Antibodies to hepatitis E virus among several populations in Greece: increased prevalence in an hemodialysis unit

BACKGROUND: Hepatitis E virus (HEV) has been found to be the causative agent of enterically transmitted non-A, non-B hepatitis in tropical and subtropical countries. Several investigators, however, have indicated that HEV could be endemic in Europe, albeit at a low prevalence. STUDY DESIGN AND METHODS: The purpose of this study was to estimate the prevalence of anti-HEV in various populations in northwestern Greece (Epirus region). Healthy blood donors (2636), refugees from southern Albania (350), children (165), injecting drug users (IDUs) (65), multiply transfused patients (62), patients with chronic viral hepatitis (75), and chronic hemodialysis patients (149) were investigated for anti-HEV by enzyme immunoassay and confirmatory Western blot assay. In addition, 380 consecutive healthy blood donors and 62 hemodialysis patients from a neighboring area (Agrinion, Greece) were investigated. RESULTS: A very low presence of anti-HEV antibody was found among healthy blood donors from Epirus (0.23%) and Agrinion (0.53%). Anti-HEV was not detected in children, IDUs, or multiply transfused patients. In contrast, a low but significant prevalence of anti-HEV was found among refugees (4.85%), patients with chronic viral hepatitis (5.3%), and hemodialysis patients from Epirus (1.34%), as compared with healthy blood donors from Epirus: p < 0.0001, p < 0.00001, and p < 0.10, respectively. A high prevalence (9.7%) of anti- HEV was revealed in patients at the hemodialysis unit of the General Hospital of Agrinion (p < 0.00005, compared to healthy blood donors from Agrinion). No significant association was found between anti-HEV positivity and the age or sex of donors, the duration of hemodialysis, positivity for hepatitis B or C virus infection markers, history of hepatitis, increased alanine aminotransferase, renal transplantation, a history of transfusion, or the number of units transfused. CONCLUSION: This study demonstrated a high prevalence of anti-HEV in a separate hemodialysis unit, without an association with the known routes of transmission of blood-borne viruses. This observation suggests that a still-undefined intra-unit factor or other factors are associated with HEV transmission.     

Evaluation of a new line probe assay for rapid identification of gyrA mutations in Mycobacterium tuberculosis

Resistance of Mycobacterium tuberculosis to fluoroquinolones (FQ) results mostly from mutations in the gyrA gene. We developed a reverse hybridization-based line probe assay in which oligonucleotide probes carrying the wild-type gyrA sequence, a serine-to-threonine (S95T) polymorphism, and gyrA mutations (A90V, A90V-S95T, S91P, S91P-S95T, D94A, D94N, D94G-S95T, D94H-S95T) were immobilized on nitrocellulose strips and hybridized with digoxigenin-labeled PCR products obtained from M. tuberculosis strains. When a mutated PCR product was used, hybridization occurred to the corresponding mutated probe but not to the wild-type probe. A panel of M. tuberculosis complex strains including 19 ofloxacin-resistant (OFL-R) and 9 ofloxacin-susceptible (OFL-S) M. tuberculosis strains was studied for detection and identification of gyrA mutations by the line probe assay and nucleotide sequencing, in comparison with testing of in vitro susceptibility to FQ. Results were 100% concordant with those of nucleotide sequencing. The S95T polymorphism, which is not related to FQ resistance, was found in 5 OFL-S and 2 OFL-R strains; the other 17 OFL-R strains harbored single mutations associated with serine or threonine at codon 95. No mutations were found in the other OFL-S strains. Overall, on the basis of the MICs on solid medium, the new line probe assay correctly identified all OFL-S and 17 out of 19 (89.5%) OFL-R strains. A nested-PCR protocol was also evaluated for the assay to amplify PCR products from M. tuberculosis-spiked sputa, with a good specificity and a sensitivity of 2 x 10(3) M. tuberculosis CFU per ml of sputum.     

Renal transplantation done safely without prior chronic dialysis therapy

The complications, cost, and inconvenience associated with pretransplant hemodialysis and peritoneal dialysis would be minimized if transplantation were instituted without prior dialysis. That preuremic transplantation is safe and efficacious in patients with immanent end-stage renal disease has not been established. All 1742 consecutive primary renal transplants performed at the University of Minnesota during the 16.5 year period from January 1968 through July 1984 were reviewed to determine whether graft and patient survival were adversely affected by transplantation prior to dialytic therapy. In the overall group of primary renal transplants, no differences in actuarial graft or patient survival were noted with or without prior dialysis. Likewise, outcome was not affected by the pretransplant dialysis status in recipients of allografts from HLA-identical mismatched living-related donors. However, in cadaveric transplantation graft function appeared to be adversely affected by transplantation prior to dialysis, with 52% vs. 66% two-year graft function for nondialyzed vs. chronically dialyzed recipients, respectively (P = 0.15). Patient survival was significantly (P = .04) decreased in the nondialyzed group, with 66% vs. 80% two-year survival in the chronic dialysis group. However, nearly all of the nondialyzed, cadaveric recipients were diabetic. The outcome of transplantation was found to be identical in these patients, as compared with chronically dialyzed diabetic recipients of cadaveric grafts. Thus, the apparent detrimental effect of predialytic transplantation in the cadaver group was due to the preponderance of diabetics in the nondialyzed group. Since July 1984, a single-armed therapeutic trial of combination therapy with azathioprine, prednisone, antilymphoblast globulin (ALG), and cyclosporine has been undertaken, Since that time, 36 primary graft recipients were transplanted prior to dialysis. Of these 36, 35 currently have a functioning graft. Thus, transplantation prior to chronic dialysis is safe irrespective of donor source, or choice of immunosuppressive agents.