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      Usefulness of 23S rRNA Amplification by PCR in the Detection of Bacteria in CAPD Peritonitis

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          Background: Peritonitis is the most common complication of continuous ambulatory peritoneal dialysis (CAPD), and the spectrum of organisms causing CAPD peritonitis is broad. Polymerase chain reaction (PCR) has recently broadened its diagnostic capabilities in infectious diseases. PCR can provide a sensitive method for identifying causative infectious organisms. Methods: To evaluate the usefulness of 23S bacterial ribosomal RNA amplification and direct sequencing for the detection of infectious organisms, we compared PCR with bacteriological culture for the analysis of dialysates from CAPD peritonitis patients. Thirty-two samples from CAPD peritonitis patients with current antibiotic use and control samples from 30 CAPD patients without peritonitis were examined by PCR with sequencing analysis and by conventional bacteriological culture. In addition, 95 culture-positive samples and 39 culture-negative samples from CAPD peritonitis patients before antibiotic treatment were analyzed by PCR assay. Results: In the control samples from patients without CAPD peritonitis, false-positive rates were relatively rare: 3 of 30 in the PCR study and 2 of 30 in the culture study. Of the 134 CAPD peritonitis samples collected before antibiotic therapy, positive cultures were obtained in 70.9% (95/134) of them. In 75 of the culture-positive samples, the same microorganisms were confirmed by PCR assay, and the others showed discrepant results as compared with culture study. In 30 of the 39 culture-negative samples, microbial organisms were detected by PCR assay. Of the 32 samples from patients who developed CAPD peritonitis during antibiotic treatment, 17 (53.1%) were positive by PCR assay, and 5 (15.6%) were positive by culture. Conclusion: Our study suggests that broad-spectrum PCR with RNA sequencing can complement culture methods in the diagnosis of CAPD peritonitis, especially in patients with previous or current antibiotic use.

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          Most cited references 7

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          Carriage of methicillin-resistant Staphylococcus aureus among hospital employees: prevalence, duration, and transmission to households.

          To assess the prevalence and duration of methicillin-resistant Staphylococcus aureus (MRSA) carriage among hospital employees and transmission to their households. A point-prevalence survey of MRSA carriage (nasal swabbing) of staff and patients throughout the hospital; a prevalence survey of MRSA carriage in 2 medical wards, with carriers observed to estimate carriage duration; and evaluation of transmission to MRSA-positive workers' families. All MRSA isolates were analyzed by pulsed-field gel electrophoresis. During the study, no MRSA outbreak was detected among hospitalized patients. A 600-bed, public tertiary-care teaching hospital near Paris. Sixty MRSA carriers were identified among 965 healthcare providers (prevalence, 6.2%; CI95, 4.7%-7.7%). Prevalence was higher in staff from clinical wards than from elsewhere (9.0% vs 2.1%; P < .0001). Identity of isolates from employees and patients varied from 25% in medical wards to 100% in the long-term-care facility. MRSA carriage was identified in 14 employees from 2 medical wards (prevalence, 19.4%; CI95, 10.3%-28.5%). Prevalence depended on the length of service in these wards. Transmission to households was investigated in 10 MRSA-positive workers' families and was found in 4. All isolates from each family were identical. Few data are available concerning the prevalence of MRSA carriers among hospital employees in the absence of an outbreak among patients. MRSA transmission between patients and employees likely depends on the frequency and duration of exposure to MRSA-positive patients and infection control measures employed. Frequent transmission of MRSA from colonized healthcare workers to their households was documented.
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            Analysis of microbiological trends in peritoneal dialysis-related peritonitis from 1991 to 1998.

            The microbial cause of peritoneal dialysis-related peritonitis is an important determinant of clinical outcome and the basis of widely used treatment guidelines. Five hundred forty-six cases of peritonitis in 374 patients from 1991 to 1998 were analyzed. The rate of peritonitis declined significantly from 1.37 episodes/patient-year in 1991 to 0.55 episode/patient-year in 1998 (P = 0.02). The rate of Gram-positive peritonitis decreased significantly from 0.75 to 0.28 episode/patient-year during the same period (P = 0.02). Conversely, the occurrence of Gram-negative peritonitis remained constant at approximately 0.16 episode/patient-year (P = 0.28). Staphylococcus epidermidis and Staphylococcus aureus were the most common causes of peritonitis, isolated in 27.8% and 19.3% of the culture-positive cases, respectively. A distinct decrease in peritonitis caused by S epidermidis was observed, with 0.40 episode/patient-year in 1991 compared with 0.11 to 0.20 episode/patient-year during subsequent years. The rate of infections caused by S aureus decreased significantly over time from a high of 0.21 episode/patient-year in 1992 to a low of 0.04 episode/patient-year in 1998 (P = 0.01). Pseudomonas aeruginosa, Escherichia coli, and KLEBSIELLA: species were the most common causes of Gram-negative peritonitis, identified in 7.1%, 6.8%, and 5.2% of culture-positive cases, respectively. The most dramatic increase in antibiotic resistance was seen among S epidermidis. From 1991 and 1992 to 1997 and 1998, resistance to ciprofloxacin increased from 5.4% to 47.8% (P = 0.003), and resistance to methicillin increased from 18.9% to 73.9% (P = 0.03). Our study showed significant trends in the causative pathogens of peritoneal dialysis-related peritonitis and dramatic increases in antibiotic resistance. These data support further study and warrant reevaluation of current treatment practices.
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              Rapid identification of yeasts in positive blood cultures by a multiplex PCR method.

              Yeasts are emerging as important etiological agents of nosocomial bloodstream infections. A multiplex PCR method was developed to rapidly identify clinically important yeasts that cause fungemia. The method amplified the internal transcribed spacer 1 (ITS1) region between the 18S and 5.8S rRNA genes and a specific DNA fragment within the ITS2 region of Candida albicans. With this method, C. albicans produced two amplicons, whereas other species produced only one. Through sequence analysis, the precise lengths of the PCR products were found to be as follows: C. glabrata (482 or 483 bp), C. guilliermondii (248 bp), C. parapsilosis (229 bp), C. albicans (218 or 219 and 110 bp), C. tropicalis (218 bp), Cryptococcus neoformans (201 bp), and C. krusei (182 bp). The PCR products could be effectively separated by disk polyacrylamide gel electrophoresis. The method was used to test 249 positive blood cultures (255 isolates), from which the following species (strain number) were isolated: C. albicans (128), C. tropicalis (51), C. glabrata (28), C. parapsilosis (23), C. neoformans (9), C. krusei (5), C. guilliermondii (3), and other, minor species (8). The test sensitivity of the method was 96.9% (247 of 255 isolates). The eight minor species were either misidentified (one strain) or not identified (seven strains). From the time at which a positive bottle was found, the multiplex PCR could be completed within 8 h; the present method is simpler than any previously reported molecular method for the identification of blood yeasts.

                Author and article information

                Am J Nephrol
                American Journal of Nephrology
                S. Karger AG
                May 2006
                02 June 2006
                : 26
                : 2
                : 115-120
                Divisions of aNephrology and bInfection, Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea
                92040 Am J Nephrol 2006;26:115–120
                © 2006 S. Karger AG, Basel

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                Page count
                Tables: 7, References: 19, Pages: 6
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                Original Report: Laboratory Investigation


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