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Michal Strouhal - One of the best experts on this subject based on the ideXlab platform.
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nonhuman primates across sub saharan africa are infected with the yaws bacterium Treponema pallidum subsp pertenue
Emerging microbes & infections, 2018Co-Authors: Sascha Knauf, Michal Strouhal, Idrissa S Chuma, Lenka Mikalova, Roy Armstrong, Jan F Gogarten, Verena J Schuenemann, E K Batamuzi, Bernard Davoust, Georges DiattaAbstract:The bacterium Treponema pallidum (TP) causes human syphilis (subsp. pallidum; TPA), bejel (subsp. endemicum; TEN), and yaws (subsp. pertenue; TPE). Although syphilis has reached a worldwide distribution, bejel and yaws have remained endemic diseases. Bejel affects individuals in dry areas of Sahelian Africa and Saudi Arabia, whereas yaws affects those living in the humid tropics. Yaws is currently reported as endemic in 14 countries, and an additional 84 countries have a known history of yaws but lack recent epidemiological data. Although this disease was subject to global eradication efforts in the mid-20th century, it later reemerged in West Africa, Southern Asia, and the Pacific region5. New large-scale treatment options triggered the ongoing second eradication campaign, the goal of which is to eradicate yaws globally by 2020.
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sequencing of Treponema pallidum subsp pallidum from isolate uz1974 using anti Treponemal antibodies enrichment first complete whole genome sequence obtained directly from human clinical material
PLOS ONE, 2018Co-Authors: Linda Grillova, Christina M Marra, Arturo Centurionlara, Lorenzo Giacani, Michal Strouhal, Lenka Mikalova, Radim Strnadel, Lucy Poveda, Giancarlo Russo, Darina CejkovaAbstract:Treponema pallidum subsp. pallidum (TPA) is the infectious agent of syphilis, a disease that infects more than 5 million people annually. Since TPA is an uncultivable bacterium, most of the information on TPA genetics comes from genome sequencing and molecular typing studies. This study presents the first complete TPA genome (without sequencing gaps) of clinical isolate (UZ1974), which was obtained directly from clinical material, without multiplication in rabbits. Whole genome sequencing was performed using a newly developed Anti-Treponemal Antibody Enrichment technique combined with previously reported Pooled Segment Genome Sequencing. We identified the UW074B genome, isolated from a sample previously propagated in rabbits, to be the closest relative of the UZ1974 genome and calculated the TPA mutation rate as 2.8 x 10(-10) per site per generation.
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molecular characterization of Treponema pallidum subsp pallidum in switzerland and france with a new multilocus sequence typing scheme
PLOS ONE, 2018Co-Authors: Linda Grillova, Michal Strouhal, Angele Gayetagero, P Sednaoui, Trista Ferry, Matthias Cavassini, Lenka Mikalova, Tanika Awa, Kay Niesel, Stepha LautenschlageAbstract:Syphilis is an important public health problem and an increasing incidence has been noted in recent years. Characterization of strain diversity through molecular data plays a critical role in the epidemiological understanding of this re-emergence. We here propose a new high-resolution multilocus sequence typing (MLST) scheme for Treponema pallidum subsp. pallidum (TPA). We analyzed 30 complete and draft TPA genomes obtained directly from clinical samples or from rabbit propagated strains to identify suitable typing loci and tested the new scheme on 120 clinical samples collected in Switzerland and France. Our analyses yielded three loci with high discriminatory power: TP0136, TP0548, and TP0705. Together with analysis of the 23S rRNA gene mutations for macrolide resistance, we propose these loci as MLST for TPA. Among clinical samples, 23 allelic profiles as well as a high percentage (80% samples) of macrolide resistance were revealed. The new MLST has higher discriminatory power compared to previous typing schemes, enabling distinction of TPA from other Treponemal bacteria, distinction between the two main TPA clades (Nichols and SS14), and differentiation of strains within these clades.
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african nonhuman primates are infected with the yaws bacterium Treponema pallidum subsp pertenue
bioRxiv, 2017Co-Authors: Sascha Knauf, Ja F Gogarte, Verena J Schuenema, Michal Strouhal, E K Atamuzi, Helene M De Nys, Ariane Due, Lenka Mikalova, Roy Armstrong, Idrissa S ChumaAbstract:Treponema pallidum subsp. pertenue (TPE) is the causative agent of yaws. The disease was subject to global eradication efforts in the mid 20th century but reemerged in West Africa, Southern Asia, and the Pacific region. Despite its importance for eradication, detailed data on possible nonhuman disease reservoirs are missing. A number of African nonhuman primates (NHPs) have been reported to show skin ulcerations suggestive of Treponemal infection in humans. Furthermore antibodies against Treponema pallidum (TP) have been repeatedly detected in wild NHP populations. While genetic studies confirmed that NHPs are infected with TP strains, subspecies identification was only possible once for a strain isolated in 1966, pinpointing the involvement of TPE. We therefore collected a number of recently isolated simian TP strains and determined eight whole genome sequences using hybridization capture or long-range PCR combined with next-generation sequencing. These new genomes were compared with those of known human TP isolates. Our results show that naturally occurring simian TP strains circulating in three African NHP species all cluster with human TPE strains and show the same genomic structure as human TPE strains. These data indicate that humans are not the exclusive host for the yaws bacterium and that a One Health approach is required to achieve sustainable eradication of human yaws.
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origin of modern syphilis and emergence of a pandemic Treponema pallidum cluster
Nature microbiology, 2017Co-Authors: Natasha Arora, Verena J Schuenema, Michal Strouhal, Alexande Herbig, Gunte Jage, Alexande Peltze, Alexande Seitz, Linda Grillova, Leono SanchezbusoAbstract:The abrupt onslaught of the syphilis pandemic that started in the late fifteenth century established this devastating infectious disease as one of the most feared in human history 1. Surprisingly, despite the availability of effective antibiotic treatment since the mid-twentieth century, this bacterial infection, which is caused by Treponema pallidum subsp. pallidum (TPA), has been re-emerging globally in the last few decades with an estimated 10.6 million cases in 2008 (ref. 2). Although resistance to penicillin has not yet been identified, an increasing number of strains fail to respond to the second-line antibiotic azithromycin 3. Little is known about the genetic patterns in current infections or the evolutionary origins of the disease due to the low quantities of Treponemal DNA in clinical samples and difficulties in cultivating the pathogen 4. Here, we used DNA capture and whole-genome sequencing to successfully interrogate genome-wide variation from syphilis patient specimens, combined with laboratory samples of TPA and two other subspecies. Phylogenetic comparisons based on the sequenced genomes indicate that the TPA strains examined share a common ancestor after the fifteenth century, within the early modern era. Moreover, most contemporary strains are azithromycin-resistant and are members of a globally dominant cluster, named here as SS14-omega. The cluster diversified from a common ancestor in the mid-twentieth century subsequent to the discovery of antibiotics. Its recent phylogenetic divergence and global presence point to the emergence of a pandemic strain cluster.
Darina Cejkova - One of the best experts on this subject based on the ideXlab platform.
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sequencing of Treponema pallidum subsp pallidum from isolate uz1974 using anti Treponemal antibodies enrichment first complete whole genome sequence obtained directly from human clinical material
PLOS ONE, 2018Co-Authors: Linda Grillova, Christina M Marra, Arturo Centurionlara, Lorenzo Giacani, Michal Strouhal, Lenka Mikalova, Radim Strnadel, Lucy Poveda, Giancarlo Russo, Darina CejkovaAbstract:Treponema pallidum subsp. pallidum (TPA) is the infectious agent of syphilis, a disease that infects more than 5 million people annually. Since TPA is an uncultivable bacterium, most of the information on TPA genetics comes from genome sequencing and molecular typing studies. This study presents the first complete TPA genome (without sequencing gaps) of clinical isolate (UZ1974), which was obtained directly from clinical material, without multiplication in rabbits. Whole genome sequencing was performed using a newly developed Anti-Treponemal Antibody Enrichment technique combined with previously reported Pooled Segment Genome Sequencing. We identified the UW074B genome, isolated from a sample previously propagated in rabbits, to be the closest relative of the UZ1974 genome and calculated the TPA mutation rate as 2.8 x 10(-10) per site per generation.
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resequencing of Treponema pallidum ssp pallidum strains nichols and ss14 correction of sequencing errors resulted in increased separation of syphilis treponeme subclusters
PLOS ONE, 2013Co-Authors: Helena Pětrosova, Michal Strouhal, Marie Zobaníková, Petra Pospíšilová, Lenka Mikalova, Darina CejkovaAbstract:Article on the resequencing of Treponema pallidum ssp. pallidum strains nichols and SS14 and how the correction of sequencing errors resulted in increased separation of syphilis treponeme subclusters.
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p1 011 whole genome sequence of the Treponema pallidum ssp endemicum strain bosnia a
Sexually Transmitted Infections, 2013Co-Authors: Arbora Staudova, Arturo Centurionlara, Lorenzo Giacani, Michal Strouhal, Marie Zobaníková, George M. Weinstock, Darina Cejkova, Lei Che, Lucinda Fulto, David SmajsAbstract:Background Treponema pallidum ssp. endemicum (TEN) is the causative agent of endemic syphilis (bejel). The TEN Bosnia A strain was isolated in 1950 from a patient’s penile lesion in northeastern Bosnia. Methods To define genetic differences between TEN Bosnia A and other pathogenic treponemes including the agents of syphilis ( T. pallidum ssp. pallidum , TPA)and yaws ( T. pallidum ssp. pertenue , TPE), a high quality sequence of the Bosnia A genome was determined using 454-pyrosequencing, Illumina, SOLiD and traditional Sanger sequencing. Combined average coverage of these sequencing methods was greater than 340x. Results Compared to other TPA and TPE treponemes, the genome of Bosnia A (1,137,653 bp) was smaller in size (∼2 kb) but structurally almost identical to other TPA and TPE strains. The Bosnia A genome clustered with TPE strains (nucleotide identity excluding indels ranged between 99.91 – 99.94%) while TPAstrains were more distantly related (99.79 – 99.82%). More than 400 Bosnia A-specific nt changes (i.e. sequences different from TPA and TEN genomes) were found as the result of our analysis. Conclusions The Bosnia A genome showed similar genetic characteristics as other TPA and TPE strains. Genetic differences found between TPA strains and Bosnia A genome could be used for identification of potential virulence factors of syphis treponemes. Moreover, genetic changes specific for Bosnia A genome could help develop molecular diagnostic tests for endemic syphilis.
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whole genome sequence of Treponema pallidum ssp pallidum strain mexico a suggests recombination between yaws and syphilis strains
PLOS Neglected Tropical Diseases, 2012Co-Authors: Helena Pětrosova, Michal Strouhal, Marie Zobaníková, Petra Pospíšilová, Lenka Mikalova, Darina Cejkova, Lei CheAbstract:Treponema pallidum is a Gram-negative spirochete that causes diseases with distinct clinical manifestations and uses different transmission strategies. While syphilis (caused by subspecies pallidum) is a worldwide venereal and congenital disease, yaws (caused by subspecies pertenue) is a tropical disease transmitted by direct skin contact. Currently the genetic basis and evolution of these diseases remain unknown. In this study, we describe a high quality whole genome sequence of T. pallidum ssp. pallidum strain Mexico A, determined using the ?next generation? sequencing technique (Illumina). Although the genome of this strain contains no large rearrangements in comparison with other Treponemal genomes, we found two genes which combined sequences from both subspecies pallidum and pertenue. The observed mosaic character of these two genes is likely a result of inter-strain recombination between pallidum and pertenue during simultaneous infection of a single host.
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whole genome sequences of three Treponema pallidum ssp pertenue strains yaws and syphilis treponemes differ in less than 0 2 of the genome sequence
PLOS Neglected Tropical Diseases, 2012Co-Authors: Michal Strouhal, Marie Zobaníková, Petra Pospíšilová, Darina Cejkova, Lei CheAbstract:Spirochete Treponema pallidum ssp. pertenue (TPE) is the causative agent of yaws while strains of Treponema pallidum ssp. pallidum (TPA) cause syphilis. Both yaws and syphilis are distinguished on the basis of epidemiological characteristics and clinical symptoms. Neither treponeme can reproduce outside the host organism, which precludes the use of standard molecular biology techniques used to study cultivable pathogens. In this study, we determined high quality whole genome sequences of TPE strains and compared them to known genetic information for T. pallidum ssp. pallidum strains. The genome structure was identical in all three TPE strains and also between TPA and TPE strains. The TPE genome length ranged between 1,139,330 bp and 1,139,744 bp. The overall sequence identity between TPA and TPE genomes was 99.8%, indicating that the two pathogens are extremely closely related. A set of 34 TPE genes (3.5%) encoded proteins containing six or more amino acid replacements or other major sequence changes. These genes more often belonged to the group of genes with predicted virulence and unknown functions suggesting their involvement in infection differences between yaws and syphilis.
Christina M Marra - One of the best experts on this subject based on the ideXlab platform.
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comparative genomics and full length tprk profiling of Treponema pallidum subsp pallidum reinfection
PLOS Neglected Tropical Diseases, 2020Co-Authors: Ami Addetia, Laure C Tantalo, Christina M Marra, Hong Xie, Meeili Huang, Alexande L GreningeAbstract:Developing a vaccine against Treponema pallidum subspecies pallidum, the causative agent of syphilis, remains a public health priority. Syphilis vaccine design efforts have been complicated by lack of an in vitro T. pallidum culture system, prolific antigenic variation in outer membrane protein TprK, and lack of functional annotation for nearly half of the genes. Understanding the genetic basis of T. pallidum reinfection can provide insights into variation among strains that escape cross-protective immunity. Here, we present comparative genomic sequencing and deep, full-length tprK profiling of two T. pallidum isolates from blood from the same patient that were collected six years apart. Notably, this patient was diagnosed with syphilis four times, with two of these episodes meeting the definition of neurosyphilis, during this interval. Outside of the highly variable tprK gene, we identified 14 coding changes in 13 genes. Nine of these genes putatively localized to the periplasmic or outer membrane spaces, consistent with a potential role in serological immunoevasion. Using a newly developed full-length tprK deep sequencing protocol, we profiled the diversity of this gene that far outpaces the rest of the genome. Intriguingly, we found that the reinfecting isolate demonstrated less diversity across each tprK variable region compared to the isolate from the first infection. Notably, the two isolates did not share any full-length TprK sequences. Our results are consistent with an immunodominant-evasion model in which the diversity of TprK explains the ability of T. pallidum to successfully reinfect individuals, even when they have been infected with the organism multiple times.
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comparative genomics and full length tprk profiling of Treponema pallidum subsp pallidum reinfection
bioRxiv, 2019Co-Authors: Ami Addetia, Laure C Tantalo, Christina M Marra, Hong Xie, Meeili Huang, Alexande L GreningeAbstract:Abstract Developing a vaccine against Treponema pallidum subspecies pallidum, the causative agent of syphilis, remains a public health priority. Syphilis vaccine design efforts have been complicated by lack of an in vitro T. pallidum culture system, prolific antigenic variation in outer membrane protein TprK, and lack of functional annotation for nearly half of the genes. Understanding the genetic basis of T. pallidum reinfection can provide insights into variation among strains that escape cross-protective immunity. Here, we present comparative genomic sequencing and deep, full-length tprK profiling of two T. pallidum isolates from blood from the same patient that were collected six years apart. Notably, this patient was diagnosed with syphilis four times, with two of these episodes meeting the definition of neurosyphilis, during this interval. Outside of the highly variable tprK gene, we identified 14 coding changes in 13 genes. Nine of these genes putatively localized to the periplasmic or outer membrane spaces, consistent with a potential role in serological immunoevasion. Using a newly developed full-length tprK deep sequencing protocol, we profiled the diversity of this gene that far outpaces the rest of the genome. Intriguingly, we found that the reinfecting isolate demonstrated less diversity across each tprK variable region compared to the isolate from the first infection. Notably, the two isolates did not share any full-length TprK sequences. Our results are consistent with an immunodominant-evasion model in which the diversity of TprK explains the ability of T. pallidum to successfully reinfect individuals, even when they have been infected with the organism multiple times. Author Summary The causative agent of syphilis, Treponema pallidum subspecies pallidum, is capable of repeat infections in people, suggesting that the human immune response does not develop sufficiently broad or long-lasting immunity to cover Treponemal diversity. Here, we examined the genomes from two blood-derived isolates of T. pallidum derived 6 years apart from a patient who had syphilis four times during the same period to understand the genetic basis of reinfection. We found a paucity of coding changes across the genome outside of the highly variable tprK gene. Using deep profiling of the full-length tprK gene, we found surprisingly that the two isolates did not share any full-length TprK sequences.
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p750 a comparison of Treponema pallidum molecular typing systems mlst vs ecdct
Sexually Transmitted Infections, 2019Co-Authors: Sharo K Sahi, Laure C Tantalo, Christina M MarraAbstract:Background Several syphilis typing systems have been proposed. Recent work suggests that multilocus sequence typing (MLST) may be superior to enhanced CDC typing (ECDCT), particularly because ECDCT type may differ among organisms amplified from different anatomical sites in the same person. The goal of this study was to compare the two systems. Methods DNA was extracted from 20 Treponema pallidum isolates propagated in rabbits, 10 oral and 10 genital or non-genital lesion swabs, and 10 blood samples from patients with syphilis. MLST type for tp0136, tp0548 and tp0705 and ECDCT type were determined according to published methods. Samples were chosen because they were completely typeable by ECDCT. ECDCT types were also determined for samples from different anatomical sites in 7 patients, and from blood and blood isolates (rabbit propagated) in 8 patients. Results MLST type could be fully determined for 19 (95%) of 20 bacterial isolates, 8 (80%) of 10 bloods, 7 (70%) of 10 lesion swabs, and 5 (50%) of 10 oral swabs. 13 subtypes were identified by ECDCT, and 12 by MLST. While MLST was able to subdivide two common ECDCT types (1.1.1, 1.1.2, 1.1.9, and 1.37.1 within 14d/f; and 1.3.1, 1.38.1, and 6.3.1 within 14d/g), it failed to distinguish among less common ECDCT types. ECDCT type was identical in 5 paired lesion and oral swabs, 1 paired blood and oral swab, and 1 paired blood, lesion and oral swabs. In addition, ECDCT type was identical in 8 paired blood and blood isolates. Conclusion Compared to ECDCT, determination of MLST was less often successful from isolates and from clinical samples, and it was not uniformly more discriminating. ECDCT was stable among anatomical sites and between direct patient-derived samples compared to rabbit propagated organisms. Disclosure No significant relationships.
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p762 is kissing safe detection of Treponema pallidum in oral swabs from patients with syphilis
Sexually Transmitted Infections, 2019Co-Authors: Laure C Tantalo, Sharo K Sahi, David A Katz, Christina M MarraAbstract:Background Treponema pallidum (TP) DNA has been detected in oral swabs from patients with early syphilis. The goal of this study was to determine the frequency of detectable TP DNA in oral swabs from patients with all stages of syphilis, and from appropriate controls. Methods Blood (N = 131), oral swabs (N = 112) and lesion swabs (N = 72) were collected from 138 patients with untreated syphilis (cases). Controls were oral swabs collected from 89 patients presenting to an STD clinic for a concern other than syphilis, 59 HIV positive individuals, and 108 individuals 3, 6 or 12 months after treatment of uncomplicated or neurosyphilis (194 samples). DNA was extracted and underwent amplification of a portion of the TP tp0548 gene. Rapid plasma reagin (RPR) tests were performed on serum, and the association between RPR titer and oral TP detection assessed using Mann-Whitney U test. Results Among those with known syphilis stage, TP was detected in oral swabs from 31/101 (31%) cases: 4/29 (14%) with primary, 20/49 (41%) with secondary, 6/15 (40%) with early latent, 1/7 (14%) with late latent syphilis, and 0/1 (0%) syphilis contact. Oral TP was detected in 18/74 (24%) without detectable blood TP, 6/18 (33%) without detectable lesion TP, and 21/78 (27%) without oral lesions. Oral TP detection was more likely with higher RPR titers (p = 0.002). Oral TP was not detected in any of the 342 control samples. Conclusion Oral TP is detectable in patients at all syphilis stages, most commonly when serum RPR titer is high. Oral TP can be detected even when undetectable in blood or lesion swabs, and in the absence of oral lesions. While detection of DNA is not the same as detection of virulent organisms, these results suggest that individuals with syphilis could infect their partners solely through oral contact. Disclosure No significant relationships.
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sequencing of Treponema pallidum subsp pallidum from isolate uz1974 using anti Treponemal antibodies enrichment first complete whole genome sequence obtained directly from human clinical material
PLOS ONE, 2018Co-Authors: Linda Grillova, Christina M Marra, Arturo Centurionlara, Lorenzo Giacani, Michal Strouhal, Lenka Mikalova, Radim Strnadel, Lucy Poveda, Giancarlo Russo, Darina CejkovaAbstract:Treponema pallidum subsp. pallidum (TPA) is the infectious agent of syphilis, a disease that infects more than 5 million people annually. Since TPA is an uncultivable bacterium, most of the information on TPA genetics comes from genome sequencing and molecular typing studies. This study presents the first complete TPA genome (without sequencing gaps) of clinical isolate (UZ1974), which was obtained directly from clinical material, without multiplication in rabbits. Whole genome sequencing was performed using a newly developed Anti-Treponemal Antibody Enrichment technique combined with previously reported Pooled Segment Genome Sequencing. We identified the UW074B genome, isolated from a sample previously propagated in rabbits, to be the closest relative of the UZ1974 genome and calculated the TPA mutation rate as 2.8 x 10(-10) per site per generation.
Lenka Mikalova - One of the best experts on this subject based on the ideXlab platform.
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nonhuman primates across sub saharan africa are infected with the yaws bacterium Treponema pallidum subsp pertenue
Emerging microbes & infections, 2018Co-Authors: Sascha Knauf, Michal Strouhal, Idrissa S Chuma, Lenka Mikalova, Roy Armstrong, Jan F Gogarten, Verena J Schuenemann, E K Batamuzi, Bernard Davoust, Georges DiattaAbstract:The bacterium Treponema pallidum (TP) causes human syphilis (subsp. pallidum; TPA), bejel (subsp. endemicum; TEN), and yaws (subsp. pertenue; TPE). Although syphilis has reached a worldwide distribution, bejel and yaws have remained endemic diseases. Bejel affects individuals in dry areas of Sahelian Africa and Saudi Arabia, whereas yaws affects those living in the humid tropics. Yaws is currently reported as endemic in 14 countries, and an additional 84 countries have a known history of yaws but lack recent epidemiological data. Although this disease was subject to global eradication efforts in the mid-20th century, it later reemerged in West Africa, Southern Asia, and the Pacific region5. New large-scale treatment options triggered the ongoing second eradication campaign, the goal of which is to eradicate yaws globally by 2020.
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sequencing of Treponema pallidum subsp pallidum from isolate uz1974 using anti Treponemal antibodies enrichment first complete whole genome sequence obtained directly from human clinical material
PLOS ONE, 2018Co-Authors: Linda Grillova, Christina M Marra, Arturo Centurionlara, Lorenzo Giacani, Michal Strouhal, Lenka Mikalova, Radim Strnadel, Lucy Poveda, Giancarlo Russo, Darina CejkovaAbstract:Treponema pallidum subsp. pallidum (TPA) is the infectious agent of syphilis, a disease that infects more than 5 million people annually. Since TPA is an uncultivable bacterium, most of the information on TPA genetics comes from genome sequencing and molecular typing studies. This study presents the first complete TPA genome (without sequencing gaps) of clinical isolate (UZ1974), which was obtained directly from clinical material, without multiplication in rabbits. Whole genome sequencing was performed using a newly developed Anti-Treponemal Antibody Enrichment technique combined with previously reported Pooled Segment Genome Sequencing. We identified the UW074B genome, isolated from a sample previously propagated in rabbits, to be the closest relative of the UZ1974 genome and calculated the TPA mutation rate as 2.8 x 10(-10) per site per generation.
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molecular characterization of Treponema pallidum subsp pallidum in switzerland and france with a new multilocus sequence typing scheme
PLOS ONE, 2018Co-Authors: Linda Grillova, Michal Strouhal, Angele Gayetagero, P Sednaoui, Trista Ferry, Matthias Cavassini, Lenka Mikalova, Tanika Awa, Kay Niesel, Stepha LautenschlageAbstract:Syphilis is an important public health problem and an increasing incidence has been noted in recent years. Characterization of strain diversity through molecular data plays a critical role in the epidemiological understanding of this re-emergence. We here propose a new high-resolution multilocus sequence typing (MLST) scheme for Treponema pallidum subsp. pallidum (TPA). We analyzed 30 complete and draft TPA genomes obtained directly from clinical samples or from rabbit propagated strains to identify suitable typing loci and tested the new scheme on 120 clinical samples collected in Switzerland and France. Our analyses yielded three loci with high discriminatory power: TP0136, TP0548, and TP0705. Together with analysis of the 23S rRNA gene mutations for macrolide resistance, we propose these loci as MLST for TPA. Among clinical samples, 23 allelic profiles as well as a high percentage (80% samples) of macrolide resistance were revealed. The new MLST has higher discriminatory power compared to previous typing schemes, enabling distinction of TPA from other Treponemal bacteria, distinction between the two main TPA clades (Nichols and SS14), and differentiation of strains within these clades.
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african nonhuman primates are infected with the yaws bacterium Treponema pallidum subsp pertenue
bioRxiv, 2017Co-Authors: Sascha Knauf, Ja F Gogarte, Verena J Schuenema, Michal Strouhal, E K Atamuzi, Helene M De Nys, Ariane Due, Lenka Mikalova, Roy Armstrong, Idrissa S ChumaAbstract:Treponema pallidum subsp. pertenue (TPE) is the causative agent of yaws. The disease was subject to global eradication efforts in the mid 20th century but reemerged in West Africa, Southern Asia, and the Pacific region. Despite its importance for eradication, detailed data on possible nonhuman disease reservoirs are missing. A number of African nonhuman primates (NHPs) have been reported to show skin ulcerations suggestive of Treponemal infection in humans. Furthermore antibodies against Treponema pallidum (TP) have been repeatedly detected in wild NHP populations. While genetic studies confirmed that NHPs are infected with TP strains, subspecies identification was only possible once for a strain isolated in 1966, pinpointing the involvement of TPE. We therefore collected a number of recently isolated simian TP strains and determined eight whole genome sequences using hybridization capture or long-range PCR combined with next-generation sequencing. These new genomes were compared with those of known human TP isolates. Our results show that naturally occurring simian TP strains circulating in three African NHP species all cluster with human TPE strains and show the same genomic structure as human TPE strains. These data indicate that humans are not the exclusive host for the yaws bacterium and that a One Health approach is required to achieve sustainable eradication of human yaws.
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molecular typing of Treponema pallidum isolates from buenos aires argentina frequent nichols like isolates and low levels of macrolide resistance
PLOS ONE, 2017Co-Authors: Lucia Gallo Vaule, Lenka Mikalova, Linda Grillova, Ricardo Casco, Marcelo Rodriguez Fermepi, Maria A Pando, David SmajsAbstract:A total of 54 clinical samples, including genital lesion swabs, whole blood and cerebrospinal fluid from patients diagnosed with syphilis were collected in 2006 and in 2013 in Buenos Aires, Argentina. Treponemal DNA was detected in 43 of the analyzed samples (79.6%) and further analyzed using Sequencing-based molecular typing (SBMT) and Enhanced CDC-typing (ECDCT). By SBMT, 10 different Treponema pallidum subsp. pallidum (TPA) genotypes were found, of which six were related to the TPA SS14 strain, and four to the TPA Nichols strain. The 23S rRNA gene was amplified in samples isolated from 42 patients, and in six of them (14.3%), either the A2058G (four patients, 9.5%) or the A2059G (two patients, 4.8%) mutations were found. In addition to Taiwan, Madagascar and Peru, Argentina is another country where the prevalence of Nichols-like isolates (26.8%) is greater than 10%.
Sheila A Lukeha - One of the best experts on this subject based on the ideXlab platform.
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Treponema pallidum nucleic acid amplification testing to augment syphilis screening among men who have sex with men
Journal of Clinical Microbiology, 2019Co-Authors: Charmie Godornes, Sheila A Lukeha, Matthew R Golde, Megha Odonnell, Paul D Swenso, Paul Hovey, Sarah S Romano, Damo K GetmaAbstract:Syphilis rates in much of the world are now at their highest levels in almost three decades, and new approaches to controlling syphilis, including diagnostic tests with shorter window periods, are urgently needed. We compared the sensitivity of syphilis serological testing using the rapid plasma reagin (RPR) test with that of the combination of serological testing and an experimental 23S rRNA Treponema pallidum real-time transcription-mediated amplification (TMA) assay performed on rectal and pharyngeal mucosal swabs. T. pallidum PCR assays for the tpp47 gene were performed on all TMA-positive specimens, as well as specimens from 20 randomly selected TMA-negative men. A total of 545 men who have sex with men (MSM) who were seen in a sexually transmitted disease clinic provided 506 pharyngeal specimens and 410 rectal specimens with valid TMA results. Twenty-two men (4%) were diagnosed with syphilis on the basis of positive RPR test results and clinical diagnoses, including 3 men with primary infections, 8 with secondary syphilis, 9 with early latent syphilis, 1 with late latent syphilis, and 1 with an unstaged infection. Two additional men were diagnosed based on positive rectal mucosal TMA assay results alone, and both also tested positive by PCR assay. At least 1 specimen was TMA positive for 12 of 24 men with syphilis (sensitivity, 50% [95% confidence interval [CI], 29 to 71%]). RPR testing and clinical diagnosis were 92% sensitive (95% CI, 73 to 99%) in identifying infected men. Combining mucosal TMA testing and serological testing may increase the sensitivity of syphilis screening in high-risk populations.
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diagnostics for yaws eradication insights from direct next generation sequencing of cutaneous strains of Treponema pallidum
Clinical Infectious Diseases, 2018Co-Authors: Michael Marks, Sheila A Lukeha, Maria Fookes, Josef Wagne, Robe Utche, Rosanna Ghinai, Olive Sokana, Yawadu Sarkodie, Anthony W Solomo, David MabeyAbstract:Background: Yaws-like chronic ulcers can be caused by Treponema pallidum subspecies pertenue, Haemophilus ducreyi, or other, still-undefined bacteria. To permit accurate evaluation of yaws elimination efforts, programmatic use of molecular diagnostics is required. The accuracy and sensitivity of current tools remain unclear because our understanding of T. pallidum diversity is limited by the low number of sequenced genomes. Methods: We tested samples from patients with suspected yaws collected in the Solomon Islands and Ghana. All samples were from patients whose lesions had previously tested negative using the Centers for Disease Control and Prevention (CDC) diagnostic assay in widespread use. However, some of these patients had positive serological assays for yaws on blood. We used direct whole-genome sequencing to identify T. pallidum subsp pertenue strains missed by the current assay. Results: From 45 Solomon Islands and 27 Ghanaian samples, 11 were positive for T. pallidum DNA using the species-wide quantitative polymerase chain reaction (PCR) assay, from which we obtained 6 previously undetected T. pallidum subsp pertenue whole-genome sequences. These show that Solomon Islands sequences represent distinct T. pallidum subsp pertenue clades. These isolates were invisible to the CDC diagnostic PCR assay, due to sequence variation in the primer binding site. Conclusions: Our data double the number of published T. pallidum subsp pertenue genomes. We show that Solomon Islands strains are undetectable by the PCR used in many studies and by health ministries. This assay is therefore not adequate for the eradication program. Next-generation genome sequence data are essential for these efforts.
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macrolide resistance in Treponema pallidum correlates with 23s rdna mutations in recently isolated clinical strains
Sexually Transmitted Diseases, 2016Co-Authors: Arbara J Molini, Sharo K Sahi, Laure C Tantalo, Charmie Godornes, Christina M Marra, Veronica Rodriguez, Stephanie L And, Mark C Fernandez, Sheila A LukehaAbstract:BackgroundHigh rates of 23S rDNA mutations implicated in macrolide resistance have been identified in Treponema pallidum samples from syphilis patients in many countries. Nonetheless, some clinicians have been reluctant to abandon azithromycin as a treatment for syphilis, citing the lack of a causal
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molecular typing of Treponema pallidum in ocular syphilis
Sexually Transmitted Diseases, 2016Co-Authors: Sara E Olive, Sharo K Sahi, Laure C Tantalo, Charmie Godornes, Robyn Neble Fanfai, Lauri E Markowitz, Sheila A Lukeha, Christina M MarraAbstract:BackgroundSyphilis can have many clinical manifestations, including eye involvement, or “ocular syphilis.” In 2015, an increase in reported cases of ocular syphilis and potential case clusters raised concern for an oculotropic strain of Treponema pallidum, the infectious agent of syphilis. Molecular
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reduced Treponema pallidum specific opsonic antibody activity in hiv infected patients with syphilis
The Journal of Infectious Diseases, 2016Co-Authors: Christina M Marra, Sharo K Sahi, Laure C Tantalo, Shelia Dunaway, Sheila A LukehaAbstract:BACKGROUND Human immunodeficiency virus (HIV)-infected individuals may have poorer serological responses to syphilis treatment and may be more likely to experience neurosyphilis. Treponema pallidum is cleared from sites of infection by opsonization, ingestion, and killing by macrophages. METHODS Serum samples from 235 individuals with syphilis were tested for T. pallidum-specific opsonic activity. Blood T. pallidum concentrations were determined by real-time polymerase chain reaction amplification of the tp0574 gene, and T. pallidum was detected in cerebrospinal fluid (CSF) by reverse-transcriptase polymerase chain reaction of 16S ribosomal RNA. RESULTS Opsonic activity was higher with higher serum rapid plasma reagin titers (P < .001), and in those treated for uncomplicated syphilis before serum collection (P < .001). Opsonic activity was lower in HIV-infected than in HIV-uninfected individuals even after the above factors were taken into account (P = .006). In participants in whom blood T. pallidum was detectable, those with the highest opsonic activity had lower blood T. pallidum concentrations. In multivariable analyses, there was not a significant relationship between opsonic activity and detection of T. pallidum in CSF or CSF-VDRL reactivity. CONCLUSIONS Serum T. pallidum-specific opsonic activity is significantly lower in HIV-infected individuals. Impaired T. pallidum-specific immune responses could contribute to differences in the course of disease or treatment response.
