Utility of a pre-optimized kit for random amplified polymorphic DNA in typing Candida albicans.

The utility of a pre-optimized kit for random amplified polymorphic DNA (RAPD) was assessed in typing diverse strains of Candida albicans from epidemiologically unrelated inpatients (interpatient analysis) and in detecting clonal variations that maybe present within individual patient isolates (intrapatient analysis). Stool samples from inpatients were cultured on Inhibitory Mold agar. Nine individual colonies from all patients with > or =9 colonies of C. albicans (n = 18) were selected, frozen, and karyotyped using CHEF genomic DNA plug kits and CHEF-DRIII. Each of the selected colonies was then analyzed by RAPD, utilizing the selected kit, with 6 primers. Interpatient analysis revealed 9 karyotypes and 17 RAPD composites. RAPD discrimination was significantly better (p < 0.001). Intrapatient analysis revealed 34 (21%) and 33 (20.4%) variants among 162 colonies tested by RAPD and karyotyping, respectively. The results were discordant in 25 variants, all with differences of 1-3 bands. These results illustrate that this pre-optimized kit for RAPD provides excellent discrimination of genetically unrelated strains. Its performance in delineating subtle clonal differences was comparable with karyotyping; both methods failed to detect all minor genetic variations. The ease of use and quick turnaround time of this kit offer a practical and reliable method for typing diverse strains of C. albicans, but may be inadequate for assessing microevolution.

Stool colonization with vancomycin-resistant enterococci in healthcare workers and their households.

OBJECTIVE: To determine the prevalence of stool colonization with vancomycin-resistant enterococci (VRE) among healthcare workers (HCWs) and their families. DESIGN: Prospective assessment of fecal colonization with VRE. SETTING: A 603-bed, tertiary-care teaching hospital. PARTICIPANTS: Healthy volunteers recruited from hospital employees and their households were screened to exclude pregnancy, diabetes mellitus, immunosuppressive disorders, and recent use of antimicrobials. INTERVENTION: Self-obtained stool swabs were used to obtain cultures. Isolated enterococci were screened for vancomycin resistance and species were identified. Intra-household isolates were genotyped using pulsed-field gel electrophoresis (PFGE). RESULTS: The participants (n = 228; age range, 28 days to 80 years) were from 137 households with and 91 without employees who had contact with patients. Enterococcus species were isolated from 127 stool specimens (55.7%). VRE were detected in 12 individuals, representing 6 E. casseliflavus, 5 E. faecium, and 1 E. gallinarum. VRE were more commonly isolated in employees who had contact with patients (5 of 52 vs 0 of 40; relative risk [RR], 1.9; 95% confidence interval [CI95], 1.5 to 2.2; P = .07) and their household members (10 of 137 vs 2 of 91; RR, 3.3; CI95, 0.7 to 14.8; P = .13). In 2 households (2 adults in a physician's household and an adult plus a child in a nurse's household) PFGE analysis demonstrated identical intra-household strains of vancomycin-resistant E. faecium. CONCLUSIONS: VRE colonization was found in 5.3% of screened stools and was more prevalent in HCWs who had contact with patients and their households. Identical PFGE patterns between 2 employees who had contact with patients and their household members demonstrated probable intra-household spread. Although the mode of acquisition was uncertain, the association with employees who had contact with patients suggests possible occupational sources. These findings demonstrate the spread of VRE within the household and implicate occupational risk for its acquisition.