Acinetobacter baumannii has become a major nosocomial threat, causing infections with high rates of morbidity and mortality (1-4).Acinetobacter baumannii hospital outbreaks have been assessed with various DNA typing methods (5-8). Very few typing methods assess outbreaks in real time. Recently, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been developed for the rapid identification of pathogens (9). There are a growing number of reports showing the utility of MALDI-TOF MS to type different bacterial species for easy and early detection of nosocomial outbreaks (10-16).The aim of this study was to assess the potential utility of MALDI-TOF MS to detect the nosocomial spread of A. baumannii isolates. For this purpose, 35 multidrug-resistant strains of A. baumannii, isolated from colonized or infected patients hospitalized in a general hospital in Italy in 2010, were studied using the repetitive sequence-based PCR (rep-PCR) DiversiLab system (DL; bioMérieux, Marcy l'Etoile, France) and MALDI-TOF MS system (Bruker Daltonics, Bremen, Germany), and the results were compared. After storage at Ϫ70°C, the isolates were thawed out and cultured simultaneously on MacConkey agar.DiversiLab rep-PCR (bioMérieux) was performed according to the manufacturer's instructions (5, 17). Briefly, DNA was extracted from a bacterial culture, amplified using rep-PCR, loaded in LabChip, and run using an Agilent 2100 Bioanalyzer (Agilent Technologies). The results were analyzed by DiversiLab with the Pearson correlation (PC) coefficient to emphasize peak intensities more than peak presence or absence. The strains were considered indistinguishable, similar, or different if the similarity was Ն97.5% without differences in the fingerprinting pattern, Ͼ95% and Ͻ97.5%, or Յ95%, respectively.For MALDI-TOF MS, bacteria were cultured for 24 h at 37°C on MacConkey agar and extracts were prepared as described previously (18, 19). For isolate identification, the row spectra were compared with those in the Biotyper database and a log(score) of Ն2.3 was considered to represent a secure species identification. Spectra were acquired with a microflex LT mass spectrometer (Bruker Daltonics) and recorded in the positive linear mode at a laser frequency of 20 Hz, ion source 1 voltage of 20 kV, ion source 2 voltage of 8.5 kV, and mass range from 2,000 to 20,000 kDa, as described elsewhere (18). Reference spectra of newly created main spectra were added to the original Biotyper database. To evaluate the spectrum variation within each strain and to estimate the mass spectral variance of biological replicates, a principal component analysis (PCA) was performed; two biological replicates from six randomly selected Acinetobacter strains were tested, as described above, and spectra for each selected strain were acquired in the same experiment on the mass spectrometer. Technical mass-spectral variance was also evaluated, analyzing five technical replicates from each Acinetobacter independent culture on the same MALDI t...

The identification of reliable molecular biomarkers in prostate cancer early diagnosis is clinically desirable. We quantitatively detected prostate cancer specifically overexpressed genes, DD3 and PSA, in urine sediments of men suffering from prostate cancer or benign prostate hyperplasia, after prostatic massage. As both genes are exclusively expressed in androgen receptor expressing human prostate carcinoma cell lines, we further investigated the possible effect of androgens on PSA and DD3 gene expression. DD3 and PSA mRNA levels were measured by real-time polymerase chain reaction. The combined markers test had a sensitivity of 80.2% and a specificity of 100%. Both gene transcripts were significantly upregulated by androgens. Results indicated the clinical usefulness of the combination of DD3 and PSA as molecular markers in the early diagnosis of prostate cancer and the need of combining as many as possible analytical data with the clinical and demographic ones to achieve the maximum level of diagnostic accuracy.

In this study we report the analysis of 131 Klebsiella pneumoniae (K. pneumoniae) clinical isolates from patients hospitalized in various wards, of Perugia General Hospital, from August 2014 to January 2015. Forty two isolates (32.1 %), were resistant to at least one carbapenem antibiotic and, among these isolates, 14 (33.3 %) exhibited resistance to colistin. All isolates were carbapenemases producers and 41 (97.6 %) harboured the bla KPC gene. Carbapenem-resistant K. pneumoniae isolates (CRKPs) were, also, typed for the genotypic diversity and the results revealed the circulation of two major clusters.This surveillance study evidences the spread of CRKP isolates in Perugia General Hospital and confirms that carbapenem-resistant K. pneumoniae isolates have reached epidemic dissemination in Italy. In addition the percentage of resistance to colistin resulted to be less than that observed in other hospital laboratories across Italy. In conclusion the circulation of these isolates should be monitored and appropriate policy of surveillance must be used, in a target manner, in order to reduce the spread of carbapenem-resistant isolates.