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Biomedical subjects

D Olden

Publications and source records attributed to D Olden.

8 recordsLinked to original sources

Effect of anti-tuberculosis treatment on the tuberculin interferon-gamma response in tuberculin skin test (TST) positive health care workers and patients with tuberculosis.

SETTING: Public hospital, Victoria, Australia. OBJECTIVE: To evaluate the effect of multidrug treatment and isoniazid (INH) chemoprophylaxis on the tuberculin interferon-y assay (QIFN) in 19 patients with culture-confirmed Mycobacterium tuberculosis and 119 health care workers (HCWs) with tuberculin skin tests (TST) > or =15 mm. DESIGN: Patients with M. tuberculosis were treated with standard medication and tested with QIFN at diagnosis and at regular intervals over a 12-month period. All HCWs, 59 (50%) of whom were prescribed INH chemoprophylaxis, were tested with QIFN at baseline, 2, 4, 6 and 12 months. RESULTS: QIFN results in patients with tuberculosis were consistent and reproducible. At the initial time point QIFN assays were positive for M. tuberculosis in 67%, and once positive, the QIFN assay remained so over the 12-month period. In the HCWS, initial QIFN assays were positive in 73 (61%). During the 12-month study, 91 HCWs had a QIFN assay on at least two occasions. The overall reproducibility between tests was fair (kappa statistic = 0.45), and was little affected by administration of INH. CONCLUSION: These data suggest that although the QIFN assay is generally positive in patients with proven tuberculosis, it does not provide clinically useful information during the first 12 months of treatment with multidrug chemotherapy or INH chemoprophylaxis.

Adult↗

Outcome of a screening program for vancomycin-resistant enterococci in a hospital in Victoria.

OBJECTIVE: To screen for faecal colonisation with vancomycin-resistant enterococci (VRE) among potentially at-risk patients. DESIGN: Infection control screening program. SETTING: Monash Medical Centre (a tertiary care hospital), Melbourne, Victoria, in the seven months from June 1997. PATIENTS: Patients in the Renal, Oncology and Intensive Care (ICU) Units. MAIN OUTCOME MEASURES: Presence of VRE in a rectal swab or faecal specimen taken at admission and at regular intervals during inpatient stay; presence of vancomycin-resistance genes (vanA, vanB and vanC) assessed by polymerase chain reaction (PCR); genetic clonality of isolates assessed by pulsed-field gel electrophoresis (PFGE). RESULTS: 574 patients (356 renal, 134 ICU and 84 oncology) were screened; 12 were colonised with VRE--nine renal inpatients, two having peritoneal dialysis or incentre haemodialysis, and one ICU patient. Nine isolates were Enterococcus faecalis (seven positive for vanB and two negative for all three resistance genes) and three were Enterococcus faecium (all positive for vanB). Eight were high-level gentamicin resistant. PFGE suggested genetic clonality between the index isolate and five other isolates from renal patients. No specific clinical practice was associated with VRE colonisation. Attempts to clear rectal carriage with oral ampicillin/amoxycillin or bacitracin were of limited success. Although antibiotic prescribing in the Renal Unit was generally consistent with defined protocols, use of vancomycin and third-generation cephalosporins has been further restricted. CONCLUSIONS: Renal inpatients in our institution appear most at risk of VRE colonisation (4.6% overall) and therefore of VRE infection. Routine screening, especially of potentially high-risk patients, should be considered in major Australian hospitals.

Adult↗

Tuberculin-purified protein derivative-, MPT-64-, and ESAT-6-stimulated gamma interferon responses in medical students before and after Mycobacterium bovis BCG vaccination and in patients with tuberculosis.

QuantiFERON-TB (QIFN) (CSL Limited) is a whole-blood assay for the recognition of infection with Mycobacterium tuberculosis. QIFN measures gamma interferon (IFN-gamma) production when purified protein derivatives (PPDs) of mycobacteria are incubated with venous blood samples. The specificity of QIFN in medical students before and after BCG immunization was assessed, and sensitivity in patients with tuberculosis was assessed. Antigens were PPD derived from M. tuberculosis and two M. tuberculosis-specific proteins, ESAT-6 and MPT-64. Of 60 medical students, all of whom had 0-mm tuberculin skin tests (TSTs) at study entry, 58 (97%) were initially classified as negative for M. tuberculosis infection by PPD QIFN. Five months after BCG immunization, 7 of 54 students (13%) had a TST result of >/=10 mm and 11 of 54 students (20%) tested positive by PPD QIFN. ESAT-6- and MPT-64-stimulated IFN-gamma responses in the medical students were negative prior to and after BCG immunization. For patients with active tuberculosis, 12 of 19 (63%) were positive by PPD QIFN, 11 of 19 (58%) were positive by ESAT-6 QIFN, and 0 of 12 were positive by MPT-64 QIFN. In conclusion, PPD QIFN was negative in 97% of a low-risk population who had not received BCG and who had negative TSTs. The specificities of both the TST and PPD QIFN were reduced following BCG immunization. PPD QIFN and ESAT-6 QIFN were of similar and moderate sensitivity in patients with active tuberculosis, but ESAT-6 QIFN is likely to be more specific because it is not influenced by past BCG exposure.

Antibodies, Bacterial↗

Deletion of Escherichia coli groEL is complemented by a Rhizobium leguminosarum groEL homologue at 37 degrees C but not at 43 degrees C.

Bacterial Cpn60 proteins (homologues to the Escherichia coli GroEL protein) are often examined for function by testing their ability to complement a temperature sensitive mutation in the E. coli groEL gene. Such tests suffer from two drawbacks: the Cpn600 protein may come from a strain with a lower optimum growth temperature than E. coli, and the requirements for successful complementation in E. coli are likely to be more stringent at 43 degrees C than at lower temperatures. Here we describe the construction of a strain of E. coli where the chromosomal gene for the essential molecular chaperone GroEL has been deleted, with GroEL being expressed from a tightly regulated plasmid borne copy of the gene. The deletion can be transduced into strains expressing heterologous Cpn60 proteins, to test for complementation at any temperature. We show that a Cpn60 protein from the bacterium Rhizobium leguminosarum can function to allow E. coli growth at 37 degrees C but not at 43 degrees C. By switching off the plasmid borne groEL gene, the effects of progressive depletion of GroEL protein from E. coli cells can also be monitored at any temperature.

Chaperonin 60↗

The postantibiotic effect of imipenem: relationship with drug concentration, duration of exposure, and MIC.

The postantibiotic effect (PAE) of imipenem against Escherichia coli was measured at a wide variety of drug concentrations and times of exposure. We observed that the area under the concentration-time curve of drug exposure (AUC), the product of time of exposure and concentration of drug, is a much better predictor of the duration of the PAE than either parameter alone. We also measured the PAE of imipenem against strains of gram-positive and gram-negative bacteria for which MICs varied widely. The E50, the AUC required to produce 50% of the maximum PAE, is correlated with the MIC and is independent of species. This may explain why the duration of the PAE differs for bacteria of the same species for which MICs are different.

Area Under Curve↗

Disk diffusion interpretive criteria for fusidic acid susceptibility testing of staphylococci by the National Committee for Clinical Laboratory Standards method.

Fusidic acid is used in many countries for the treatment of multiresistant staphylococcal infection, especially multiresistant Staphylococcus aureus infection (MRSA). We collected consecutive fusidic acid-resistant isolates of staphylococci from the routine laboratory over several years, and compared these strains with fusidic acid-susceptible staphylococci to establish interpretive criteria for disk diffusion testing by National Committee for Clinical Laboratory Standards (NCCLS) methods. The minimum inhibitory concentrations (MICs) and zone diameters for strains of S. aureus (n = 102), including MRSA, S. saprophyticus (n = 20) and other coagulase-negative staphylococci (n = 115) were determined by NCCLS agar dilution and disk diffusion tests using a 2.5-micrograms disk of fusidic acid. MICs were bimodally distributed. No isolates had MICs of 0.5 or 1 microgram/ml; thus, we chose these values to define strains of intermediate susceptibility. The error-rate-bounded method was used to determine interpretive zone diameters for disk testing. Interpretive zone diameter criteria were found to be: susceptible > or = 22 mm, intermediate 18-21 mm, and resistant < or = 17 mm. All S. saprophyticus were intrinsically resistant to fusidic acid (MIC > or = 2 micrograms/ml).

Anti-Bacterial Agents↗

Diagnosing non-gonococcal urethritis: the gram-stained urethral smear in perspective.

The value of microscopy of a Gram-stained urethral smear in the diagnosis of non-gonococcal urethritis and chlamydial urethral infection was assessed in 153 men attending a department of genitourinary medicine. A mean of more than 4 polymorphs in 5 fields (x1000) was considered to be abnormal. In the diagnosis of urethritis, the sensitivity (94%), specificity (91%), positive predictive value (93%) and negative predictive value (93%) were high. In the diagnosis of chlamydial infection, sensitivity (91%) and negative predictive value (96%) were high, but specificity (68%) and positive predictive value (46%) were low. In men with symptoms of genitourinary infection, the sensitivity (96.7%) and negative predictive value (97.4%) of the Gram-stained urethral smear in the diagnosis of chlamydial infection was comparable with modern chlamydial antigen detection tests.

Chlamydia Infections↗

Fecal colonization with vancomycin-resistant enterococci in Australia.

To assess the rate of fecal vancomycin-resistant enterococci (VRE) colon ization in Austalia, we examined specimens from 1,085 healthy volunteers. VRE was cultured from 2(0.2%) of 1,085 specimens; both were vanB Enter ococcus faecium, identical by pulsed-field gel electrophoresis, but with a pattern rare in Melbourne hospitals.

Adult↗