Raffaele Del Prete

Prevalence and genotypes identification of human papillomavirus infection in a population of South Italy

BACKGROUND A limited number of human papillomavirus (HPV) types account for the majority of invasive cervical cancer cases. OBJECTIVES To assess, in a southern Italian region, where HPV infection had not yet been investigated, the prevalence of type-specific HPV infection. STUDY DESIGN Multiplex PCR was used to test cervical specimens from 871 asymptomatic women. RESULTS The HPV infection rate was 23.1%, with the highest prevalence being observed in women aged 20-30 years (32.6%). Type 16 was the most frequent HPV type detected either in mono-infected (39.8%) or in multi-infected (46.3%) women. CONCLUSIONS The HPV infection rate was higher than reported from other Italian areas. Our results further emphasise the importance of vaccinations to immunize females before they acquire HPV infection.

Polymerase chain reaction detection of Bartonella henselae bacteraemia in an immunocompetent child with cat-scratch disease

Abstract A case of Bartonella henselae bacteraemia is reported in an immunocompetent 8-year-old boy with cat-scratch disease. Serology to B. henselae, diagnosed by polymerase chain reaction, was positive. DNA was extracted from peripheral whole blood and amplified with specific primers targeting the htrA gene of B. henselae. A non-isotopic hybridization assay with a species-specific oligonucleotide probe was used to detect the amplified product. Conclusion The polymerase chain reaction can be used for the rapid laboratory diagnosis of bacteraemia in cat-scratch disease.

Prevalence of antibodies to Bartonella henselae in patients with suspected cat scratch disease (CSD) in Italy

Cat scratch disease (CSD) is a relatively new diagnosed illness with clinical signs of self-limiting regional lymphadenopathy accompanied by symptoms of fever and malaise, to encephalopathy and neuropathy, occurring after a cat scratch or flea bite. Bartonella henselae is now accepted as the etiologic agent of CSD. From January 1994 to September 1998, 412 patients were evaluated for suspect CSD in Italy. Sera were tested for antibodies to B. henselae by a commercially available indirect immunofluorescent assay (IFA), based on B. henselae-infected Vero-cells as the antigen substrate. Of the 412 patients, 26 (6.3%) were considered positive having titers of immunoglobulin G (IgG) to B. henselae of 64 or higher. In these patients CSD was indeed confirmed by either histopathologic examination of lymph nodes biopsy or fourfold raise in antibody titers. Nevertheless, sera were tested by IFA for Afipia felis and one showed a double reactivity to B. henselae and A. felis. Finally, three sera, negative to B. henselae serology, were positive to A. felis. Three hundred and eighty-six patients received alternative diagnoses. One hundred and twenty-five serum samples from control subjects were negative by IFA for either B. henselae or A. felis. Moreover, a cross-reactivity with sera from patients affected by other diseases was not observed. Our study shows that the ascertained cases of CSD are etiologically determined by B. henselae, IFA assay is confirmed as a useful tool in the laboratory diagnosis and, over a 5 years period of study, the incidence of CSD in Italy has been low.

Preliminary evidence of endotoxic activity of Bilophila wadsworthia

In this study we describe two properties of the Gram-negative bacterium Bilophila wadsworthia, namely the ability to clot Limulus lysate and the capacity to induce the production of tissue factor-like procoagulant activity by human mononuclear cells in vitro. Although exhibited at a lower degree when compared with those of typical Gram-negative bacteria or Gram-negative endotoxin those activities may account in part for Bilophilas pathogenicity. The capacity indeed to induce fibrin formation through the interaction with mononuclear cells suggests one mechanism by which the microorganism might cause abscess formation in the host. Moreover, since this activity is dependent on the number of Bilophila interacting with mononuclear cells, we hypothesize that this biological activity is closely influenced by growth environment.

Rapid and sensitive detection of blaKPC gene in clinical isolates of Klebsiella pneumoniae by a molecular real-time assay

BackgroundThe aim of this study was the rapid identification of blaKPC gene in 38 Klebsiella pneumoniae clinical isolates with reduced susceptibility to carbapenems. The modified Hodge Test (MHT) was carried out to phenotypically determine whether resistance to carbapenems was mediated by a carbapenemase. The detection of the blaKPC gene was performed by real-time acid nucleic sequence-based amplification (NASBA™™), specifically designed for the detection of KPC RNA target.ResultsThirty-two/38 isolates evaluated by MHT showed the production of carbapenemases, while all the strains exhibited the production of KPC by inhibition test with phenylboronic acid (the combined disk test with IPM/IPM plus phenylboronic acid). The detection of blaKPC gene by Nuclisens EasyQ KPC yielded positive results in 38/38 (100%) strains. The presence of blaKPC gene was confirmed in all K. pneumoniae isolates when tested by the gold standard PCR assay.ConclusionsIn consideration of the serious challenge represented by infections due to K. pneumoniae it appears necessary the rapid identification of carbapenemases in clinical settings as it is made possible by the use of NASBA™ assay.

Tuberculous Otitis Media with Facial Paralysis: A Clinical and Microbiological Diagnosis—A Case Report

The tuberculosis of the ear is rare, and in most cases the clinical picture resembles that of a chronic otitis media. The diagnosis is often delayed, and this can lead to irreversible complications such as hearing loss and/or facial paralysis. In view of its rare occurrence, we report a case of primary tuberculous otitis media in a 87-year-old female patient. The diagnosis was made on the basis of both histological and microbiological findings. In particular, gene amplification techniques such as real-time polymerase chain reaction are useful method for rapid diagnosis and detecting tuberculous bacilli usually present at very low number. Early diagnosis is essential for the prompt institution of antituberculous therapy.

Simultaneous detection and identification of STI pathogens by multiplex Real-Time PCR in genital tract specimens in a selected area of Apulia, a region of Southern Italy

PurposeGenital tract infections are globally a major cause of morbidity in sexually active individuals. The aim of this study was to investigate the prevalence and associations of co-infections of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Mycoplasma hominis (MH), Mycoplasma genitalium, Ureaplasma urealyticum (UU) and Ureaplasma parvum (UP) in specimens collected from female (SF) and male (SM) patients.Methods1575 samples from 1575 individuals from the geographical area around Bari, Apulia region in Southern Italy, were collected and analyzed by a multiplex Real-Time PCR (mRT-PCR) (AnyplexTM II STI-7, Seegene, Inc., Seoul, Korea) assay.Results455/1575 (28.89%) samples resulted positive for at least one of the targets named above. Statistically significant differences in prevalence of the pathogens between SF and SM were not detected except for UP (24.92% in SF vs 8.91% in SM). Prevalence of co-infections was 6.84 and 3.96% in SF and SM, respectively. Moreover, MH presence in SF, but not in SM, was associated with UU and UP.ConclusionsOur data suggest different patterns of infections between females and male and the importance of an increased vigilance of sexually transmitted pathogens to reduce the burden on general population and the sequelae or the complications on reproductive organs.

Valutazione del ruolo delle IgM, IgG, IgA sieriche e delle IgA dello sputo nella diagnosi di polmonite comunitaria da Chlamydia pneumoniae: studio italiano policentrico

To evaluate the incidence of Chlamydia pneumoniae as etiologic agent of community acquired pneumonia (CAP), specific IgM and IgG antibodies anti-C. pneumoniae in serum and IgA in both serum and sputum were detected by a new ELISA-like test (EIA CP-IgG, IgA, IgM – Eurospital, Italy).The study was carried out from January 1999 to July 2001 in sixteen Italian Hospitals on a total of 141 patients with clinical signs of CAP. At a primary inspection (time T-0) serum and sputum samples were taken from 115/141 patients, whereas serum was collected from only 100/141 patients after 30 days (time T-30). At T-0 24/115 (20.8%) patients showed serological markers suggesting primary C. pneumoniae infection. In 23/24 patients the overall serological pattern found at T-0 was confirmed at T-30. In 32/115 patients (27.8%) serological markers of C. pneumoniae suggesting secondary infection were found positive and were confirmed 30 days later. Our data support the possible role played by C. pneumoniae as an important etiologic agent of CAP throughout different geographic areas of Italy.The test was suitable for the laboratory diagnosis of Chlamydia pneumoniae infection. In particular, the presence of specific IgA anti-Chlamydia pneumoniae in both serum and sputum revealed to be useful to define different stages and evolution of infection.

Evaluation of penicillin susceptibility in clinical isolates of Streptococcus pneumoniae oxacillin resistant

Antibiotic resistant pneumococci have been reported from all continents. Because of the importance of pneumococci in the aetiology of life-threatening diseases, the screening for penicillin resistance with oxacillin disc on all clinically significant isolates is suggested. However, discrepancy between the determination of penicillin resistance by oxacillin disc diffusion and the determination of penicillin minimum inhibitory concentration (MIC) has been reported. On this basis we have examined seven strains of Streptococcus pneumoniae isolated from patients recovered for the exacerbation of chronic bronchitis which were oxacillin-resistant. The assay of penicillin MICs showed that three isolates were moderately resistant to this agent, while four isolates resulted sensitive to penicillin as well as to cefotaxime. These results suggest that a further evaluation of penicillin MIC should be performed on those strains of S. pneumoniae resulting oxacillin-resistant.