The purpose of this study was to identify carbapenem-resistant Klebsiella pneumoniae in a tertiary care hospital in Sharjah Emirate, to identify the plasmids carrying the carbapenemase genes and to reveal clonal relationships among the isolates. Two hundred and two clinically relevant isolates collected between September 2011 and October 2012 at Al-Qassimi hospital, Sharjah, were investigated for meropenem resistance. Strains with decreased susceptibility were further tested with the modified Hodge test, by EDTA and phenylboronic acid synergy and by E-test. The genes of New Delhi metallo-β-lactamase (NDM), IMP, VIM, OXA-48, and KPC beta-lactamases were targeted by polymerase chain reaction and the genes were located on plasmids by Southern blotting. Clusters of the isolates were revealed by macrorestriction analysis. Seven percent of the isolates were originally found to be meropenem resistant, one isolate have lost its resistance during storage. All of the 13 resistant isolates were positive for the NDM-1 gene located on plasmids of 125 to >170 kb, while three isolates also carried the blaOXA-48-like genes. Clusters having repeatedly been isolated over the study period were identified. Carbapenem-resistant Klebsiella pneumoniae carrying the blaNDM-1 gene is a fast emerging problem, emphasizing the potential role of the Middle East as a secondary reservoir for these organisms.

Background: Typhoid infection remains a major cause of global morbidity. Effective vaccination programmes and new diagnostic tests are urgently needed but are hindered by incomplete understanding of S.Typhi pathogenesis, in part due to insufficiently sensitive methods for detecting bacteria in the peripheral circulation of those encountering infection. Here, new insights into S.Typhi pathogenesis gained using a culture-PCR methodology during a human typhoid challenge model are described.Methods: 40 healthy adult participants were challenged with S.Typhi (Quailes strain) at doses of 1-5 × 103 or 1-5 × 104 colonyforming units. During the 2-weeks after challenge, participants were reviewed daily with clinical data and specimen collection, including blood drawn for 'routine' microbiological culture and a novel culture-PCR assay. For this, 5 mL venous whole-blood was collected into heparin prior to 5-hour culture in 5% ox-bile tryptone soya broth. Centrifugation was performed to collect the blood pellet/bacteria and DNA was extracted using a commercial bloodspin kit. PCR using primers to amplify the fliC-d gene was performed.Results: Bacterial DNA was detected in the peripheral circulation in 57/684 (8.3%) culture-PCR and 53/674 (7.9%) routine blood culture samples. Positive culture-PCR results were detected from 12 hours after oral challenge; 10/40 participants had positive culture-PCR results (but negative routine cultures) within 5 days of challenge. Seven of these participants went on to develop typhoid infection during the 2-week challenge period (typhoid diagnosis defined by development of bacteraemia or persistent fever >38 • C for >12-hours). DNA was detected in the peripheral circulation of 5/40 participants who were not diagnosed with typhoid infection during the challenge period. Several of these participants had mild symptoms or elevation of inflammatory markers (including C-reactive protein) only.Conclusion: These data suggest that a culture-PCR methodology targeting the fliC-d gene may be used to detect DNA in peripheral blood of those challenged withS.Typhi. Aside from unique confirmation that the mechanism of typhoid infection includes primary dissemination of bacteria in the peripheral circulation, we also demonstrate that asymptomatic infection/circulation of bacteria maybe more common than previously anticipated. Sensitive detection of S.Typhi DNA in peripheral blood samples may represent a useful additional endpoint in the evaluation of typhoid vaccines.

Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) is an emerging pathogen in hospitalized patients worldwide. The present study was undertaken to identify CA-MRSA in hospitalized patients in a 350-bed tertiary care hospital in Sharjah, UAE over a 2-year period from January 2011 to December 2012. CA-MRSA was defined based on identification within first 48 h of admission in the hospital. Staphylococcal cassette chromosome (SCC) mec typing of the CA-MRSA isolates was carried out by multiplex polymerase chain reaction (PCR). Detection of PVL and mecA genes was done by PCR using the GenoType(®) MRSA test system (Hain Lifescience). Patient's clinical data and antimicrobial susceptibility pattern of the CA-MRSA isolates were also evaluated. Fifty seven of the 187 MRSA isolates were identified as CA-MRSA. All the CA-MRSA strains in our study belonged to SCCmecIV type and were positive for both PVL and mecA genes. The patients with CA-MRSA infections were young (median age, 32 years) and the majority of infections involved the skin and soft tissue (36%). Antimicrobial susceptibility pattern of the CA-MRSA isolates showed a better susceptibility profile to the non-beta-lactam antimicrobials with the exception of ciprofloxacin having 28% resistance. This study evidently strengthens the recent observation of an increase in CA-MRSA emergence among hospitalized patients in the UAE.