Biomedical subjects
Publications and source records attributed to Richard A Venezia.
The prevalence of multidrug-resistant bacteria in the community is increasing, and companion animals serve as a potential reservoir for such bacteria. This report describes a case of a companion dog that was treated with multiple courses of antibiotics for a chronic illness and transmitted multidrug-resistant bacteria to a human through a bite.
OBJECTIVES: To evaluate the impact of the rapid differentiation of Staphylococcus aureus from coagulase-negative staphylococci (CoNS) in blood cultures using peptide nucleic acid fluorescence in situ hybridization (PNA FISH) on vancomycin usage, length of patient hospital stay and hospital costs. DESIGN: This was a retrospective, cost-effective analysis of PNA FISH in its initial 3 month implementation period in 2004 in a 650 bed academic medical centre. Blood cultures with Gram-positive cocci in clusters (GPCC) that were negative for S. aureus using the PNA FISH assay were compared with an untested control group in the same period that had similar illness severity and location. We evaluated the effectiveness of the early identification of CoNS by ruling out S. aureus in conjunction with an antimicrobial team (AMT) on antimicrobial therapy, patient length of stay and hospital costs. RESULTS: A total of 139 blood cultures positive with GPCC had PNA FISH results while 84 in the control group did not. Evaluable criteria were met in 53 patients in the PNA FISH group and 34 in the control group. When comparing the results obtained from using the PNA FISH assay with those for the control group, there was a significant reduction in median length of hospital stay from 6 to 4 days (P < 0.05, CI 0.95-1.87) and a trend towards less vancomycin usage with a decrease in associated hospital costs of approximately Dollars 4000 per patient. CONCLUSIONS: The PNA FISH assay is rapid, accurate and reliable and in association with an AMT could decrease hospital length of stay in patients with CoNS bacteraemia in non-intensive care unit settings and prevent excessive vancomycin usage.
BACKGROUND: No simple, cost-effective methods exist to identify patients at high risk for methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci colonization outside intensive care settings. Without such methods, colonized patients are entering hospitals undetected and transmitting these bacteria to other patients. We aimed to develop a highly sensitive, simple-to-administer prediction rule to identify subpopulations of patients at high risk for colonization on hospital admission. METHODS: We conducted a prospective cohort study of adult patients admitted to the general medical and surgical wards of a tertiary-care facility. Data were collected using electronic medical records and an investigator-administered questionnaire. Cultures of anterior nares and the perirectal area were also collected within 48 hours of admission. RESULTS: Among 699 patients who enrolled in this study, 697 underwent nasal cultures; 555, perirectal cultures; and 553, both. Patient self-report of a hospital admission in the previous year was the most sensitive variable in identifying patients colonized with methicillin-resistant Staphylococcus aureus or with either organism (sensitivity, 76% and 90%, respectively). A prediction rule requiring patients to self-report having received antibiotics and a hospital admission in the previous year would have identified 100% of patients colonized with vancomycin-resistant enterococci. In the high-risk groups defined by the prediction rule, the prevalence of colonization by methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, or either organism were 8.1%, 10.2%, and 15.0%, respectively. CONCLUSION: Patients with a self-reported previous admission within 1 year may represent a high-risk group for colonization by methicillin-resistant Staphylococcus aureus or vancomycin-resistant enterococci at hospital admission and should be considered for targeted active surveillance culturing.
Six cases of Candida dubliniensis fungemia were identified during an 8-month period in hospitalized patients with various conditions, including human immunodeficiency virus infection. Peptide nucleic acid fluorescent in situ hybridization analysis was used as a rapid and reliable test for differentiating C. dubliniensis from Candida albicans, making it feasible to determine the prevalence of C. dubliniensis fungemia.
BACKGROUND: In 1995, beta-lactam inhibitor combinations replaced third-generation cephalosporins as empirical therapy in an effort to manage extended-spectrum beta-lactamase (ESBL) resistance. This study investigated the relationship between antibiotic usage and ESBL organisms from 1994 through 2002 using epidemiological and molecular analysis. METHODS: A case-control study of 119 patients with ESBL organisms and 132 patients with non-ESBL organisms was conducted. Demographics, co-morbidities, device utilization and antibiotic use were analysed for all patients and infected patients only (cases = 75, controls = 83). Both exposure and degree of exposure (in grams) to antibiotics were included. A dot blot hybridization technique was used to identify genes in plasmid extracts from the ESBL organisms. RESULTS: Ventilator days OR 1.1 (1.06, 1.15) P < 0.001, adult respiratory distress syndrome (ARDS) OR 3.1 (1.0, 9.7) P = 0.05, prior aminoglycoside use OR 2.7 (1.2, 6.1) P = 0.02, prior third-generation cephalosporin use OR 7.2 (2.6, 20) P < 0.001, and prior trimethoprim/sulfamethoxazole use OR 8.8 (3.1, 26) P < 0.001 were significantly associated with ESBL organisms by multivariate analysis. All models were concordant with a significant association of ventilator days, third-generation cephalosporins and trimethoprim/sulfamethoxazole with ESBL organisms. beta-Lactamase inhibitor combinations were not associated with ESBL organisms. Hybridization of plasmid extracts demonstrated that 95% of the ESBL organisms carried intI1, a mobile DNA element with a sulphonamide-resistance (R) gene and a frequent carrier of other R factors. Genes for specific types of trimethoprim-R and aminoglycoside-R were present in 26% and 40% of the extracts, respectively. CONCLUSIONS: These data indicate that, besides patient risk factors and third-generation cephalosporins, other antibiotics may provide selective pressures in maintaining ESBL organisms due to multiple resistance genes on plasmids. beta-Lactamase inhibitor combinations appear to be an acceptable substitute to third-generation cephalosporins in strategies to control ESBL organisms.
BACKGROUND: In 2003, the Society of Healthcare Epidemiology of America (SHEA) recommended surveillance cultures upon hospital admission for patients at high risk for carriage of vancomycin-resistant enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA). The aim of this study was to assess the validity of factors from past medical history in defining patients at high risk for subsequent positive cultures with VRE or MRSA upon hospital admission. METHODS: Subjects were adult inpatients admitted to nonintensive care wards of the index hospital during 2001-2002. Cases had MRSA or VRE positive clinical cultures within 48 hours of hospital admission. Patients with previous history of MRSA or VRE were excluded. RESULTS: Nineteen thousand three hundred ninety-nine patients were included, with 273 cases of VRE or MRSA. Previous admission within 1 year of current admission had a sensitivity of 56.8% and a specificity of 88.4% for predicting a case of MRSA or VRE. Individually, the sensitivity and specificity for admission within the past year were 50.5% and 88.4%, respectively, for MRSA and 76.9% and 88.4%, respectively, for VRE. CONCLUSIONS: Patients with a previous hospital admission represent a high-risk population for positive culture for VRE and MRSA and may be a group of which active surveillance is indicated.
The SHV-5 extended-spectrum beta-lactamase gene of pACM1 was previously shown to reside on a segment of DNA ( approximately 7.9 kb) homologous to part of the Klebsiella pneumoniae chromosome. Regions of pACM1 overlapping the ends of the homology were sequenced. A defective copy of IS26 was found on each side of, and immediately adjacent to, the homology. The copies were oriented as direct repeats reminiscent of the compound transposon Tn2680. Other mobile elements and a putative mutagenesis gene, several of which were also defective, were also located in the vicinity of the homology. An intact precursor to the transposon remnant might have contributed to the dissemination of the SHV-5 gene.
Ureaplasma, spp. Mycoplasma genitalium, and Mycoplasma hominis are associated with infection of the genitourinary tract, reproductive failure, and neonatal morbidity and mortality. We have developed a multiplex PCR for the detection of these Mycoplasmatales in a single amplification reaction. The analytical sensitivities of this assay were 10.8, 10.8, and 8.8 CFU for each organism, respectively. This multiplex PCR was compared to culture on 26 cervical swabs, 2 vaginal swabs, 4 female urine specimens, 49 semen samples, 2 male urine specimens, and 1 nonspecified sample. A total of 21 specimens were culture positive (25%); 17 of these were PCR positive. An additional 11 specimens were PCR positive but culture negative. Of the 21 culture-positive specimens, 17 (81%) grew Ureaplasma spp. and 4 (19%) grew Mycoplasma spp. Of the 28 PCR-positive specimens, Ureaplasma spp. was detected in 23 (82%), M. hominis was detected in 3 (11%), and both were detected in 2 (7%). In a confirmatory analysis, all samples were tested by amplification of a second target of the ureaplasma genome. True-positive cases were defined as a positive result by culture or by both amplification assays. The multiplex PCR detected organisms in 26 of the 30 true-positive specimens, as well as in 2 other specimens. Based on a 36% prevalence of infection, the sensitivity, specificity, and positive and negative predictive values of multiplex PCR analyses were 87, 96, 94, and 93%, respectively. Multiplex PCR offers a rapid, sensitive, and easy method to detect genital mycoplasmas.
BACKGROUND: Defining risk factors for central venous catheter (CVC)-associated bloodstream infections (BSIs) is critical to establishing prevention measures, especially for factors such as nurse staffing and antimicrobial-impregnated CVCs. METHODS: We prospectively monitored CVCs, nurse staffing, and patient-related variables for CVC-associated BSIs among adults admitted to eight ICUs during 2 years. RESULTS: A total of 240 CVC-associated BSIs (2.8%) were identified among 4,535 patients, representing 8,593 CVCs. Antimicrobial-impregnated CVCs reduced the risk for CVC-associated BSI only among patients whose CVC was used to administer total parenteral nutrition (TPN, 2.6 CVC-associated BSIs per 1,000 CVC-days vs no TPN, 7.5 CVC-associated BSIs per 1,000 CVC-days; P = .006). Among patients not receiving TPN, there was an increase in the risk of CVC-associated BSI in patients cared for by "float" nurses for more than 60% of the duration of the CVC. In multivariable analysis, risk factors for CVC-associated BSIs were the use of TPN in non-antimicrobial-impregnated CVCs (P = .0001), patient cared for by a float nurse for more than 60% of CVC-days (P = .0019), no antibiotics administered to the patient within 48 hours of insertion (P = .0001), and patient unarousable for 70% or more of the duration of the CVC (P = .0001). Peripherally inserted central catheters (PICCs) were associated with a lower risk for CVC-associated BSI (P = .0001). CONCLUSIONS: Antimicrobial-impregnated CVCs reduced the risk of CVC-associated BSI by 66% in patients receiving TPN. Limiting the use of float nurses for ICU patients with CVCs and the use of PICCs may also reduce the risk of CVC-associated BSI.
Clinical isolates of Enterobacteriaceae with reduced susceptibilities to cephalosporins were collected from 1993 to 2000. The organisms were screened for the extended-spectrum beta-lactamase (ESBL) phenotype, and plasmid extracts were screened for genetic markers by hybridization. A bla(TEM) probe was derived from pUC19; other probes were derived from pACM1, the plasmid responsible for the first known appearance of an ESBL in our institution. These probes included bla(SHV), int, aac(3)-Ia, dfrA1, IS6100, tetA, IncM markers, and Anon 13, a marker for the Klebsiella pneumoniae chromosomal sequences that flank bla(SHV-5). There were 42 hybridization patterns among 237 isolates. Patterns designated pACM1-like occurred in 44% of the isolates (eight species) and were always associated with the clavulanic acid (CA)-susceptible ESBL phenotype. The TEM marker was not predictive of the ESBL phenotype. Mapping indicated the presence of an SHV marker and up to 7.5 kb of its flanking chromosomal sequences in three non-IncM plasmids obtained in transformation experiments. We theorize that this DNA segment spread to other plasmids from pACM1-like sources. CA insensitivity became more frequent with time and was usually associated with either the TEM marker or the absence of both bla markers. One plasmid-encoded enzyme with characteristics of an AmpC beta-lactamase was observed in a transformant lacking both TEM and SHV markers. Although SHV type ESBLs were a continuing source of reduced susceptibility to cephalosporins in our institution, organisms with different resistance mechanisms were added to the hospital microflora in later years. These changes might be related, in part, to ESBL control strategies implemented in 1995.
The nucleotide sequence was determined for Anon 13, a 1250-bp SmaI fragment located approximately 2.8 kb downstream from bla(SHV-5) in pACM1. Anon 13 is 99% identical to a segment of the unpublished sequence of the Klebsiella pneumoniae chromosome. Genes of the K. pneumoniae sequence are undefined, but conceptual amino acid translations of two ORFs in Anon 13 are homologous to L-fuculose-1-phosphate aldolase (FucA) and a conserved hypothetical protein present in the chromosomes of several species of bacteria. In addition, restriction mapping indicates that the region of homology between the K. pneumoniae chromosome and pACM1 is as least 7.9 kb and includes both Anon 13 and bla(SHV). These observations demonstrate the chromosomal origin of the bla(SHV-5) on pACM1.
Methicillin-resistant Staphylococcus aureus (MRSA) is a major nosocomial pathogen worldwide. To investigate an association between antimicrobial use and MRSA, a case control study of 121 patients infected with MRSA compared with 123 patients infected with methicillin-susceptible S. aureus (MSSA) was carried out. Antimicrobial use was analysed by three different logistic regression models: all beta-lactam antibiotics, beta-lactam antibiotics grouped in classes and antimicrobial use in grammes. Patients infected with MRSA tended to have more co-morbidities, longer lengths of stay (LOS) and greater exposure to antibiotics than MSSA-infected patients. Multivariate analysis identified levofloxacin [odds ratio (OR) 8.01], macrolides (OR 4.06), previous hospitalization (OR 1.95), enteral feedings (OR 2.55), surgery (OR 2.24) and LOS before culture (OR 1.03) as independently associated with MRSA infection. All models were concordant with the exception of macrolides, which were not significant based on the number of grammes administered. There were no significant differences in the types of infection or the attributed mortality in either group. MRSA-infected patients had a significantly longer LOS before infection [18.8 +/- 18.2 compared with 8.4 +/- 6.9 (P < 0.001)] and a significantly longer post-diagnosis LOS [27.8 +/- 32.9 compared with 18.6 +/- 21 (P = 0.01)] than MSSA-infected patients.
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