Detection of Treponema pallidum in blood samples fixed on card for diagnosis of congenital syphilis




Congenital syphilis; PCR; Diagnosis; Treponema pallidum.


Congenital syphilis (CS) is a vertically transmitted disease caused by Treponema pallidum causing an infection of the baby in the uterus. Diagnosis of CS is difficult and has been made by a combination of clinical and laboratory criteria, which makes it necessary to search for new diagnostic methods to aid decision making. Molecular tests, despite not being routine in laboratories, are being used more and more. The pre-analytical stage of sample collection, DNA extraction and preservation is essential. The use of fixed blood on a card (SFC) has been an alternative in the diagnosis of various diseases due to the ease of carrying out the pre-analytical stage. Our study aimed to detect T. pallidum by polymerase chain reaction (PCR) from CFS of newborns that had a positive PCR result with the cerebrospinal fluid (CSF) sample. . Eighty-two paired samples (blood and CSF) were analyzed from 41 NB at the Hospital da Criança Santo Antônio (HCSA), in Porto Alegre, RS, Brazil, 22 positive and 19 negative. Sensitivity and specificity were 63.3% and 100%, respectively. The agreement between the methods was substantial (K=0.619), showing that PCR directly from CFS can be promising, as long as it is improved, it is a possibility to diagnose CS and facilitate neonatal screening.


Ali, N., Bello, G. L., Rossetti, M. L. R., Krieger, M. A., & Costa, A. D. T. (2020). Demonstração de um protocolo rápido e fácil de amostra para resposta para rastreamento de tuberculose em ambientes de ponto de cuidado: Uma prova de estudo conceitual. PLoS ONE 15(12): e0242408.

Bankamp, B., Sein, C., Pukuta- Simbu, E., Anderson, R., Abernathy, E., Chen, M-H., Muyembe-Tamfum, J-J., Wannemuehler, K. A., Waku-Kouomou, D., Lopareva, E. N., Icenogle, J. P., Rota, P. A., & Goodson, J. L. (2019). Use of FTA cards to transport throat swabs and oral fluid samples for molecular detection and genotyping of measles and rubella viruses. Journal of Clinical Microbiology 57:e00048-19. .00048-19

Brasil, Boletim Epidemiológico da Sífilis. Ministério da Saúde, 2019

Brasil, Governo do. (2021). Teste do Pezinho será ampliado e detectará até 50 novas doenças.

Brasil. Ministério da Saúde, Secretaria de Vigilância em Saúde, Departamento de Vigilância, Prevenção e Controle das Doenças Sexualmente Transmissíveis, Aids e Hepatites Virais. Manual Técnico para Diagnóstico da Sífilis: Ministério da Saúde, 2016.

Castro, R., Aguas, M. J., Batista, T., Araujo, C., Mansinho, K., & Pereira, F. L. M. (2016). Detection of Treponema pallidum Sp. Pallidum DNA in Cerebrospinal Fluid (CSF) by to PCR Techniques. Journal of Clinical Laboratory Analysis. 30: 628-632. DOI 10.1002/jcla.21913

Focaccia R. Tratado de infectologia. (5a ed.), editora Atheneu; 2015.

Fraga, D., Muller, A. L., Czykiel, M. S., De Armas, Y., Wissmann, G., & Goldani, L. Z. (2014). Detection of Treponema pallidum by semi-nested PCR in the cerebrospinal fluid of asymptomatic HIV-infected patients with latent syphilis. Clinical Laboratory. 60(12):2051-2054. 10.7754/clin.lab.2014.140605

Fraser, C. M., Norris, S. J., Weinstock, G. M., White, O., Sutton, G. G., Dodson, R., & Sodergreen, E. (1988). Complete genome sequence of Treponema pallidum, the syphilis spirochete. Science. 281, 375-388. 10.1126/science.281.5375.375

Ghanem, K. G., Ram, S., & Rice, P. A. (2020). The Modern Epidemic of Syphilis. The New England Journal of Medicine. 382(9):845-854. 10.1056/NEJMra1901593.

Iared, W., Puchnick, A., Bancovsky, E., Bettini, P. R., Vedolin, L. M., & Chammas, M. C. (2018). Reproducibility of a quantitative system for assessing the quality of diagnostic ultrasound. Radiologia Brasileira. Mai/Jun;51(3):172–177. 10.1590/0100-3984.2017.0021.

Iroh Tam, P-Y., Hernandez-Alvarado, N., Schleiss, M. R., Hassan-Hanga, F., Onuchukwu, C., Umoru, D., & Obaro, S. K. (2016) Molecular Detection of Streptococcus pneumoniae on Dried Blood Spots from Febrile Nigerian Children Compared to Culture. PLoS ONE 11(3): e0152253.

Janier, M., Hegyi, V., Dupin, N., Unemo, M., Triplica, G. S., Potocnik, M., Frances, P., & Patel, R. (2014). European guideline on the management of syphilis. Journal of the European Academy of Dermatology and Venereology. 28(12):1581-1593. 10.1111/jdv.12734

Korenromp, E. L., Rowley, J., Alonso, M., Mello, M. B., Wijesooriya, N. S., Mahiané, S. G., Ishikawa, N., Le, L. V., Newman-Owiredo, M., Magalkerke, N., Newman, L., Broutet, N., & Taylor, M. M. (2019) Global burden of maternal and congenital syphilis and associated adverse birth outcomes, Estimates for 2016 and progress since 2012. PLoS ONE 14(2): e0211720.

Macedo, E. M. (2005) Há 100 anos, a descoberta do Treponema pallidum. Anais Brasileiros de Dermatologia. 80, 547-548.

Michelow, I. C., Wendel, G. D., Norgard, M. V., Zeray, F., Leos, N. K., Alsaadi, R., & Sánches, P. J. (2002). Central Nervous System Infection in Congenital Syphilis. The New England Journal of Medicine. 346(23):1792–8. 10.1056/NEJMoa012684.

Mota, C. A., Venazzi, E. A. S., Zanzarini, P. D., Aristides, S. M. A., Lonardoni, M. V. C., & Silveira, T. G. V. (2021). Filter paper performance in PCR for cutaneous leishmaniasis diagnosis. Revista da Sociedade Brasileira de Medicina Tropical. 54.

Palmer, H. M., Higgins, S. P., Herring, A. J., & Kingston, M. A. (2003). Use of PCR in the diagnosisof early syphilis in the inited Kingdom. Sexually Transmitted Infections. 79:479-83.

Pinilla, G., Campos, L., Durán, A., Navarrete, J., & Muñoz, L. (2018). Detección de Treponema pallidum subespecie pallidum para el diagnóstico de sífilis congénita mediante reacción en cadena de la polimerasa anidada. Biomedica. 38(1):128-35. 10.7705/biomedica.v38i0.3740. PMID: 29676865.

Pinilla, G. B., Chavarro, B. P., Moreno, M. A., Navarrete, J. O., & Muñoz, L. M. (2015). Determinación de los genes, 16S ADNr, polA, y TpN47, em la detección de Treponema pallidum subsp. pallidum para el diagnóstico de sífilis congénita. NOVA. 2015; 13 (23): 17-25.

Secretaria do estado de São Paulo. Guia de bolso- Manejo da sífilis em gestante e sífilis congênita. 2016. (2a ed.).

Theel, E. S., Katz, S. S., & Pillay, A. (2020). Molecular and Direct Detection Tests for Treponema pallidum Subspecies pallidum: A Review of the Literature, 1964–2017. Clinical Infectious Diseases. 71(Supplement_1):S4-S12. 10.1093/cid/ciaa176

Wang, C., Cheng, Y., Liu, B., Wang, Y., Gong, W., Qian, Y., Guan, Z., Lu, H., Gu, X., Shi, M., & Zhou, P. (2018). Sensitive detection of Treponema pallidum DNA from the whole blood of patients with syphilis by the nested PCR assay. Emerging Microbes Infections, 7:1, 1-7, 10.1038/s41426-018-0085-2 t.

Weinstock, G. M., Hardham, J. M., McLeod, M. P., Sodergren, E. J., & Norris, S. J. (1988). The genome of Treponema pallidum: New light on the agent of syphilis. FEMS Microbiol Rev. 1998;22(4):323-332.

Woods C. R. Syphilis in children: congenital and acquired. Seminars in Pediatric Infectious Diseases. 16(4):245-257.

Workowski, K. A., & Bolan, G. A. (2015). Centers for Disease Control and Prevention. Sexually transmitted diseases treatment guidelines, 2015. Morbidity and Mortality Weekly Report. Recommendations and Reports. 64(RR-03):1-137. Erratum in: MMWR Recomm Rep. 64(33):924.

World Health Organization Global adult estimates of chlamydia, gonorrhoea, trichomoniasis and syphilis including maternal and congenital syphilis, 2016, WHO; 2020, 76p.



How to Cite

GRASSI, V. M. T.; MELO, R. C. C.; GRASSI, L. T. .; SILVA, M. S. N. .; ROSSETTI, M. L. R. Detection of Treponema pallidum in blood samples fixed on card for diagnosis of congenital syphilis. Research, Society and Development, [S. l.], v. 10, n. 10, p. e511101019151, 2021. DOI: 10.33448/rsd-v10i10.19151. Disponível em: Acesso em: 30 sep. 2023.



Health Sciences