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Journal of Clinical Microbiology, August 2009, p. 2510-2512, Vol. 47, No. 8
0095-1137/09/$08.00+0     doi:10.1128/JCM.00287-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Molecular Subtyping of Treponema pallidum subsp. pallidum in Lisbon, Portugal{triangledown}

R. Castro,1 E. Prieto,1 M. J. Águas,2 M. J. Manata,2 J. Botas,2 and F. Martins Pereira1*

Instituto de Higiene e Medicina Tropical, Unidade de Doenças Sexualmente Transmitidas, Universidade Nova de Lisboa,1 Hospital Garcia de Orta, Lisbon, Portugal2

Received 12 February 2008/ Returned for modification 20 April 2009/ Accepted 13 May 2009

The objectives of this study were to evaluate the reproducibility of a molecular method for the subtyping of Treponema pallidum subsp. pallidum and to discriminate strains of this microorganism from strains from patients with syphilis. We studied 212 specimens from a total of 82 patients with different stages of syphilis (14 primary, 7 secondary and 61 latent syphilis). The specimens were distributed as follows: genital ulcers (n = 9), skin and mucosal lesions (n = 7), blood (n = 82), plasma (n = 82), and ear lobe scrapings (n = 32). The samples were assayed by a PCR technique to amplify a segment of the polymerase gene I (polA). Positive samples were typed on the basis of the analysis of two variable genes, tpr and arp. Sixty-two of the 90 samples positive for polA yielded typeable Treponema pallidum DNA. All skin lesions in which T. pallidum was identified (six of six [100%]) were found to contain enough DNA for typing of the organism. It was also possible to type DNA from 7/9 (77.7%) genital ulcer samples, 13/22 (59.1%) blood samples, 20/32 (62.5%) plasma samples, and 16/21 (76.2%) ear lobe scrapings. The same subtype was identified in all samples from the same patient. Five molecular subtypes (subtypes 10a, 14a, 14c, 14f, and 14g) were identified, with the most frequently found subtype being subtype 14a and the least frequently found subtype being subtype 10a. In conclusion, the subtyping technique used in this study seems to have good reproducibility. To our knowledge, subtype 10a was identified for the first time. Further studies are needed to explain the presence of this subtype in Portugal, namely, its relationship to the Treponema pallidum strains circulating in the African countries where Portuguese is spoken.

* Corresponding author. Mailing address: Instituto de Higiene e Medicina Tropical, Rua da Junqueira, no. 96, Lisbon 1349-008, Portugal. Phone: 00351-213652600. Fax: 00351-213632105. E-mail: ritacastro{at}ihmt.unl.pt

{triangledown} Published ahead of print on 3 June 2009.

Journal of Clinical Microbiology, August 2009, p. 2510-2512, Vol. 47, No. 8
0095-1137/09/$08.00+0     doi:10.1128/JCM.00287-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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