Eighty-three isolates of Staphylococcus aureus for which MICs of methicillin of 4-16 mg/L had previously been recorded were tested for the presence of the mecA gene with a DNA probe and a PCR assay. There was complete agreement between the results obtained by these methods; 39 isolates were mecA-positive and 44 were mecA-negative. Using the presence of mecA as the defining standard, several phenotypic methods for determining resistance to methicillin were evaluated and a high-inoculum, agar-incorporation breakpoint test was found to offer the best combination of high sensitivity and high specificity. However twenty-seven of the 44 mecA-negative strains were methicillin-resistant according to agar dilution MICs (MIC > 4 mg/L on at least one of the four media used) but none had MICs exceeding 32 mg/L. One of the mecA-positive strains had a methicillin MIC of only 8 mg/L and did not appear to be heteroresistant. The clinical significance of these two groups of 'atypical' isolates may need further investigation. This study highlights the problems of detecting reliably S. aureus with low level methicillin resistance by phenotype methods and the usefulness of direct detection of the mecA gene.

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In the last 5 years, clinical isolates of gram-positive bacteria with intrinsic or acquired resistance to glycopeptide antibiotics have been encountered increasingly. In many of these isolates, resistance arises from an alteration of the antibiotic target site, with the terminal D-alanyl-D-alanine moiety of peptidoglycan precursors being replaced by groups that do not bind glycopeptides. Although the criteria for defining resistance have been revised frequently, the reliable detection of low-level glycopeptide resistance remains problematic and is influenced by the method chosen. Glycopeptide-resistant enterococci have emerged as a particular problem in hospitals, where in addition to sporadic cases, clusters of infections with evidence of interpatient spread have occurred. Studies using molecular typing methods have implicated colonization of patients, staff carriage, and environmental contamination in the dissemination of these bacteria. Choice of antimicrobial therapy for infections caused by glycopeptide-resistant bacteria may be complicated by resistance to other antibiotics. Severe therapeutic difficulties are being encountered among patients infected with enterococci, with some infections being untreatable with currently available antibiotics.

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Objective-To assess the prevalence of antibiotic resistance and serotype distribution among pneumococci in England and Wales in 1990 and 1995.Design-Observational surveys in March 1990 and March 1995. During two weeks in each survey period all pneumococci isolated in public health laboratories in England and Wales were collected and assessed for sensitivity to antibiotics and the distribution of serogroups or serotypes.Setting-The network ofpublic health laboratories throughout England and Wales.

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Infective episodes in immunocompromised children with indwelling central venous catheters were studied prospectively for one year. Culture of catheter hubs and skin at catheter entry sites during the first six months suggested that hub contamination was important in the pathogenesis of catheter colonization. The incidence of catheter-related bacteraemia, and possible catheter-related bacteraemia, fell by 56.5% following alterations in the protocol for manipulative care of catheters, from 5.82 per 1000 catheter days in the first six months to 2.53 per 1000 catheter days in the subsequent six months. A firm diagnosis of catheter-related bacteraemia was made simply and economically by a pour-plate quantitative blood culture technique. Attempts at eradication of catheter-related bacteraemia without removal of the catheter were successful in all cases.