In this study, polymerase chain reaction (PCR) techniques were used to detect Treponema pallidum DNA in samples from patients with latent syphilis. Sixty-nine patients with latent syphilis and 18 with treated syphilis were included. Whole blood, plasma, sera and ear scrapings, totalling 235 samples from patients with latent syphilis, were obtained. Three PCR assays (47-PCR, polA-PCR and M-PCR assays) were performed. The 47-PCR yielded the highest number of positive samples -92/235 (39.1%), followed by M-PCR -90/235 (38.3%) and polA-PCR -73/235 (31.1%). Ear scrapings presented the highest number of positives (47/84 -56%), followed by plasma samples (36/84 -42.9%), whole blood (32/84 -38.1%) and sera (21/84 -25%). In conclusion, we have confirmed that T. pallidum can be found in blood of patients with latent syphilis. The 47-PCR technique was found to be the most sensitive, whereas ear lobe scrapings seem to be the best specimen for detection of T. pallidum DNA in latent syphilis.
In the present study, the performance of an enzyme-linked immunosorbent assay (ELISA) technique (Eti-syphilis-G and Eti-syphilis-M; DiaSorin) for detection of Treponema pallidum immunoglobulin M (IgM) and IgG antibodies for the laboratory diagnosis of syphilis was evaluated. Four hundred forty-one samples were studied. The sensitivity and specificity of the ELISA were 100 and 93%, respectively, compared with the results of a microhemagglutination assay for Treponema pallidum (MHA-TP) and 99.4 and 100%, respectively, compared with the results of the fluorescent treponemal antibody absorption (FTA-Abs) test. The results of the ELISA technique were concordant with those of MHA-TP for 98% of the samples tested, while the rate of concordance with the FTA-Abs test was 99.5%. The sensitivities of the rapid plasma reagin (RPR) test, MHA-TP, and the ELISA in the different phases of syphilis compared with the results of the FTA-Abs test were 92, 88, and 100%, respectively, for patients with primary syphilis; 100% for all tests evaluated for patients with secondary syphilis; 97.2, 99.4, and 100%, respectively, for patients with latent syphilis; and 57.9, 92.6, and 97.9%, respectively, for patients with past treated syphilis. The RPR test was reactive with 12 samples that were negative by all the specific tests. IgM antibodies were most frequently detected by the ELISA for IgM antibodies (32.8%) than by the FTA-Abs for IgM antibodies (28.4%). Detection of these antibodies by the FTA-Abs test and the ELISA for IgM antibodies decreased with the stage of disease (72 and 88%, respectively, for patients with primary syphilis to 17 and 19%, respectively, for patients with early latent syphilis). The high sensitivity and specificity of this ELISA technique during all stages of syphilis, together with the fact that it is a simple, objective, and easily automated method, lead us to believe that it could be used as a screening test for syphilis.
A molecular system was used to subtype Portuguese Treponema pallidum clinical strains isolated from both skin lesions and blood. The study with this system constitutes the first typing study in a European country. Three T. pallidum subtypes were found: subtypes 14a (50%), 14d (45.2%), and 14f (4.8%). Further studies are needed to better characterize the isolates involved in syphilis outbreaks.Syphilis, caused by the spirochete Treponema pallidum subsp. pallidum, is a multistage disease with a wide spectrum of clinical manifestations (8). Of particular relevance to public health is the recognition that syphilis increases the risk of transmission and acquisition of the human immunodeficiency virus (6). Until the end of the 20th century, the in vitro noncultivable condition of T. pallidum, allied with the high GC content of major portions of its genome (5), prevented the success of any strategy for the typing of T. pallidum clinical isolates. In 1998, Pillay et al. (13) made a very important step toward obtaining an understanding of the molecular epidemiology of T. pallidum by developing the only existing method for the genotyping of this pathogen. The method is based on the intrastrain variability of the acidic repeat protein gene (arp) and the Treponema pallidum repeat gene (tpr). To date, the laboriousness and low sensitivity of this procedure have limited its application to only five published studies (11,(13)(14)(15)17), focused mainly in two countries, South Africa and the United States. Consequently, there are only 288 typed T. pallidum strains worldwide (11,(13)(14)(15)17), which is a strikingly small number, considering the predicted syphilis incidence rate (18). In Portugal, early and congenital syphilis require mandatory notification (4); nonetheless, as in the rest of Europe, there is a complete lack of awareness of the diversity of the circulating T. pallidum strains. Here we report the results of a pioneer study in which we used the recent subtyping system developed by Pillay et al. at the Centers for Disease Control and Prevention (13) in order to identify and differentiate Portuguese T. pallidum isolates.Four hundred sixteen individuals (104 women and 312 men) suspected of having early syphilis (on the basis of clinical data and serology), most of whom were attending the major Portuguese sexually transmitted disease clinic (located at the Lapa Health Centre in Lisbon, Portugal), were evaluated for T. pallidum infection between 2004 and 2007. T. pallidum DNA was extracted from skin lesions or blood samples by using a QIAamp DNA mini kit (Qiagen, Valencia, CA), according to the manufacturer's instructions. T. pallidum was detected in 86 specimens (35 primary and 7 secondary lesions and 44 blood samples) by using a commercially available real-time PCR assay (Sacace, Caserta, Italy) that targets the gene coding for a recognized T. pallidum surface antigen, the 39-kDa basic membrane protein (bmp; locus TP1016 relative to the sequence of T. pallidum strain Nichols, GenBank accession number NC_000919 [3,5]...
SIGNIFICANCEHuman papillomavirus infection is highly prevalent in sexually active population and has been associated with anal and oropharyngeal cancers. This study estimates the prevalence of human papillomavirus on anal and oral samples from men and women with external anogenital warts. We found a high prevalence of human papillomavirus on extragenital sites (anal canal and oral mucosa) among patients with external anogenital warts. Both anal and oral human papillomavirus infections were more common in men who have sex with men than in heterosexual men. Anal highrisk HPV types (high risk as carcinogenic) were more common in women and in men who have sex with men. Overall, we demonstrated a high human papillomavirus burden in this population beyond anogenital warts, suggesting a greater impact of human papillomavirus vaccination.Human papillomavirus (HPV) infection is highly prevalent in the sexually active population. This study estimates the prevalence of HPV DNA in anal and oral samples from a cohort of men and women with incident anogenital warts. Anal and/or oral samples from 541 patients with anogenital warts were tested for 35 HPV genotypes using a PCR assay. The overall prevalence of anal HPV and oral HPV DNA was 59.9% (n = 305/509; 95% confidence interval (CI) 55.6-64.1%) and 14.5% (n = 78/538; 95% CI 11.8-17.7%), respectively. Among patients with perianal warts, the anal HPV DNA prevalence was 92.3% (95% CI 87.0-95.5%). Anal HPV DNA prevalence in patients with genital warts but no perianal warts was 55.7% (95% CI 50.6-60.7%). Both anal and oral HPV infections were more common in men who have sex with men than in heterosexual men (90.4% versus 38.5% and 20.8% versus 11.8%, respectively). Anal high risk-HPV infection was more common in women (58.8%) and in men who have sex with men (67.7%). We found that anogenital warts represent a clinical marker for both anal and oral HPV infections, including anal high risk-HPV infections, particularly among women and men who have sex with men. ActaDVActaDV Advances in dermatology and venereology Acta Dermato-Venereologica C. Lisboa et al. 558 www.medicaljournals.se/acta ActaDV ActaDV Advances in dermatology and venereology Acta Dermato-Venereologica 559 High HPV prevalence on anal and oral samples from patients with anogenital warts Acta Derm Venereol 2019 ActaDV ActaDV Advances in dermatology and venereology Acta Dermato-Venereologica C. Lisboa et al. 560 www.medicaljournals.se/acta ActaDV ActaDV Advances in dermatology and venereology Acta Dermato-Venereologica 561 High HPV prevalence on anal and oral samples from patients with anogenital warts Acta Derm Venereol 2019 ActaDV ActaDV Advances in dermatology and venereology Acta Dermato-Venereologica C. Lisboa et al. 562 www.medicaljournals.se/acta
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