Reproducibility of Control Strains for Antibiotic Susceptibility Testing

Abstract

Inter-and intralaboratory reproducibility of susceptibility testing requires stable control strains. The Food and Drug Administration diffusion procedure recommends the Seattle strains of Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) for this purpose. It was of interest to determine the present reproducibility of control cultures maintained in various laboratories over several years. Fifteen cultures each of S. aureus and E. coli were obtained from laboratories in different parts of the country. Their performance was compared with strains directly derived from ATCC. Diffusion susceptibility tests using a modified overlay technique were made with four replicates. Seven of the eight statistically significant differences in responses of the staphylococci were to penicillin, methicillin, or cephalothin. One culture was a penicillinase producer with a zone 15 mm less than the standard strain. Eleven of the 15 cultures showed no significant deviations or differences greater than 2 mm from the results with the strain derived directly from ATCC. All except the penicillinase producer were of identical phage type. Among 150 organism-antibiotic combinations tested with E. coli, all but one reading were within 2 mm of the standard. Four of the six statistically significant differences were in a culture from one laboratory. The stability of the cultures appears to have been influenced by the method of storage. Cultures that were kept frozen during extended storage were remarkably stable. Significant differences were found in cultures from four of five laboratories that maintained cultures in refrigerators or at ambient temperature.Recommended quality control procedures for antimicrobial disk susceptibility tests require daily testing of Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) (3, 4). These strains, originally isolated and used for quality control purposes in the University of Washington Hospital Clinical Laboratories, have now been maintained in many different laboratories for up to 7 years. This has raised the possibility that contamination or genetic drift may have occurred in some clones similar to that which was shown with the Oxford Staphylococcus by Oeding and Ostervold (5). A study was therefore designed to determine whether any differences could be detected in control strains used routinely in clinical laboratories.Laboratories in different parts of the country were requested to send working cultures of their control strains with information on their original source, the date obtained, and maintenance procedures used. These cultures were then compared with strains derived directly from the American Type Culture Collection using a closely standardized agar overlay diffusion procedure.MATERIALS AND METHODS Cultures. Cultures of E. coli and S. aureus submitted by 15 laboratories were tested. All were said to be derived from E. coli (ATCC 25922) and S. aureus (ATCC 25923) or from the "Seattle" control strains, which were the original source of the ATCC strains. The stan...

Bactericidal and Cytotoxic Activities of Polyphenol Extracts fromSolanum tuberosumspp.tuberosumand spp.andigenaCultivars onEscherichia coliand Human Neuroblastoma SH-SY5Y CellsIn Vitro

“…Two E. coli strains were used for experiments. E. coli ATCC 25922 is a recommended reference strain for antibiotic susceptibility testing [ 29 ]. E. coli O157 is a pathogenic strain that causes food intoxication by verotoxin production [ 30 ].…”

Neutrophil Microbicides Induce a Pathogen Survival Response in Community-Associated Methicillin-Resistant Staphylococcus aureus

“…To generate an isogenic isdAB mutant strain in MW2 (⌬isdAB), ⌬isdAB::ermC was transduced from strain Newman into MW2 by the transducing-phage -85 using a previously published protocol (19). S. aureus strains NCTC 12981 (ATCC 25923, originally isolated and used for quality control purposes at the University of Washington Clinical Laboratories in 1969) (20) and 502A (21) and serotype M1 group A Streptococcus (22) were published previously. For daily experiments, S. aureus cultures were seeded from overnight cultures using a dilution of 1/200 for MW2, LAC, 502A, and NCTC 12981; a 1/150 dilution for MRSA252; and a 1/100 dilution of COL. S. aureus were cultured at 37°C with shaking (250 rpm) to mid-exponential phase (OD 600 ϭ 0.75) of growth, at which time they were washed with Dulbecco's PBS (DPBS; Sigma-Aldrich) or RPMI 1640 medium (Invitrogen Life Technologies) buffered with 10 mM HEPES (RPMI/H, pH 7.2) and resuspended in the same buffer (typically prechilled at 4°C) at a cell density of 10 8 /ml.…”

The medically important aerobic actinomycetes: epidemiology and microbiology.

“…Several lyophilized systems have also been marketed; they differ only minimally in tray design, antimicrobial agents tested, and the type of inoculum and inoculation methods used. These commercial systems appear to provide very accurate and reproducible results when used in the manner specified by the manufacturers (125). Recently, the CDC Actinomycete Laboratory received several isolates of N. asteroides complex that did not grow in Mueller-Hinton broth.…”