Quality assessment of Microbe Base antimicrobial susceptibility data.

AIMS: To assess the quality of centres contributing antimicrobial susceptibility data to a centralised database. METHODS: Twelve organisms were distributed to 31 regional microbiology laboratories contributing data to a centralised susceptibility database. Participants were asked to determine susceptibilities to certain antibiotics by their routine method and return the data to the Department of Microbiology, Royal Hallamshire Hospital, Sheffield, for analysis. RESULTS: Results for the overwhelming majority of organism/antibiotic combinations were in agreement with expected results. Reasons for discrepancies included the non-bimodal distribution of susceptibilities, the use of different content discs, and, more importantly, minimum inhibitory concentrations falling close to breakpoint values. CONCLUSIONS: It is inevitable that any large multicentre database will contain a degree of inaccurate data. This study has highlighted several areas where discrepant results have occurred and has enabled Glaxo Laboratories to approach individual laboratories to address this problem. This study emphasises the value and consistency of Microbe Base as the largest database, of its kind, nationally.

Multipoint identification of Enterobacteriaceae: report of the British Society for Microbial Technology collaborative study.

AIMS: To evaluate the accuracy and reproducibility of multipoint identification schemes in a multicentre trial. METHODS: Forty two strains of Enterobacteriaceae were distributed to 22 laboratories for identification by routine multipoint methods. Analysis of results enabled inter- and intralaboratory reproducibility of a variety of tests, and the ability of laboratories to identify individual organisms to be determined. RESULTS: Interlaboratory reproducibility of most of the biochemical tests was acceptable. The least reproducible tests, both within and between laboratories, were citrate utilisation, production of urease and beta galactosidase, detection of motility, and decarboxylation of lysine and ornithine. Inconsistent results for these tests were often associated with misidentified strains. Most laboratories performed identifications satisfactorily. Most isolates (72.1%) were identified correctly to species level; 9.6% were incorrectly identified, and 6.4% could not be identified at all. The most difficult organisms to identify were Citrobacter freundii, Enterobacter cloacae, Hafnia alvei and Aeromonas hydrophila. Strains of Enterobacter, Serratia sp, and Providencia sp were difficult to speciate. Several laboratories could not identify organisms exhibiting at least one atypical biochemical reaction. CONCLUSION: This study emphasises the need for quality control of media and reagents for multipoint identification of Gram negative enteric bacilli.

Comparison of techniques for antimicrobial susceptibility testing of mycobacteria.

AIMS: To evaluate adenosine triphosphate (ATP) bioluminescence as a rapid technique for antimicrobial susceptibility testing of Mycobacterium spp by comparing it with conventional and radiometric methods, and to assess its potential for use in clinical microbiology laboratories. METHODS: 115 clinical isolates from a wide range of mycobacterial species and four control organisms of known susceptibility were tested against six antimicrobial agents. Minimum inhibitory concentrations (MICs) were determined after 4-6 weeks' incubation on Middlebrook 7H10 agar. Susceptibility was also determined radiometrically using a Bactec 460, and by bioluminescent assay of ATP using a 1250 luminometer (LKB-Wallac). RESULTS: Susceptibility results after 7 days showed excellent correlation with conventionally determined MICs. 714 susceptibility tests were performed by both techniques, with seven major discrepancies between the two systems. For pyrazinamide, agreement was 100%, but five strains of M tuberculosis, including one control, and 11 mycobacteria other than M tuberculosis (MOTT) failed to grow on Middlebrook agar at pH 5.5. 606 tests were performed by radiometry, with four major discrepancies between this technique and ATP bioluminescence. No particular species of Mycobacterium gave aberrant results. Contamination was a problem; 12 of the 119 strains tested were contaminated at day 1 and had to be repeated before results were obtained. Contamination of individual tests increased significantly after 7 days of incubation. CONCLUSIONS: ATP bioluminescence can be used to monitor mycobacterial growth in fluid culture media; the technique has considerable potential for rapid susceptibility testing. Advantages include lower initial cost of analytical equipment, lower reagent cost per test, and the use of non-radioactive substrates.

True identity of control Staphylococcus aureus strains and their performance in the tube coagulase test.

One hundred laboratories were asked to submit their control Staphylococcus aureus strains to determine the true identity of strains presumed to be S. aureus NCTC 6571, and also to evaluate the performance of those strains being used as controls in the tube coagulase test (TCT). Of the 60 who replied, 55 laboratories sent at least one strain labelled as S. aureus NCTC 6571 (total of 64 strains). Of these, 84% were identified as S. aureus, and were indistinguishable from a fresh type strain by a combination of phenotypic methods including biotyping, antibiotic susceptibility testing and phage typing. Six-to-ten strains (9-16%), depending on the degree of stringency, were not identifiable as S. aureus NCTC 6571. The time since last retrieval from storage ranged from daily to > or = 3 years, but there was no correlation between this duration and the likelihood of differing from S. aureus NCTC 6571. Forty-seven laboratories submitted 51 strains used as controls in the TCT; these included 31 strains labelled as S. aureus NCTC 6571, eight wild strains, three other NCTC strains and nine strains of uncertain origin. Generally, the S. aureus NCTC 6571 strains produced weaker clots than the remainder. None of the S. aureus NCTC 6571 strains was found to be inoculum dependent but four of the other control strains were. The study demonstrates that some laboratories must improve procedures for ensuring that control S. aureus strains retain their true identity, particularly by avoiding repeated subcultures. Laboratories are divided in their use of strong or weak (S. aureus NCTC 6571) positive controls for the TCT. S. aureus NCTC 6571 is a more stringent control for the TCT than other control strains presently being used and is, therefore, to be preferred.

Antimicrobial susceptibility testing of mycoplasmas by ATP bioluminescence.

The susceptibility of 72 mycoplasmas to a range of antimicrobial agents was assessed in a 6-h ATP bioluminescence system. ATP was assayed with the Amerlite Analyser. Correlation with conventionally determined MICs was excellent for erythromycin and tetracycline even at 3 h. However, for ciprofloxacin, correlation was poor unless incubation was extended to 6 h.

A numerical taxonomic study of the "Streptococcus milleri" group based upon conventional phenotypic tests and pyrolysis mass spectrometry.

Clinical strains presumptively identified as Streptococcus milleri (60), and blind coded collection strains (21) were characterised in conventional tests and pyrolysis mass spectrometry. Comparison of the clusters found by these two approaches revealed five clearly distinct centres of variation. Three corresponded to the DNA homology groups suggested by Whiley and Hardie (1989) as representing the species S. anginosus, S. intermedius and S. constellatus; a fourth comprised three Lancefield group C beta-haemolytic strains; the fifth may represent a biotype of S. anginosus. The characteristics of the latter group are described.

Mycoplasmas of the human genital tract.

The role of genital mycoplasmas in human infection is reviewed. Their isolation, identification and antimicrobial susceptibility are also discussed.

Evaluation of a commercial automated system for the identification of gram-negative enteric bacilli.

A total of 908 distinct clinical isolates and 60 reference strains of aerobic gram-negative bacilli were identified by our own in-house biochemical identification system (RHH) and by a commercial automated system (Mastascan Colour). Overall, both systems performed well in the identification of routine isolates of aerobic gram-negative bacilli, with only six discrepancies between the two systems. These six organisms were species infrequently encountered in the clinical microbiology laboratory. Of the 60 reference strains, many of which were biochemically atypical, the RHH system was unable to identify one and mis-identified two others. The commercial system was unable to identify one strain and misidentified five others. Both systems were inexpensive in terms of consumable materials, and the commercial system was compatible with the routine work of the department.

In-vitro selection of bacteria resistant to the 4-quinolone agents.

The activity of five 4-quinolone agents: norfloxacin, ciprofloxacin, ofloxacin, enoxacin and pefloxacin was tested against 121 distinct clinical bacterial isolates. Ciprofloxacin was the most active agent against Gram-negative bacteria and streptococci and pefloxacin was the most active against staphylococci. By serial sub-culture in sub-inhibitory concentrations of antibiotic we were able to select resistance in almost all Gram-positive and Gram-negative species, with the exception of Escherichia coli. Resistance to norfloxacin was more readily produced than resistance to the other four agents. There was almost complete cross-resistance among the five agents tested. The proportional increases in MIC were higher in Gram-negative than in Gram-positive organisms.

A 10 year survey of the antimicrobial susceptibility of urinary tract isolates in the UK: the Microbe Base project.

Microbe Base is a national computerized database comprising in excess of 1.7 million patient records down-loaded from the laboratory computer systems of 61 participating UK laboratories over 10 years. This paper highlights the antimicrobial susceptibilities of organisms isolated from the urinary tract which comprise around 50% of all isolates in the database. These data may be used to determine trends in antimicrobial susceptibilities; to formulate local antibiotic policies; to compare local with national data and, overall, to assist clinicians in the rational choice of antibiotic therapy and to prevent misuse, or overuse, of antibiotics.