Nadjet Benhaddou

Evaluation of a PCR Test for Detection of Treponema pallidum in Swabs and Blood

ABSTRACT Syphilis diagnosis is based on clinical observation, serological analysis, and dark-field microscopy (DFM) detection of Treponema pallidum subsp. pallidum, the etiological agent of syphilis, in skin ulcers. We performed a nested PCR (nPCR) assay specifically amplifying the tpp47 gene of T. pallidum from swab and blood specimens. We studied a cohort of 294 patients with suspected syphilis and 35 healthy volunteers. Eighty-seven of the 294 patients had primary syphilis, 103 had secondary syphilis, 40 had latent syphilis, and 64 were found not to have syphilis. The T. pallidum nPCR results for swab specimens were highly concordant with syphilis diagnosis, with a sensitivity of 82% and a specificity of 95%. Reasonable agreement was observed between the results obtained with the nPCR and DFM methods (kappa = 0.53). No agreement was found between the nPCR detection of T. pallidum in blood and the diagnosis of syphilis, with sensitivities of 29, 18, 14.7, and 24% and specificities of 96, 92, 93, and 97% for peripheral blood mononuclear cell (PBMC), plasma, serum, and whole-blood fractions, respectively. HIV status did not affect the frequency of T. pallidum detection in any of the specimens tested. Swab specimens from mucosal or skin lesions seemed to be more useful than blood for the efficient detection of the T. pallidum genome and, thus, for the diagnosis of syphilis.

Clinical and Serologic Baseline and Follow-Up Features of Syphilis According to HIV Status in the Post-HAART Era

There is a lack of large studies appraising the effect of the human immunodeficiency virus (HIV) on the course of syphilis since the advent of highly active antiretroviral therapy (HAART). We aimed to appraise the effect of HIV on clinical and serologic features of syphilis at baseline and during follow-up in the post-HAART era. We designed a retrospective cohort study of consecutive syphilis cases, diagnosed between 2000 and 2007, in an academic venereal disease center. Data were collected using standardized medical forms. Patients were treated according to the European guidelines. Serologic failure was defined as either a 4-fold rise in Venereal Disease Research Laboratory (VDRL) titers 30-400 days posttreatment or a lack of 4-fold drop in VDRL titers at 270-400 days posttreatment. Among 279 syphilis cases with informative baseline clinical and serologic data, HIV infection was significantly associated with men having sex with men, French origin, multiple partners, lesser usage of condom, history of sexually transmitted disease, early syphilis, anal primary chancre, and cutaneous eruption. Median baseline titer from the Treponema pallidum hemagglutination assay (TPHA) was higher in HIV-infected patients (p = 0.02). Among 144 informative syphilis cases, there was a nonsignificant trend for a lower rate of serologic response among HIV-positive patients (91.8% vs. 98.3%, p = 0.14). Serologic failure was significantly associated with a history of previous syphilis (p < 0.05). The median delay to serologic response was similar in HIV-positive (117 d) and in HIV-negative (123 d) patients (p = 0.44). We conclude that for patients under HAART treatment, the effect of HIV on serologic response to syphilis treatment is likely minimal or absent. Abbreviations: CSF = cerebrospinal fluid, FTA-abs = fluorescent treponemal antibody absorption test, HAART = highly active antiretroviral therapy, HIV = human immunodeficiency virus, HR = hazard ratio, MSM = men having sex with men, RPR = rapid plasma reagin, STD = sexually transmitted disease, TPHA = Treponema pallidum hemagglutination assay, VDRL = Venereal Disease Research Laboratory.

Molecular subtyping of Treponema pallidum in Paris, France.

Two major Treponema pallidum subtypes, 14 d/g and 14 d/f, were identified in a population of 119 patients with syphilis in Paris, France, characterized by a high proportion of men who have sex with men. A new subtype named 11 q/j was characterized, and a reinfection case was determined in 1 patient having consecuitve syphilis infection at 19-month interval.

The epidemiological and clinical presentation of syphilis in a venereal disease centre in Paris, France. A cohort study of 284 consecutive cases over the period 2000-2007

Since 2000, the incidence of syphilis has risen in developed countries. An updated knowledge of syphilis features is the key for early diagnosis and treatment. Our objective was to appraise the clinical and epidemiological presentation of syphilis in Paris, France. A retrospective monocentric descriptive study of 284 consecutive syphilis cases was conducted in a venereal disease centre (Paris, France), over the period 2000-2007. Epidemiological, clinical and microbiological data were systematically collected, using standardized medical forms. Overall, 95% of the cases occurred in men (271/284), 83% in men having sex with men (MSM) (231/278), 58% in patients having more than 10 partners/year (138/240) and 19% in patients who never use a condom (49/253). At least one STD has been previously diagnosed in 79% (220/279) of the cases. In 50.5% of the cases (142/281), HIV serology was positive. Most patients had primary (82/279, 29%) or secondary (125/279, 45%) syphilis. The most frequent physical signs in primary and secondary syphilis were, respectively, a genital chancre (63/82, 77%) and a diffuse exanthema (108/125, 86%). Syphilis occurs chiefly in MSM, often in HIV-positive patients. Many patients never use condoms. These data will help provide the basis for the development of national information and screening campaigns.

Human Treponema pallidum 11q/j isolate belongs to subsp. endemicum but contains two loci with a sequence in TP0548 and TP0488 similar to subsp. pertenue and subsp. pallidum, respectively

Background Treponema pallidum subsp. endemicum (TEN) is the causative agent of endemic syphilis (bejel). An unusual human TEN 11q/j isolate was obtained from a syphilis-like primary genital lesion from a patient that returned to France from Pakistan. Methodology/Principal findings The TEN 11q/j isolate was characterized using nested PCR followed by Sanger sequencing and/or direct Illumina sequencing. Altogether, 44 chromosomal regions were analyzed. Overall, the 11q/j isolate clustered with TEN strains Bosnia A and Iraq B as expected from previous TEN classification of the 11q/j isolate. However, the 11q/j sequence in a 505 bp-long region at the TP0488 locus was similar to Treponema pallidum subsp. pallidum (TPA) strains, but not to TEN Bosnia A and Iraq B sequences, suggesting a recombination event at this locus. Similarly, the 11q/j sequence in a 613 bp-long region at the TP0548 locus was similar to Treponema pallidum subsp. pertenue (TPE) strains, but not to TEN sequences. Conclusions/Significance A detailed analysis of two recombinant loci found in the 11q/j clinical isolate revealed that the recombination event occurred just once, in the TP0488, with the donor sequence originating from a TPA strain. Since TEN Bosnia A and Iraq B were found to contain TPA-like sequences at the TP0548 locus, the recombination at TP0548 took place in a treponeme that was an ancestor to both TEN Bosnia A and Iraq B. The sequence of 11q/j isolate in TP0548 represents an ancestral TEN sequence that is similar to yaws-causing treponemes. In addition to the importance of the 11q/j isolate for reconstruction of the TEN phylogeny, this case emphasizes the possible role of TEN strains in development of syphilis-like lesions.

Treponema pallidum 11qj Subtype May Correspond to a Treponema pallidum Subsp. Endemicum Strain.

Unusual 11qj subtype from 1 isolate was identified in a population of syphilis patients in Paris.1 Later on, Mikalova et al.2 suggested that this 11qj subtype may belong to a Treponema pallidum subsp. pertenue (TPE) rather than a T. pallidum subsp. pallidum (TPA) strain and was further commented by Lukehart and Giacani.3 Pathogenic treponemal strains (TPA, TPE, T. pallidum subsp. endemicum [TEN]) are morphologicaly similar, undistinguishable by serology and exhibit important gene identity.

Treponema pallidum induces systemic TH17 and TH1 cytokine responses.

ejd.2012.1841 Auteur(s) : Celine Bernardeschi1, Philippe A. Grange2, Michel Janier3, Ludivine Gressier1, Pierre L. Dion3, Nadjet Benhaddou4, Anne Bianchi5, Francois Lassau3, Marie-Francoise Avril1, Frederic Batteux6,a, Nicolas Dupin1,2,a nicolas.dupin@cch.aphp.fr 1 Service de dermatologie-venereologie, Hopital Cochin, 75006 Paris, France 2 Laboratoire de recherche en dermatologie, Universite Paris Descartes, 75679 Paris, France 3 Centre des MST, Hopital Saint-Louis, Paris 4 Service de [...]

Multi-locus sequence typing of Treponema pallidum subsp. pallidum present in clinical samples from France: Infecting treponemes are genetically diverse and belong to 18 allelic profiles

Treponema pallidum subsp. pallidum, the causative agent of sexually transmitted syphilis, detected in clinical samples from France, was subjected to molecular typing using the recently developed Multilocus Sequence Typing system. The samples (n = 133) used in this study were collected from 2010–2016 from patients with diagnosed primary or secondary syphilis attending outpatient centers or hospitals in several locations in France. Altogether, 18 different allelic profiles were found among the fully typed samples (n = 112). There were five allelic variants identified for TP0136, 12 for TP0548, and eight for TP0705. Out of the identified alleles, one, seven, and three novel alleles were identified in TP0136, TP0548, and TP0705, respectively. Partial allelic profiles were obtained from 6 samples. The majority of samples (n = 110) belonged to the SS14-like cluster of TPA isolates while 7 clustered with Nichols-like isolates. Patients infected with Nichols-like samples were more often older (p = 0.041) and more often diagnosed with secondary syphilis (p = 0.033) compared to patients infected with SS14-like samples. In addition, macrolide resistance caused by the A2058G mutation was found to be associated with allelic profile 1.3.1 or with strains belonging to the 1.3.1 lineage (p<0.001). The genetic diversity among TPA strains infecting the European population was surprisingly high, which suggests that additional studies are needed to reveal the full genetic diversity of TPA pathogens infecting humans.

Syphilis et grossesse

Syphilis is a sexually transmitted disease responsible for a congenital severe infection. Congenital syphilis is complicated by fetal loss/neonatal death (50%), prematurity (25%) and major long term sequelae in other surviving children (20%). Every woman delivering a stillborn after 20WG should be tested for syphilis. Early screening is the cornerstone of prevention, and should be repeated in women at higher risk of contamination. Maternal management relies on early benzathine penicillin administration. Neonatal management relies on early diagnosis and prompt adequate penicillin therapy.