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- CMV infection of liver transplant recipients: comparison of antigenemia and molecular biology assays.Publication . Amorim, M.; Cabeda, J.; Seca, R.; Mendes, A.; Castro, A.; Amorim, J.Abstract BACKGROUND: CMV is a major clinical problem in transplant recipients. Thus, it is important to use sensitive and specific diagnostic techniques to rapidly and accurately detect CMV infection and identify patients at risk of developing CMV disease. In the present study, CMV infection after liver transplantation was monitored retrospectively by two molecular biology assays - a quantitative PCR assay and a qualitative NASBA assay. The results were compared with those obtained by prospective pp65 antigenemia determinations. MATERIALS AND METHODS: 87 consecutive samples from 10 liver transplanted patients were tested for CMV by pp65 antigenemia, and CMV monitor and NASBA pp67 mRNA assay. RESULTS: CMV infection was detected in all patients by antigenemia and CMV monitor, whereas NASBA assay identified only 8/10 patients with viremia. Furthermore, CMV infection was never detected earlier by molecular biology assays than by antigenemia. Only 5/10 patients with CMV infection developed CMV disease. Using a cut off value of 8 cells/50,000, antigenemia was found to be the assay that better identified patients at risk of developing CMV disease. However, the kinetics of the onset of infection detected by NASBA and CMV monitor seemed to have better identified patients at risk of developing CMV disease. Furthermore, before onset of disease, CMV pp67 mRNA was found to have similar or better negative and positive predictive values for the development of CMV disease. CONCLUSIONS: The present data, suggests that the concomitant use of antigenemia and pp67 mRNA assay gives the best identification of patients at risk of developing CMV disease.
- Three years incidence of dermatophytes in a hospital in Porto (Portugal)Publication . LOPES, V.; VELHO, G.; AMORIM, M.L.; CARDOSO, M.L.; MASSA, A.; AMORIM, J.M.Rev Iberoam Micol. 2002 Dec;19(4):201-3. [Three years incidence of dermatophytes in a hospital in Porto (Portugal)]. [Article in Spanish] Lopes V, Velho G, Amorim ML, Cardoso ML, Massa A, Amorim JM. SourceServiço de Microbiologia, Hospital Geral Santo António, Largo Prof. Abel Salazar, 4099-001 Porto, Portugal. Abstract We evaluated the incidence of dermatophytes isolated at our hospital in the years of 1997 to 2000 and correlated it with anatomical site and age. Trichophyton rubrum was the predominant species in all anatomical sites, excluding scalp, followed by Microsporum canis, the leading agent of tinea capitis. All dermatophytosis, except tinea capitis by M. canis and Trichophyton schoenleinnii appeared mainly in adult patients. Our results revealed no substantial differences to other portuguese studies regarding the major agents. We found a relatively high incidence of T. schoenleinnii as second tinea capitis agent.
- Systemic lupus erythematosus in Europe at the change of the millennium:Lessons from the "Euro-Lupus Cohort"Publication . Cervera, R.; Abarca-Costalago, M.; Abramovicz, D.; Allegri, F.; Annunziata, P.; Ayditung, A.; Bacarelli, M.; Bellisai, F.; Bernardino, I.; BIERNAT‐KALUZA, M.; BLOCKMANS, D.; BOKI, K.; BRACCI, L.; Campanella, V.; Camps, M.; Carcassi, C.; Cattaneo, R.; Cauli, A.; Chwalinska‐Sadowska, H.; Contu, I.; Cosyns, J.; Danieli, M.; D'cruz, D.; Depresseux, G.; Direskeneli, H.; Domènech, I.; Fernández‐Nebro, A.; Ferrara, G.; Font, J.; Frutos, M.; Galeazzi, M.; García‐Carrasco, M.; García-Iglesias, M.; García‐Tobaruela, A.; George, J.; Gil, A.; González‐Santos, P.; Grana, M.; Gül, A.; Haga, H.; De Haro‐Liger, M.; Houssiau, F.; Hughes, G.; Ingelmo, M.; Jedryka‐Góral, A.; khamashta, M.; Lavilla, P.; Levi, Y.; López‐Dupla, M.; López‐Soto, A.; Maldykowa, H.; Marcolongo, R.; Mathieu, A.; Morozzi, G.; Nicolopoulou, N.; Papasteriades, C.; Passiu, G.; Perelló, I.; Petera, P.; Petrovic, R.; Piette, J.; Pintado, V.; De Pita, O.; Popovic, R.; Pucci, G.; Puddu, P.; De Ramón, E.; Ramos‐Casals, M.; Rodríguez‐Andreu, J.; Ruiz‐Irastroza, G.; Sánchez‐Lora, J.; Sanna, G.; Scorza, R.; Sebastini, G.; Sherer, Y.; Shoenfeld, Y.; Simpatico, A.; Sinico, R.; Smolen, J.; Tincani, A.; Tokgöz, G.; Urbanomárquez, A.; Vasconcelos, C.; Vázquez, J.; Veronesi, M.; Vianni, J.; Vivancos, J.The bEuro-Lupus CohortQ is composed by 1000 patients with systemic lupus erythematosus (SLE) that have been followed prospectively since 1991. These patients have been gathered by a European consortium—the bEuro-Lupus Project GroupQ. This consortium was originated as part of the network promoted by the bEuropean Working Party on SLEQ, a working group created in 1990 in order to promote research in Europe on the different problems related to this disease. The bEuro-Lupus CohortQ provides an updated information on the SLE morbidity and mortality characteristics in the present decade as well as defines several clinical and immunological prognostic factors
- Changes in the Clonal Nature and Antibiotic Resistance Profiles of Methicillin-Resistant Staphylococcus aureus Isolates Associated with Spread of the EMRSA-15 Clone in a Tertiary Care Portuguese HospitalPublication . Amorim, M.; Faria, N.; Oliveira, D.; Vasconcelos, C.; Cabeda, J.; Mendes, A.; Calado, E.; Castro, A.; Ramos, M.; Amorim, J.; Lencastre, H.Abstract Two hundred eighty methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates recovered from a tertiary care hospital in Oporto, Portugal, between 2003 and 2005 were studied by a combination of molecular typing techniques in order to investigate the genetic backgrounds associated with the changes in the resistance phenotypes observed since 2001 and compare them to those previously found in the hospital. All MRSA isolates were grouped into resistance profiles for a panel of seven antibiotics and characterized by pulsed-field gel electrophoresis (PFGE) and SCCmec (staphylococcal cassette chromosome mec) typing. Representative isolates of PFGE types were further studied by spa typing and multilocus sequence typing. Our findings clearly document that the increasing isolation of nonmultiresistant MRSA strains was associated with the decline (from 69% in 1996 to 2000 to 12% in 2003 to 2005) and massive replacement of the multiresistant Brazilian clone (ST239-IIIA) by the epidemic EMRSA-15 clone (ST22-IV), in which resistance to antibiotics other than beta-lactams is very rare, as the major clone (80% of isolates). The Iberian clone (ST247-IA), a major clone in 1992 to 1993, was represented in the present study by just one isolate. Two other pandemic MRSA clones were detected, as sporadic isolates, for the first time in our hospital: the New York/Japan (ST5-II) and the EMRSA-16 (ST36-II) clones. Furthermore, the pattern of susceptibility of MRSA isolates both to gentamicin and to trimethoprim-sulfamethoxazole was shown to be an excellent phenotypic marker for the discrimination of the EMRSA-15 clone from other nonmultiresistant MRSA clones present in our hospital.
- First trimester aneuploidy screening program for preeclampsia prediction in a portuguese obstetric populationPublication . Teixeira, C.; Tejera, E.; Martins, H.; Pereira, A.; Costa-Pereira, A; Rebelo, IObjective. To evaluate the performance of a first trimester aneuploidy screening program for preeclampsia (PE) prediction in a Portuguese obstetric population, when performed under routine clinical conditions. Materials and Methods. Retrospective cohort study of 5672 pregnant women who underwent routine first trimester aneuploidy screening in a Portuguese university hospital from January 2009 to June 2013. Logistic regression-based predictive models were developed for prediction of PE based on maternal characteristics, crown-rump length (CRL), nuchal translucency thickness (NT), and maternal serum levels of pregnancy-associated plasma protein-A (PAPP-A) and free beta-subunit of human chorionic gonadotropin (free β -hCG). Results. At a false-positive rate of 5/10%, the detection rate for early-onset (EO-PE) and late-onset (LO-PE) PE was 31.4/45.7% and 29.5/35.2%, respectively. Although both forms of PE were associated with decreased PAPP-A, logistic regression analysis revealed significant contributions from maternal factors, free β -hCG, CRL, and NT, but not PAPP-A, for prediction of PE. Conclusion. Our findings support that both clinical forms of EO-PE and LO-PE can be predicted using a combination of maternal history and biomarkers assessed at first trimester aneuploidy screening. However, detection rates were modest, suggesting that models need to be improved with additional markers not included in the current aneuploidy screening programs.
- Importation of Fosfomycin Resistance fosA3 Gene to EuropePublication . Mendes, A.; Rodrigues, C.; Pires, J.; Amorim, J.; Ramos, M.; Novais, Â.; Peixe, L.
- Aspergillus Species and Antifungals Susceptibility in Clinical Setting in the North of Portugal: Cryptic Species and Emerging Azoles Resistance in A. fumigatusPublication . Pinto, E.; Monteiro, C.; Maia, M.; Faria, M.; Lopes, V.; Lameiras, C.; Pinheiro, D.Aspergillus spp. are agents of a broad-spectrum of diseases among humans. Their growing resistance to azoles, the cornerstone in the management of human aspergillosis, is a worrisome problem around the world. Considering lack of data from Portugal on this topic, particularly from the northern region, a retrospective surveillance study was planned to assess frequency of cryptic Aspergillus species and azoles resistance. A total of 227 clinical isolates, mainly from the respiratory tract (92.1%), collected from three hospitals serving a population of about three million people, were studied for their epidemiology and antifungal susceptibility patterns determined by the E.DEF.9.3 protocol of EUCAST. Employing molecular methods, seven Aspergillus complexes were identified; Aspergillus fumigatus sensu stricto was the most frequent isolate (86.7%). A 7.5% prevalence of cryptic species was found; A. welwitschiae (A. niger complex-3.1%) and A. lentulus (A. fumigatus complex-2.2%) were the most frequent. Amongst cryptic species, it was found a percentage of resistance to voriconazole, posaconazole and isavuconazole of 47.1, 82.4, and 100%, respectively. Five A. fumigatus sensu stricto showed pan-azole resistance. Sequencing their cyp51A gene revealed the presence of one isolate with TR46/Y121F/T289A mutation and two isolates with TR34/L98H mutation. This study emphasizes the need to identify strains to the species level and to evaluate their antifungal susceptibility in all human originated Aspergillus spp. isolates, particularly those from invasive aspergillosis.
- Economic Impact of Prosthetic Joint Infection - an Evaluation Within the Portuguese National Health SystemPublication . Sousa, A.; Carvalho, A.; Pereira, C.; Reis, E.; Santos, A.; Abreu, M.; Soares, D.; Fragoso, R.; Ferreira, S.; Reis, M.; Sousa, R.Introduction: Prosthetic infection is a devastating complication of arthroplasty and carries significant economic burden. The objective of this study was to analyze the economic impact of prosthetic hip and knee infection in Portuguese National Health System. Material and Methods: Case-control study carried out from January 2014 to December 2015. The mean costs of primary arthroplasties and prosthetic revision surgeries for non-infectious reasons were compared with the costs of prosthetic infections treated with debridement and preservation of the prosthesis or with two-stage exchange arthroplasty.The reimbursement for these cases was also evaluated and compared with its real costs. Results: A total of 715 primary arthroplasties, 35 aseptic revisions, 16 surgical debridements and 15 revisions for infectious reasons were evaluated. The cost of primary arthroplasties was 3,230€ in the hips and 3,618€ in the knees. The cost of aseptic revision was 6,089€ in the hips and 7,985€ in the knees. In the cases treated with debridement and implant retention the cost was 5,528€ in the hips and 4,009€ in the knees. In cases of infections treated with a two-stage revision the cost was 11,415€ and 13,793€ for hips and knees, respectively. Conclusion: As far as we know this is the first study that analyzes the economic impact of prosthetic infection in the Portuguese context. Although direct compensation for treating infected cases is much lower than calculated costs, infected cases push the overall hospital case-mix-index upwards thus increasing financial compensation for the entire cohort of treated patients. This knowledge will allow for more informed decisions about health policies in the future.
- Ultra-rapid flow cytometry assay for colistin MIC determination in Enterobacterales, Pseudomonas aeruginosa and Acinetobacter baumanniiPublication . Fonseca e Silva, Daniela; Andrade, Ferdinando F.; Gomes, Rosário; Silva-Dias, Ana; Martins-Oliveira, Inês; Pérez-Viso, Blanca; Ramos, Maria Helena; Rodrigues, Acácio G.; Cantón, Rafael; Pina-Vaz, CidáliaObjectives: Both EUCAST and CLSI recommend broth microdilution for antimicrobial susceptibility testing of colistin, but this method is cumbersome and takes 16-24 h to give results. Our objective was to evaluate a rapid quantitative colistin MIC susceptibility assay based on flow cytometry analysis (FASTcolistin MIC) in comparison with standard broth microdilution assay. Methods: One hundred and sixteen Gram-negative bacilli (78 Enterobacterales, 28 Pseudomonas aeruginosa and 10 Acinetobacter baumannii) were studied in parallel using standard broth microdilution following EUCAST recommendations and FASTcolistin MIC kit. In the last one, a bacteria suspension (0.5 MacFarland) was prepared, diluted in Muller-Hinton broth, incubated in the susceptibility panel containing different colistin concentrations (range 0.125-64 mg/L) with a fluorescent probe and incubated 1 h at 35ºC. After that, a flow cytometry analysis using CytoFLEX (Beckmam) was performed. Using a dedicated software (BioFAST) an automated MIC result was obtained after 1.5 h. Performance evaluation was performed according to the ISO standard 20776-2. Reproducibility and repeatability, categorical (CA) and essential agreement (EA), and lot-to-lot variation and operator-to-operator variability, as well as time to results were determined. Results: Overall, 100% CA (CI 97-100%) and 95.7% EA (CI 90-98%) was obtained with high repeatability (100%; CI 80-100%)and reproducibility (97%; (CI 83-99%)). Absence of lot-to-lot variations or differences in the operators' performance was observed. Conclusions: FASTcolistin MIC is an accurate, reliable and ultra-rapid method (1 h incubation versus 24 h) for susceptibility testing of colistin of common Gram-negative bacilli recovered in clinical laboratories.
- Donor-derived fatal hyperinfection strongyloidiasis in renal transplant recipientPublication . Cipriano, Ana; Dias, Rita; Cleto Marinho, Ricardo; Correia, Sofia; Lopes, Virgínia; Cardoso, Teresa; Aragão, IreneStrongyloides stercoralis is a nematode, endemic in tropical and subtropical areas. Strongyloidiasis has been reported in recipients of hematopoietic stem cells, kidney, liver, heart, intestine, and pancreas, eventually presenting as disseminated strongyloidiasis and hyperinfection syndrome (SHS) which is associated with high mortality. We report one case of a recent renal transplant recipient, who presented with gastrointestinal and respiratory symptoms, evolving into shock. The identification of Strongyloides stercoralis in the bronchoalveolar lavage (BAL) lead us to the diagnosis of SHS. Treatment with subcutaneous ivermectin was started, however the patient did not survive. Retrospective serum donor analysis allowed us to identify the donor as the source of infection.