. The objective of this study was to introduce a novel in vitro protocol for isolation of T. pallidum directly from patients' clinical samples, eliminating the need for rabbit propagation. METHODS: Four oral and five genital swabs were collected from nine epidemiologically unrelated patients at two hospitals in Brno, Czech Republic. Swabs were submerged in TpCM-2 medium for transport. Samples were then placed on a 0.4 μm filters and incubated for 2.5 hours. During this period, spiral T. pallidum cells passed through the filter pores to the well containing TpCM-2 medium and rabbit feeder cells (Sf1Ep). Stable T. pallidum cultures (containing >1 × 107 treponemes) were achieved by subculturing every 7 days into fresh well. RESULTS: A successful protocol for in vitro isolation of T. pallidum was established. From the nine clinical specimens processed, six T. pallidum cultures (MU1-MU6) were derived after 14 to 112 days of cultivation. Five of these strains (MU1-MU5) belonged to SS14-like cluster and shared the same allelic profile 1.3.1. The remaining strain (MU6) was identified as a Nichols-like strain with an allelic profile 9.16.3. DISCUSSION: The introduced in vitro protocol enables isolation of T. pallidum from clinical material, including frozen samples, without the need for experimental rabbits. This method facilitates the isolation of contemporary, clinically relevant treponemal strains.
- MeSH
- Bacteriological Techniques * methods MeSH
- Rabbits MeSH
- Humans MeSH
- Genitalia microbiology MeSH
- Syphilis * microbiology diagnosis MeSH
- Treponema pallidum * isolation & purification genetics classification MeSH
- Mouth microbiology MeSH
- Animals MeSH
- Check Tag
- Rabbits MeSH
- Humans MeSH
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
Mezi celosvětově rozšířené sexuálně přenosné nemoci je řazeno infekční onemocnění syfilis. Kvůli neuroinvazivitě infikujícího kmene treponem se u pacientů můžeme setkat s velmi pestrým postižením centrální nervové soustavy, k jehož projevům může dojít během kteréhokoli stadia onemocnění. Článek obsahuje přehled jednotlivých stadií syfilis a neurosyfilis. Formou kazuistického sdělení mladého pacienta popisujeme terciální stadium syfilis pod obrazem progresivní paralýzy.
The infectious disease syphilis is among the most common sexually transmitted diseases worldwide. Due to the neuroinvasiveness of the infecting treponemal strain, patients can encounter a wide variety of central nervous system involvement, which can occur during any stage of the disease. This article reviews the different stages of syphilis and neurosyphilis. In the form of case report of a young patient, we describe the late stage of syphilis as general paresis.
- MeSH
- Diagnosis, Differential MeSH
- Adult MeSH
- Case Reports as Topic MeSH
- Cognitive Dysfunction diagnosis etiology microbiology MeSH
- Humans MeSH
- Neurologic Manifestations MeSH
- Neurosyphilis * diagnosis epidemiology complications MeSH
- Sexually Transmitted Diseases diagnosis drug therapy classification MeSH
- Treponema pathogenicity MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Male MeSH
BACKGROUND: Treponema pallidum subspecies pertenue (TPE) is the causative agent of human and nonhuman primate (NHP) yaws infection. The discovery of yaws bacterium in wild populations of NHPs opened the question of transmission mechanisms within NHPs, and this work aims to take a closer look at the transmission of the disease. METHODOLOGY/PRINCIPAL FINDINGS: Our study determined eleven whole TPE genomes from NHP isolates collected from three national parks in Tanzania: Lake Manyara National Park (NP), Serengeti NP, and Ruaha NP. The bacteria were isolated from four species of NHPs: Chlorocebus pygerythrus (vervet monkey), Cercopithecus mitis (blue monkey), Papio anubis (olive baboon), and Papio cynocephalus (yellow baboon). Combined with previously generated genomes of TPE originating from NHPs in Tanzania (n = 11), 22 whole-genome TPE sequences have now been analyzed. Out of 231 possible combinations of genome-to-genome comparisons, five revealed an unexpectedly high degree of genetic similarity in samples collected from different NHP species, consistent with inter-species transmission of TPE among NHPs. We estimated a substitution rate of TPE of NHP origin, ranging between 1.77 × 10-7 and 3.43 × 10-7 per genomic site per year. CONCLUSIONS/SIGNIFICANCE: The model estimations predicted that the inter-species transmission happened recently, within decades, roughly in an order of magnitude shorter time compared to time needed for the natural diversification of all tested TPE of Tanzanian NHP origin. Moreover, the geographical separation of the sampling sites (NPs) does not preclude TPE transmission between and within NHP species.
- MeSH
- Chlorocebus aethiops MeSH
- Cercopithecus microbiology MeSH
- Yaws * microbiology transmission MeSH
- Phylogeny * MeSH
- Genome, Bacterial MeSH
- Humans MeSH
- Monkey Diseases microbiology transmission MeSH
- Papio anubis microbiology MeSH
- Papio cynocephalus microbiology genetics MeSH
- Primates microbiology MeSH
- Whole Genome Sequencing * MeSH
- Treponema pallidum genetics isolation & purification classification MeSH
- Treponema MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Tanzania MeSH
Syfilis je systémové, chronické a celosvětově rozšířené infekční onemocnění způsobené bakterií Treponema pallidum, jejím jediným přirozeným hostitelem je člověk. K přenosu infekce dochází převážně pohlavním stykem (90 %), dále je však možný i přenos transplacentární nebo přenos krevní cestou, jiné cesty nákazy bývají vzácné. Závažnost syfilis je dána rizikem vzniku postižení různých orgánů, zejména kůže, oka, CNS, kardiovaskulárního a muskuloskeletálního systému a rizikem přenosu infekce na plod během gravidity či porodu u neléčené matky. Článek se zaměřuje na předání ucelených a stručných informací o získané syfilis (lat. syphilis acquisita) u dospělých.
Syphilis is a systemic, chronic, and worldwide spread infectious disease caused by the bacterium Treponema pallidum, and its only natural host is man. The disease is transmitted predominantly through sexual contact (90%), but transplacental infection and blood transmission are also possible; other routes of infection are rare. The severity of syphilis is determined by the risk of various organs, particularly the skin, eye, CNS, cardiovascular and musculoskeletal systems and the risk of transmission to the foetus during pregnancy or birth in an untreated mother. The article aims to provide comprehensive and concise information on acquired syphilis (lat. syphilis acquisita) in adults.
- MeSH
- Ceftriaxone administration & dosage MeSH
- Diagnosis, Differential MeSH
- Adult MeSH
- Doxycycline administration & dosage MeSH
- Skin Manifestations MeSH
- Humans MeSH
- Neurosyphilis pathology MeSH
- Penicillins administration & dosage MeSH
- Syphilis Serodiagnosis classification MeSH
- Practice Guidelines as Topic MeSH
- Severity of Illness Index MeSH
- Syphilis * diagnosis epidemiology drug therapy pathology MeSH
- Treponema pallidum classification pathogenicity MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Publication type
- Review 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.
- MeSH
- DNA, Bacterial genetics MeSH
- Adult MeSH
- Immunoglobulin M * blood MeSH
- Middle Aged MeSH
- Humans MeSH
- Young Adult MeSH
- Polymerase Chain Reaction * methods MeSH
- Antibodies, Bacterial blood MeSH
- Sensitivity and Specificity MeSH
- Syphilis Serodiagnosis methods MeSH
- Syphilis * diagnosis microbiology MeSH
- Treponema pallidum * genetics isolation & purification immunology MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Young Adult MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
BACKGROUND: The increase in syphilis rates worldwide necessitates development of a vaccine with global efficacy. We aimed to explore Treponema pallidum subspecies pallidum (TPA) molecular epidemiology essential for vaccine research by analysing clinical data and specimens from early syphilis patients using whole-genome sequencing (WGS) and publicly available WGS data. METHODS: In this multicentre, cross-sectional, molecular epidemiology study, we enrolled patients with primary, secondary, or early latent syphilis from clinics in China, Colombia, Malawi, and the USA between Nov 28, 2019, and May 27, 2022. Participants aged 18 years or older with laboratory confirmation of syphilis by direct detection methods or serological testing, or both, were included. Patients were excluded from enrolment if they were unwilling or unable to give informed consent, did not understand the study purpose or nature of their participation, or received antibiotics active against syphilis in the past 30 days. TPA detection and WGS were conducted on lesion swabs, skin biopsies, skin scrapings, whole blood, or rabbit-passaged isolates. We compared our WGS data to publicly available genomes and analysed TPA populations to identify mutations associated with lineage and geography. FINDINGS: We screened 2802 patients and enrolled 233 participants, of whom 77 (33%) had primary syphilis, 154 (66%) had secondary syphilis, and two (1%) had early latent syphilis. The median age of participants was 28 years (IQR 22-35); 154 (66%) participants were cisgender men, 77 (33%) were cisgender women, and two (1%) were transgender women. Of the cisgender men, 66 (43%) identified as gay, bisexual, or other sexuality. Among all participants, 56 (24%) had HIV co-infection. WGS data from 113 participants showed a predominance of SS14-lineage strains with geographical clustering. Phylogenomic analyses confirmed that Nichols-lineage strains were more genetically diverse than SS14-lineage strains and clustered into more distinct subclades. Differences in single nucleotide variants (SNVs) were evident by TPA lineage and geography. Mapping of highly differentiated SNVs to three-dimensional protein models showed population-specific substitutions, some in outer membrane proteins (OMPs) of interest. INTERPRETATION: Our study substantiates the global diversity of TPA strains. Additional analyses to explore TPA OMP variability within strains is vital for vaccine development and understanding syphilis pathogenesis on a population level. FUNDING: US National Institutes of Health National Institute for Allergy and Infectious Disease, the Bill & Melinda Gates Foundation, Connecticut Children's, and the Czech Republic National Institute of Virology and Bacteriology.
- MeSH
- Bacterial Vaccines immunology administration & dosage MeSH
- Adult MeSH
- Phylogeny MeSH
- Genetic Variation genetics MeSH
- Genome, Bacterial MeSH
- Genomics MeSH
- Middle Aged MeSH
- Humans MeSH
- Young Adult MeSH
- Molecular Epidemiology * MeSH
- Cross-Sectional Studies MeSH
- Whole Genome Sequencing * MeSH
- Syphilis * epidemiology microbiology MeSH
- Treponema pallidum * genetics immunology MeSH
- Treponema MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Young Adult MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Multicenter Study MeSH
- Geographicals
- United States MeSH
The treponemes infecting lagomorphs include Treponema paraluisleporidarum ecovar Cuniculus (TPeC) and ecovar Lepus (TPeL), infecting rabbits and hares, respectively. In this study, we described the first complete genome sequence of TPeL, isolate V3603-13, from an infected mountain hare (Lepus timidus) in Sweden. In addition, we determined 99.0% of the genome sequence of isolate V246-08 (also from an infected mountain hare, Sweden) and 31.7% of the genome sequence of isolate Z27 A77/78 (from a European hare, Lepus europeaus, The Netherlands). The TPeL V3603-13 genome had considerable gene synteny with the TPeC Cuniculi A genome and with the human pathogen T. pallidum, which causes syphilis (ssp. pallidum, TPA), yaws (ssp. pertenue, TPE) and endemic syphilis (ssp. endemicum, TEN). Compared to the TPeC Cuniculi A genome, TPeL V3603-13 contained four insertions and 11 deletions longer than three nucleotides (ranging between 6 and2,932 nts). In addition, there were 25 additional indels, from one to three nucleotides long, altogether spanning 36 nts. The number of single nucleotide variants (SNVs) between TPeC Cuniculi A and TPeL V3603-13 were represented by 309 nucleotide differences. Major proteome coding differences between TPeL and TPeC were found in the tpr gene family, and (predicted) genes coding for outer membrane proteins, suggesting that these components are essential for host adaptation in lagomorph syphilis. The phylogeny revealed that the TPeL sample from the European brown hare was more distantly related to TPeC Cuniculi A than V3603-13 and V246-08.
- MeSH
- Phylogeny * MeSH
- Genome, Bacterial MeSH
- Rabbits MeSH
- Syphilis * microbiology MeSH
- Treponema * genetics isolation & purification MeSH
- Hares * microbiology MeSH
- Animals MeSH
- Check Tag
- Rabbits MeSH
- Animals MeSH
- Publication type
- Journal Article 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
- Alleles * MeSH
- Adult MeSH
- Genotype MeSH
- Middle Aged MeSH
- Humans MeSH
- Macrolides pharmacology MeSH
- Multilocus Sequence Typing * MeSH
- Syphilis * microbiology epidemiology genetics MeSH
- Treponema pallidum * genetics isolation & purification MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
The etiological agent of yaws is the spirochete Treponema pallidum (TP) subsp. pertenue (TPE) and infects the children of Papua New Guinea, causing ulcerative skin lesions that impairs normal growth and development. Closely related strains of Treponema pallidum subsp. pertenue, JE11, and TE13 were detected in an ulcer biospecimen derived from a 5-year-old yaws patient. Cloning experiments validated the presence of two distinct but similar genotypes, namely TE13 and JE11, co-occurring within a single host. While coinfection with highly related TPE strains has only limited epidemiological and clinical relevance, this is the first documented coinfection with genetically distinct TP strains in a single patient. Similar coinfections in the past were explained by the existence of over a dozen recombinant loci present in the TP genomes as a result of inter-strain or inter-subspecies recombination events following an anticipated scenario of TP coinfection, i.e., uptake of foreign DNA and DNA recombination.
- MeSH
- DNA, Bacterial genetics MeSH
- Yaws * microbiology MeSH
- Phylogeny MeSH
- Genotype * MeSH
- Coinfection * microbiology MeSH
- Humans MeSH
- Child, Preschool MeSH
- Sequence Analysis, DNA MeSH
- Treponema pallidum * genetics isolation & purification classification MeSH
- Treponema MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Child, Preschool MeSH
- Publication type
- Journal Article MeSH
- Case Reports MeSH
- Geographicals
- Papua New Guinea MeSH
Yaws is an endemic disease caused by Treponema pallidum subsp. pertenue (TPE) that primarily affects children in rural regions of the tropics. The endemic character of yaws infections and the expected exclusive reservoir of TPE in humans opened a new opportunity to start a yaws eradication campaign. We have developed a multi-locus sequence typing (MLST) scheme for TPE isolates combining the previously published (TP0548, TP0488) and new (TP0858) chromosomal loci, and we compared this typing scheme to the two previously published MLST schemes. We applied this scheme to TPE-containing clinical isolates obtained during a mass drug administration study performed in the Namatanai District of Papua New Guinea between June 2018 and December 2019. Of 1081 samples collected, 302 (28.5%) tested positive for TPE DNA, from which 255 (84.4%) were fully typed. The TPE PCR-positivity in swab samples was higher in younger patients, patients with single ulcers, first ulcer episodes, and with ulcer duration less than six months. Non-treponemal serological test positivity correlated better with PCR positivity compared to treponema-specific serological tests. The MLST revealed a low level of genetic diversity among infecting TPE isolates, represented by just three distinct genotypes (JE11, SE22, and TE13). Two previously used typing schemes revealed similar typing resolutions. Two new alleles (one in TP0858 and one in TP0136) were shown to arise by intragenomic recombination/deletion events. Compared to samples genotyped as JE11, the minor genotypes (TE13 and SE22) were more frequently detected in samples from patients with two or more ulcers and patients with higher values of specific TP serological tests. Moreover, the A2058G mutation in the 23S rRNA genes of three JE11 isolates was found, resulting in azithromycin resistance.
