Wentao Chen1, David Šmajs2, Yongfei Hu1, Wujian Ke1, Petra Pospíšilová2, Kelly L Hawley3, Melissa J Caimano4, Justin D Radolf4,5, Arlene Sena6, Joseph D Tucker6,7, Bin Yang1, Jonathan J Juliano6,8,9, Heping Zheng1, Jonathan B Parr6,9. 1. Dermatology Hospital, Southern Medical University, Guangzhou, China. 2. Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic. 3. Division of Infectious Diseases and Department of Pediatrics, Connecticut Children's Medical Center, Hartford, Connecticut, USA. 4. Departments of Medicine, Pediatrics, and Molecular Biology and Biophysics, UConn Health, Farmington, Connecticut, USA. 5. Departments of Genetics and Genome Sciences, and Immunology, UConn Health, Farmington, Connecticut, USA. 6. Division of Infectious Diseases, Department of Medicine, University of North Carolina, Chapel Hill, North Carolina, USA. 7. Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom. 8. Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina, Chapel Hill, North Carolina, USA. 9. Curriculum in Genetics and Molecular Biology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina, USA.
Abstract
BACKGROUND: Whole-genome sequencing (WGS) of Treponema pallidum subspecies pallidum (TPA) has been constrained by the lack of in vitro cultivation methods for isolating spirochetes from patient samples. METHODS: We built upon recently developed enrichment methods to sequence TPA directly from primary syphilis chancre swabs collected in Guangzhou, China. RESULTS: By combining parallel, pooled whole-genome amplification with hybrid selection, we generated high-quality genomes from 4 of 8 chancre-swab samples and 2 of 2 rabbit-passaged isolates, all subjected to challenging storage conditions. CONCLUSIONS: This approach enabled the first WGS of Chinese samples without rabbit passage and provided insights into TPA genetic diversity in China.
BACKGROUND: Whole-genome sequencing (WGS) of Treponema pallidum subspecies pallidum (TPA) has been constrained by the lack of in vitro cultivation methods for isolating spirochetes from patient samples. METHODS: We built upon recently developed enrichment methods to sequence TPA directly from primary syphilis chancre swabs collected in Guangzhou, China. RESULTS: By combining parallel, pooled whole-genome amplification with hybrid selection, we generated high-quality genomes from 4 of 8 chancre-swab samples and 2 of 2 rabbit-passaged isolates, all subjected to challenging storage conditions. CONCLUSIONS: This approach enabled the first WGS of Chinese samples without rabbit passage and provided insights into TPA genetic diversity in China.
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