A review of Treponema pallidum Molecular Detection Techniques, which is the most effective





Syphilis; PCR; Diagnosis; Molecular biology.


Syphilis is a polymorphic, infectious and systemic disease, caused by the bacteria T. pallidum, considered a disease of compulsory notification. Early diagnosis is an important factor in its control, this is performed by combining clinical findings and laboratory tests. Molecular techniques help in the diagnosis, however, there is no consensus on which technique would is the most appropriate, because studies present divergences such as which gene should be used as target, what sequence of primer, extraction method and type of sample. This review sought to investigate, based on scientific evidence, the most effective molecular techniques in the detection of T. pallidum DNA. Therefore, a systematic review of the literature was conducted in the PubMed, Science Direct and Lilacs databases, using the descriptors "syphilis" and "PCR" and "diagnosis" and "molecular biology", selecting works published in the last ten years. The sample consisted of 10 articles. As for the study design, the clinical trial was 100% prevalent. As for the method used, variations were evidenced, 6 used Nested PCR, 5 real-time PCR and 4 conventional PCR, 5 used more than one variation. The difference between them, such as time to evaluate a reaction, chance of contamination, reproducibility, specificity, sensitivity, precision and diagnostic accuracy, showed that the two genes of greatest interest are pol A and tpp47. Molecular tests, in particular nested PCR, proved to be promising, with good sensitivity and specificity in the diagnosis of syphilis together with serology to avoid clinical complications and disease transmission.


Ageron, A G., Laurent, F., Schrenzel. J., Charton, B., Getaz, G. J., Tangomo, M., Tristan, B., Sednaoui, P., Lautenschlager, S., Trellu, LT., Tejada, B. M., Cavassini, M., Emonet, S., Perneger, T., Salord H. (2015). Perfonmance of the 47 kilodalton menbrane protein versus DNA Polymerase I genes for detection of Treponema pallidum by PCRin ulcers. Journal of Clinical Microbiology. Mar;53(3):976-80. DOI: 10.1128/JCM.03444-14

Ageron, A. G., Ninet, B., Trellu, L. T., Lautenschlager, S., Furrer, H., Piguet, V., Schrenzel, J., Hirschel, B. (2009). Assessment of a Real Time PCR to Diagnose Syphilis fron deversi biologicalsample. Sexually Transmitted Infections. Ago;85(4):264-9. DOI: 10.1136/sti.2008.034314

Brasil, M.d.S. (2016). Manual Técnico para o Diagnóstico da Sífilis. - PNCQ.

Brasil, M.d.S. (2019).Boletim Epidemiológico da Sífilis.

Brasil, M. D. S. (2015). Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecção Sexual Tranmissivel.

Brischetto, A., Gassiep, I., Whiley, D., Norton, R. (2018). Retrospective review of Treponema pallidum PCR and serology results: are both tests necessary? Journal of Clinical Microbiology. p.1-7, v.56 n.5. 56:e01782-17. DOI: 10.1128/JCM.01782-17.

Carlson, JA., Dabiri, G., Cribier, Sell, S. (2011). The immunopathobiology of syphilis: the manifestations and course of syphilis are determined by the level of delayed-type hypersensitivity. The American Journal of Dermatopathology. Jul;33(5):433-460. DOI: 10.1097/dad.0b013e3181e8b587

Castro MRTd. (2004). Contribuição para o estudo da infeçãopor Treponema pallidum subespecie pallidum: Resposta sorologica, Diagnsotico Molecular e Genotipagem. Dissertação de Doutorado. Lisboa - Portugal (LISBOA): Universidade Nova Lisboa.

Castro, R., Águas, M. J., Batista, T., Araújo, C., Mansinho, K., Pereira, FdaL. (2016). Detection of Treponema pallidum Sp. Pallidum DNA in Cerebrospinal Fluid (CSF) by Two PCR Techniques. Journal of Clinical Laboratory Analysis. Sep;30(5):628-32. DOI: 10.1002/jcla.21913. Epub 2016 Feb 18. PMID: 26892231; PMCID: PMC6807054.

Castro, R., Prieto, E., Aguas, M. J., Manata, M. J., Botas, J., Pereira F. M. (2009). Molecular subtyping of Treponema pallidum subsp. pallidum in Lisbon, Portugal. Journal of Clinical Microbiology. Aug;47(8):2510-2. DOI: 10.1128/JCM.00287-08. Epub 2009 Jun 3. PMID: 19494073; PMCID: PMC2725690.

Cohen, S. E., Klausner, J. D., Engelman, J., Philip, S. (2013). Syphilis in the modern era: an update for physicians. Infectious Disease Clinics of North America. Dec;27(4):705-22. DOI: 10.1016/j.idc.2013.08.005. PMID: 24275265.

Fraga, D. D. ., & Goldini, L. Z. (2013). Detecção do Treponema pallidum em Liquido Cefalorraquidiano LCR pela reação em cadeia de polimerase PCR em pacientes HIV positivos assintomaticos com diagnsotico de sífilis latente. Dissertação para titulo de Mestre. Porto Alegre: Universidade Federal do Rio Grande do Sul.

Galvão, C. M., Sawada, N. O., Trevizan, M. A. (2004). Revisão Sistemática: Recurso que proporciona a incorporação das evidências na prática da enfermagem. Revista Latino-americano Enfermagem, 12(3):549-56. DOI:10.1590/S0104-11692004000300014

Galvão, T. F., Pereira, M. G. (2014). Revisões sistemáticas da literatura: passos para sua elaboração. Epidemiologia e. Serviços de Saúde. 23 (1):183-184. DOI:10.5123/S1679-49742014000100018.

Glatz, M., Juricevic, N. Altwegg, M., Bruisten, S., Komericki, P., Lautenschlager, S., Weber, R., Bosshard, P. P. (2014). A multicenter prospective trial to asses a new real-time polymerase chain reaction for detection of Treponema pallidum, herpes simplex-1/2 and Haemophilus ducreyi in genital, anal and oropharyngeal ulcers. Clinical Microbiology Infection. Dec;20(12):O1020-7. DOI: 10.1111/1469-0691.12710. Epub 2014 Jul 25. PMID: 24909546.

Grange, P. A., Gressier, L., Dion, P. L., Farhi, D., Benhaddou, N., Gerhardt, P., Morini, J. P., Deleuze, J., Pantoja, C., Bianchi, A., Lassau, F., Avril, M. F., Janier, M., Dupin, N. (2012). Evaluation of a PCR test for detection of treponema pallidum in swabs and blood. Journal of Clinical Microbiology. Mar;50(3):546-52. DOI: 10.1128/JCM.00702-11. Epub 2012 Jan 4. PMID: 22219306; PMCID: PMC3295187.

Grange, P. A., Jary, A., Isnard, C., Burrel, S., Boutolleau, D., Touati, A., Bébéar, C., Saule, J., Martinet, P., Robert, J. L., Moulene, D., Vermersch-Langlin, A., Benhaddou, N., Janier, M., Dupin, N. (2021). Use of a Multiplex PCR Assay To Assess the Presence of Treponema pallidum in Mucocutaneous Ulcerations in Patients with Suspected Syphilis. Journal of Clinical Microbiology. Jan 21;59(2):e01994-20. doi: 10.1128/JCM.01994-20. PMID: 33177120; PMCID: PMC8111117.

Korenromp, E. L., Rowley, J., Alonso, M., Mello, MB., Wijesooriya, N. S.,7 Mahiné, SG., Ishikawa, N., Le, L. V., Newman-Owiredu, N., Nagelkerke, N., Newman, L., Kamb, M.,7 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 journal.pone.0211720. PLoS One. 14(7):1-17. DOI:10.1371/journal.pone.0219613.

Lafond, R. E., Lukehart, S. A. (2006). Biological basis for syphilis. Clinical Microbiology Reviews. Jan;19(1):29-49. DOI: 10.1128/CMR.19.1.29-49.2006. PMID: 16418521; PMCID: PMC1360276.

Liu, H., Rodes, B. Chen, C. Y., Steiner, B. (2001). New tests for syphilis: rational design of a PCR method for detection of Treponema pallidum in clinical specimens using unique regions of the DNA polymerase I gene. Journal of Clinical Microbiology. May;39(5):1941-6. DOI: 10.1128/JCM.39.5.1941-1946.2001. PMID: 11326018; PMCID: PMC88053.

Luo, Y., Xie, Y., Xiao, Y. (2021).Laboratory diagnostic tools for syphilis: Current status and future prospcts. Frontiers in cellular and infection microbiology. 8 de fevereiro de 2021;10:574806. DOI: 10.3389/fcimb.2020.574806. 33628742; PMCID: PMC7897658.

Noda, A. A., Rodríguez, I., Grillová, L., Bosshard, P. P., Lienhard, R. (2019). Accuracy of PCR and serological testing for the diagnosis of primary syphilis: Both tests are necessary. International Journal of STD & AIDS. 30: 1087–1094. DOI:10.1177/0956462419859764

Pereira, J. B.,7 Barbosa Júnior, W. L., Silva, E. D., Aquino, A. E. C. A., Oliveira, P. M. S., Melo, FL. (2019). Comparação de técnicas de extração de DNA de Treponema Pallidum para o diagnóstico molecular da sífilis. Brazilian Journal of Health Review. Curitiba, v. 2, n. 4, p. 3681-3697. DOI:10.34119/bjhrv2n4-131

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. 13 (23): 17-25.

Shukalek, C. B., Lee, B., Fathima, S., Chu, A., Fonseca, K.,7 Somayaji, R. (2021). Comparative Analysis of Molecular and Serologic Testing for Primary Syphilis: A Population-Based Cohort Study. Frontiers in Cellular and. Infection Microbiology. 11:579660. DOI: 10.3389/fcimb.2021.579660

Taylor, M. M., Li, W. Y., Skinner, J., Mickey, T. (2014). Viral loads among young HIV-infected men with early syphilis. Journal of the International Association Providers AIDS Care. Nov-Dec;13(6):501-5. DOI: 10.1177/2325957414536229. Epub 2014 Jun 4. PMID: 24899260; PMCID: PMC6754093.

Theel, E. S., Katz, S. S., Pillay, A. (2020). Molecular and direct detection Testes for Treponema pallidum subspecies pallidum: A review of the literature, 1964-2017. Clinical Infectious Diseases. Jun 24;71(Suppl 1):S4-S12. DOI: 10.1093/cid/ciaa176. 32578865; PMCID: PMC7312206.

Tipple, C., Hanna, M. O. F., Hill, S., Daniel, J., Goldmeie, D., McClure, M., Taylor, G. P. (2011). Getting the measure of syphilis: qPCR to better understand early infection. Sexually Transmited Infections p. 479-485, v. 87, n.6. DOI:10.1136/sti.2011.049494

Vrbová, E., Mikalová, L., Grillová, L., Pospíšilová, P., Strnadel, R., Dastychová, E., Kojanová, M., Kreidlová, M., Vaěousová, D., Rob, F.,& Procházka, P., Krchěáková, A., Vaškə, V., Woznicová, V., Dvořáková Herová, M., Kuklová, I., Zákoucká, H., Šmajs, D. (2020). A retrospective study on nested PCR detection of syphilis treponemes in clinical samples: PCR detection contributes to the diagnosis of syphilis in patients with seronegative and serodiscrepant results. PLos One. Ago 20;15(8):e0237949. DOI: 10.1371/journal.pone.0237949. 32817658; PMCID: PMC7446855.

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. May 9;7(1):83. DOI: 10.1038/s41426-018-0085-2. Erratum in: Emerging Microbes & Infections. 2020 Dec;9(1):1174. PMID: 29739928; PMCID: PMC5940865.

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

Zhang, Y., Dai, X., Ren, Z., Lin, H., Cao, W., Ye, X. (2019). A Novel Nested Real-time Polymerase Chain Reaction for Treponema pallidum DNA in Syphilis Biospecimens. Sexually Transmited Diseases. Jan;46(1):41-46. DOI: 10.1097/OLQ.0000000000000908. PMID: 30247262.



How to Cite

GRASSI, V. M. T.; GRASSI, L. T. .; ROSSETTI, M. L. R. A review of Treponema pallidum Molecular Detection Techniques, which is the most effective. Research, Society and Development, [S. l.], v. 10, n. 9, p. e59310918478, 2021. DOI: 10.33448/rsd-v10i9.18478. Disponível em: https://rsdjournal.org/index.php/rsd/article/view/18478. Acesso em: 25 sep. 2021.



Review Article