Chlamydia trachomatis serovar distribution and other concurrent sexually transmitted infections in heterosexual men with urethritis in Italy.

The distribution of Chlamydia trachomatis serovars among 157 heterosexual male patients with urethritis and the presence of coinfections with other sexually transmitted infections were studied. One hundred seventeen (74.5%) patients, with a mean age of 33.7 years, were Italians, whereas 40 (25.5%) were immigrants coming from eastern European countries, Africa, and South America. All the immigrants and 82 (70.0%) Italian patients reported sex with prostitutes. Out of 157 patients, 73 (46.5%) were found positive for C. trachomatis in urethral secretions and eight different C. trachomatis serovars were identified. The most common serovars were E (n = 18; 24.7%), D (n = 15; 20.5%), G (n = 14;19.2%), and F (n = 12; 16.4%). The sequencing data showed a high degree of conservation of the omp1 gene. Thirty-six (46.7%) out of the 73 C. trachomatis-positive patients were coinfected with another sexually transmitted infection. The most common coinfection was gonorrhoea detected in 22 (30.1%) patients, followed by condyloma in eight (8.2%) patients, syphilis in five (6.8%), and HIV in three (4.1%).

Antenatal syphilis serology in pregnant women and follow-up of their infants in northern Italy.

Positive syphilis serology was noted in 119 (0.49%) of the 24 053 pregnant women delivering at St Orsola Hospital in Bologna, Italy, from November 2000 through July 2007. Six presumptive cases of congenital syphilis with IgM western blot positive results were found. Two infants had a positive cerebrospinal fluid (CSF) Venereal Disease Research Laboratory test result (one also had a positive CSF PCR result), another presented long-bone lesions, and the remaining three were preterm. These observations confirmed that antenatal syphilis screening facilitates treatment during pregnancy and offsets vertical transmission; moreover, the use of IgM western blot and careful CSF examination allowed the identification and treatment of high-risk newborns.

Borrelia burgdorferi VlsE antigen for the serological diagnosis of Lyme borreliosis.

In this study, raising and development of antibody response to Borrelia burgdorferi infection in 66 Italian patients suffering from culture-confirmed Lyme borreliosis erythema migrans (EM) was investigated. Sixty-two of 66 cultures obtained from biopsies were identified as B. afzelii by PCR. A total of 175 serially collected serum samples were tested by using two different sets of commercial assays: Enzygnost Lyme link VlsE/IgG and Enzygnost Borreliosis IgM (DADE Behring, Marburg, Germany) and LIAISON Borrelia IgG and IgM (Diasorin, Saluggia, Italy). Considering only samples obtained at first presentation when EM was clinically evident, 49/66 patients (72.4%) were IgG or IgM positive by Enzygnost, whereas 33/66 (50.0%) patients were IgG or IgM positive by LIAISON. Taking into account the follow-up period, eight patients sero-converted for IgG or IgM by Enzygnost and four by LIAISON. Similar and very good specificity values were obtained by all methods. Testing sera obtained from blood donors (n = 300) and from patients suffering from some of the most common biological conditions possibly resulting in false-positive reactivity in Lyme disease serology (n = 100) showed that Enzygnost Lyme link VlsE/IgG was the more specific (98.3%), followed by LIAISON Borrelia IgG (96.5%), and considering IgM tests, Enzygnost Borreliosis IgM showed to be 95.3%% specific, whereas the LIAISON Borrelia IgM was 92.8% specific. Recombinant VlsE antigens obtained from all three B.burgdorferi genospecies pathogenic to humans (included in Enzygnost Lyme link VlsE/IgG) greatly improved serodiagnosis of Lyme disease.

Development of a hamster model of Chlamydophila pneumoniae infection.

The aim of this study was to develop a new experimental model of Chlamydophila pneumoniae infection in the hamster. Intraperitoneal injection of C. pneumoniae purified elementary bodies (EBs) in the hamsters caused a systemic infection, since it was possible to isolate viable chlamydiae from several organs up to 14 days after infection. In particular, spleen infection was detectable up to 7 days post infection in 100% of animals. In contrast, cultures of the organs obtained from intranasally infected animals were far less frequently positive. Systemic infection probably occurred via macrophages, as demonstrated by the presence of intracellular chlamydial inclusions in peritoneal macrophages of peritoneally inoculated animals four days after infection. Furthermore, by infecting LLC-MK2 cells with supernatant preparations obtained from these macrophages, it was possible to observe the development of chlamydial intra-cytoplasmic inclusions after 96 h. Immunization of 18 hamsters with heat-inactivated purified EBs completely protected 16 animals and substantially reduced infection levels in the remaining two. Sera obtained from immunized hamsters prior to challenge reacted mainly against two C. pneumoniae proteins of about 60 kDa, when tested by immunoblot.

[Tick borne zoonosis: selected clinical and diagnostic aspects]. / Infezioni zoonosiche trasmesse da zecche: aspetti clinici e diagnostici in medicina umana.

Tick-borne zoonotic infections are among the most diffuse vector borne diseases: these large group of infections is caused by different microorganisms: Babesia spp., Borrelia spp., Rickettsia spp., Ehrlichia spp., Francisella tularensis, Coxiella burnetii) and tick-borne encephalitis virus. Babesiosis is caused by the protozoa (sporozoa) Babesia microti and it is quite rare in humans in Europe. The ixodids ticks are the competent vectors. A few symptomatic cases have been reported, mainly in splenectomized patients. The laboratory diagnosis is made by the microscopic identification of the parasites within the red blood cells in blood smears. The serologic diagnosis, based mainly upon IFA and WB techniques has only an epidemiological interest. Lyme borreliosis (Lyme disease) has been recognized as the most frequent vector borne disease in mild climate areas. The etiologic agent is a spirochete, belonging to the Borrelia burgdorferi sensu lato complex: B. burgdorferi sensu stricto, B. garinii and B. afzelii. Several additional species of this geno-complex have been identified but their pathogenic capability for humans still needs to be elucidated. Lyme borreliosis is clinically divided into three different clinical stages: the early disease, the disseminated infection and the persistent infection. Individual stages are caused by the diffusion of the spirochetes to different anatomic districts of the body. The main clinical symptoms are, for each stage: the erythema chronicum migrans in the early infection, the peripheral nerves and joint involvement in disseminated diseases and the acrodermatitis chronica atrophica (ACA) with central nervous system involvement in the late disseminated infection. The microbiological diagnosis is achieved by serologic techniques (IFA, EIA, WB) and by isolation of the spirochetes (in vitro culture and DNA amplification methods). Tick-borne relapsing fever (TBRF) is occasionally transmitted to humans by the soft ticks Ornithodorus and is caused by Borrelia spp. Different borreliae are responsible for TBRF in various geographic areas. The laboratory diagnosis is based upon the identification of spirochetes in peripheral blood by microscopic observation of Giemsa stained smears. Rickettsiosis diseases are caused worldwide by the obligate intracellular bacteria belonging to the genus Rickettsia. In the Mediterranean area the most frequently identified rickettsia is R. conorii, that causes the so called Mediterranean spotted fever. The serologic detection of a specific antibody response by IFA techniques is the most prominent tool for the diagnosis. In addition, the PCR method can be applied. Bacteria of the genus Ehrlichia are well known pathogens in veterinary medicine. Since the last decade their zoonotic capability has emerged and E. chafeensis, E. canis and the so called human granulocytic agent (HGE) have been identified in human diseases following a tick bite. The ehrlichiosis is characterized, in human, by a mild fever associated with lymphoadenopathy. The diagnosis is made on the identification of morulae (the intracytoplasmatic inclusion of the growing rickettsiae) in the white cells of peripheral blood. In addition the molecular diagnosis is also possible by PCR. Tick-borne encephalitis (TBE) is the only viral arthropod-borne encephalitis in Europe: it is caused by a flavivirus and it can also be transmitted by the ingestion of goat raw milk. The more relevant epidemiological figure is limited to the Alps, in particular to the Northern side (Austria). Isolated cases have been reported also in Italy. TBE is a benign self-limiting illness that usually recovers without any reliquate. The laboratory diagnosis is obtained by isolating the virus in cell cultures from the CSF or blood of acute phase patients. Serology is anyway the main laboratory tool to perform this diagnosis. Complement fixation and EIA IgM are the most used methods: the latter technique is particularly sensitive in early infection.

Comparative in vitro activity of garenoxacin against Chlamydia spp.

The in vitro susceptibilities of 33 isolates of Chlamydia trachomatis, Chlamydia pneumoniae and Chlamydia psittaci to a new quinolone drug, garenoxacin (BMS-284756), in comparison with levofloxacin, ciprofloxacin, doxycycline, erythromycin and roxithromycin, were determined. Garenoxacin was the most active of the quinolone drugs tested, with identical MIC and MBC, which ranged from 0.007 to 0.03 mg/L. The MIC and MBC of the other two quinolones tested, levofloxacin and ciprofloxacin, were also identical, ranging from 0.25 to 2 mg/L. The MICs and MBCs of doxycycline, erythromycin and roxithromycin were also determined.

Evaluation of recomWell Treponema, a novel recombinant antigen-based enzyme-linked immunosorbent assay for the diagnosis of syphilis.

OBJECTIVE: To evaluate the diagnostic performance of an enzyme immunosorbent assay (recomWell Treponema) for the diagnosis of syphilis. The novel recombinant antigens Tpn47, TpN17 and TpN15 were utilized. METHODS: A total of 782 human serum specimens, belonging to four different categories (blood donors, n = 200; routine laboratory screening for syphilis, n = 400; syphilis patients, n = 122; potential cross-reactors, n = 60), were evaluated to compare the sensitivity and specificity of the recomWell Treponema kit with a standard whole Treponema pallidum cell lysate antigen-based ELISA (Syphilis Screening) and with micro-haemagglutination (MHA-TP). RESULTS: The overall specificity and sensitivity of the recomWell Treponema IgG was 98.9% and 98.3%, respectively. The specificity and sensitivity of Syphilis Screening ELISA was 98.7% and 98.3%, respectively. The agreement between recomWell Treponema and Syphilis Screening was 100%, 97.8%, 95.9% and 95% among the blood donor specimens, screening samples, syphilis specimens and the potential cross-reactors, respectively. Values of concordance varying from 96.7% to 98.3% were found in the different groups of sera between recomWell Treponema and MHA-TP. In addition, recomWell Treponema demonstrated a good diagnostic performance when used to detect the IgM to T. pallidum. No false-positive sera were identified and, in 17/19 samples from primary infection, an IgM immune response was found. CONCLUSIONS: recomWell Treponema was shown to be a highly specific and sensitive method in all stages of syphilis screening and it can be considered as alternative to other ELISA tests based on native antigen preparations.

Western immunoblotting with five Treponema pallidum recombinant antigens for serologic diagnosis of syphilis.

Five immunodominant Treponema pallidum recombinant polypeptides (rTpN47, rTmpA, rTpN37, rTpN17, and rTpN15) were blotted onto strips, and 450 sera (200 from blood donors, 200 from syphilis patients, and 50 potentially cross-reactive) were tested to evaluate the diagnostic performance of recombinant Western blotting (recWB) in comparison with in-house whole-cell lysate antigen-based immunoblotting (wclWB) and T. pallidum hemagglutination (MHA-TP) for the laboratory diagnosis of syphilis. None of the serum specimens from blood donors or from potential cross-reactors gave a positive result when evaluated by recWB, wclWB, or MHA-TP. The evaluation of the immunoglobulin G immune response by recWB in sera from patients with different stages of syphilis showed that rTmpA was the most frequently identified antigen (95%), whereas only 41% of the specimens were reactive to rTpN37. The remaining recombinant polypeptides were recognized as follows: rTpN47, 92.5%; rTpN17, 89.5%; and rTpN15, 67.5%. The agreement between recWB and MHA-TP was 95.0% (100% with sera from patients with latent and late disease), and the concordance between wclWB and MHA-TP was 92.0%. The overall concordance between recWB and wclWB was 97.5% (100% with sera from patients with secondary and late syphilis and 94.6 and 98.6% with sera from patients with primary and latent syphilis, respectively). The overall sensitivity of recWB was 98.8% and the specificity was 97.1% with MHA-TP as the reference method. These values for sensitivity and specificity were slightly superior to those calculated for wclWB (sensitivity, 97.1%, and specificity, 96.1%). With wclWB as the standard test, the sensitivity and specificity of recWB were 98.9 and 99.3%, respectively. These findings suggest that the five recombinant polypeptides used in this study could be used as substitutes for the whole-cell lysate T. pallidum antigens and that this newly developed recWB test is a good, easy-to-use confirmatory method for the detection of syphilis antibodies in serum.

Eruptive pseudoangiomatosis.

Eruptive pseudoangiomatosis (EPA) is a rare, benign, spontaneously regressing childhood exanthem. The term was recently coined by Prose et al.1 to describe a dermatosis characterized by the sudden onset of a few to several bright red angioma-like papules with a different histopathology from the true angiomas. We describe three patients with the typical lesions of EPA but with some peculiar features not previously described. We discuss the suspected viral aetiology of EPA, and hypothesize a multifactorial aetiopathogenesis.

Uptake and killing of Leptospira interrogans and Borrelia burgdorferi, spirochetes pathogenic to humans, by reticuloendothelial cells in perfused rat liver.

In situ-perfused rat livers were infused with a single dose of 1.5 x 10(7) radiolabeled cells of Leptospira interrogans serovar icterohaemorrhagiae, the agent of leptospirosis, or with Borrelia burgdorferi IRS, the agent of Lyme disease. Significant (P<0.0001) differences in the liver uptake of L. interrogans and of B. burgdorferi were observed, the uptakes being 37.4%+/-2.3% for L. interrogans and 60.5%+/-3.1% for B. burgdorferi. Leptospires, in contrast to borreliae, were recovered from the livers when liver samples were cultured in growth medium. Leptospires but not borreliae were recovered in bile within 30 min of infusion. The association of leptospires and borreliae with reticuloendothelial cells of the liver was demonstrated by immunohistochemistry. Leptospires and borreliae were found to be associated with vimentin-positive cells and not with desmin-positive cells. Few leptospires but no borreliae were also seen associated with vimentin- and desmin-negative cells, suggesting the presence of leptospires outside the sinusoidal spaces, in the liver parenchyma.

Treponema pallidum surface immunofluorescence assay for serologic diagnosis of syphilis.

A surface immunofluorescence assay (SIFA) using live spirochetes was analyzed and compared with Western blot (WB), fluorescent treponemal antibody absorption (FTA-ABS), microhemagglutination (MHA-TP), and Treponema pallidum immobilization (TPI) assays for detecting serum antibodies to T. pallidum in patients with syphilis, in disease controls, and in healthy subjects. SIFA and WB were 99% sensitive (99 of 100 positive specimens) and specific (140 of 140 negative specimens); FTA-ABS showed a sensitivity and a specificity of 90 and 89% (90 of 100 positive and 125 of 140 negative specimens), respectively. MHA-TP showed a sensitivity of 84% (84 of 100 positive specimens) and a specificity of 98.5% (138 of 140 negative specimens). Finally, TPI had a sensitivity of 52% (52 of 100 positive specimens) and a specificity of 100% (140 of 140 negative specimens). The T. pallidum SIFA was therefore highly specific, showing no equivocal reactivities with control sera, and sensitive. The results suggest the possible use of SIFA as a confirmatory test in the serologic diagnosis of syphilis.