The National Committee for Clinical Laboratory Standards has recently changed the oxacillin breakpoint from ≥4 mg/liter to ≥0.5 mg/liter to detect methicillin-resistant coagulase-negative staphylococci (CoNS) because the previous breakpoint lacked sensitivity. To determine the correlation between the new oxacillin breakpoint and the presence of themecA gene, 493 CoNS of 11 species were tested. The presence of the mecA gene was determined by PCR, and oxacillin susceptibility was determined by the agar dilution method with Mueller-Hinton agar containing 2% NaCl and oxacillin (0.125 to 4.0 mg/liter). The new breakpoint correctly classified all CoNS strains with mecA as methicillin resistant and strains ofStaphylococcus epidermidis, S. haemolyticus, and S. hominiswithout mecA as methicillin susceptible. The breakpoint of ≥0.5 mg/liter was not specific for S. cohnii, S. lugdunensis, S. saprophyticus, S. warneri, and S. xylosus, in that it categorized 70 of 74 strains of these species withoutmecA (94.6%) as methicillin resistant. The results of this study indicate that the new oxacillin breakpoint accurately identifies strains of CoNS with mecAbut is not specific for strains of certain species of CoNS withoutmecA.

A severe outbreak of hemorrhagic colitis occurred in London, Ontario, during the month of September 1985. A total of 55 residents and 18 employees of a nursing home developed diarrhea, and 17 residents (age range, 78 to 99 years) died. Specimens from 38 patients, 37 employees and contacts, and 10 autopsies were investigated for all enteric pathogens. Specimens were also planted on MacConkey-sorbitol agar. Fecal extracts were tested on Vero cells for cytotoxin (FVT). Escherichia coli isolates were serotyped and tested for verotoxin and beta-glucuronidase production. Of the 38 symptomatic patients, 26 were positive for FVT, verotoxin-producing E. coli (VTEC), or both. Of the 105 specimens that were examined from these 38 patients, FVT and VTEC were both positive in 30 specimens, FVT only was positive in 13 specimens, and VTEC only was positive in 4 specimens. None of the 27 specimens from 10 autopsies was positive for FVT or VTEC. No other enteric pathogen was found in any of the cases. All asymptomatic individuals were negative for both FVT and VTEC. Of 19 VTEC strains that were isolated, 18 belonged to serotype 0157:H7. These 18 strains and 2 more strains that were obtained from sporadic cases that had occurred within the 2 previous months were found to give similar biochemical reactions in a 36-test identification system. All isolates of serotype 0157:H7 were beta-glucuronidase negative and susceptible to the antimicrobial agents that are used to treat E. coli infections. Testing for FVT and VTEC was found to be the most sensitive and specific technique for the laboratory diagnosis of this disease. Negative sorbitol, positive raffinose, and negative beta-glucuronidase tests appeared to be consistent markers for aiding in the detection of E. coli 0157:H7.

The methicillin resistance of 363 coagulase-negative staphylococci isolated from blood cultures was determined by a slide latex agglutination (LA) test for penicillin-binding protein 2a (PBP 2a), the presence of the mecA gene by PCR, disk diffusion, and Vitek. LA was performed on primary cultures, and PBP 2a expression was induced by placing an oxacillin disk in the primary inoculum. Compared to the PCR results, LA was the most sensitive and specific in the detection of methicillin resistance. Without induction, LA failed to detect 50% of mecA-positive strains grown on two different media.During the 1980s, the National Nosocomial Infection Surveillance surveys demonstrated the increasing significance of gram-positive organisms in hospital-acquired infections (3). Recent reports have confirmed that coagulase-negative staphylococci (CoNS) remain an important cause of bacteremia in hospitalized patients (1, 12). In North America over the last three decades, this increase in infections by staphylococci has been paralleled by an increase in resistance to methicillin among these organisms. In 1975, only 2% of Staphylococcus aureus strains were resistant to methicillin, while in 1991, 29% were resistant (4). Methicillin resistance is even more prevalent among CoNS, particularly with hospital-acquired infections (12).The increase in CoNS as a cause of bacteremia and the prevalence of methicillin resistance in these organisms have led to increased use of vancomycin as empirical therapy. Concern about the spread of vancomycin-resistant enterococci has prompted the Hospital Infection Control Advisory Committee to recommend that vancomycin be used judiciously (7). In addition, beta-lactam agents are considered a better choice for their therapeutic profile than vancomycin against oxacillinsusceptible staphylococci (12). Early identification of methicillin resistance in staphylococci, especially from blood isolates, could, therefore, curtail unnecessary use of vancomycin and allow earlier optimal therapy of infections in some cases.The MRSA Screen latex agglutination (LA) test (Denka Seiken, Niigata, Japan) is a simple slide agglutination test designed to detect the presence of penicillin-binding protein 2a (PBP 2a). It has been shown to be reliable for the detection of oxacillin resistance in CoNS (10). In this study, we evaluated the usefulness of this test for detecting the antibiotic susceptibilities of CoNS isolated from blood cultures by performing the test on the primary isolates with a rapid turnaround time.Blood cultures submitted to the microbiology laboratory were processed with the BacT/Alert system (Organon Teknika Corporation, Durham, N.C.). If gram-positive cocci in clusters were seen on a Gram stain, the blood culture was subcultured onto Columbia agar (BA; Oxoid, Napean, Ontario, Canada) with 5% sheep blood. A 1-g oxacillin disk was placed in the primary inoculum. After overnight incubation, if the isolate was identified as Staphylococcus, growth around the disks was used to perform the LA test.The LA test wa...

A rapid slide latex agglutination (LA) test, MRSA-Screen (Denka Seiken Co., Niigata, Japan), which detects PBP 2a, was tested for its ability to differentiate between mecA-positive and -negative coagulase-negative staphylococci. A total of 463 isolates from 13 species were included in the study. The mecA gene was detected by PCR, and the oxacillin MIC was determined by the agar dilution method according to the guidelines of the National Committee for Clinical Laboratory Standards (NCCLS). The LA test was performed with oxacillininduced isolates. The true-positive and true-negative results were defined on the basis of the presence or the absence of the mecA gene. By PCR, 251 isolates were mecA positive and 212 were mecA negative. The sensitivities, specificities, and positive and negative predictive values for the LA test compared to the NCCLS breakpoint for oxacillin resistance (>0.5 mg/liter) were as follows: for the LA test, 100, 99.5, 99.6, and 100%, respectively; for the NCCLS breakpoint, 100, 60.8, 75.1, and 100%, respectively. One hundred twenty-five mecA-positive isolates were also tested by the LA test without induction of PBP 2a; only 72 (57.6%) gave a positive result and required 3 to 15 min for reaction. With induction, all 251 isolates were positive within 3 min. The LA test was reliable in classifying mecA-negative isolates, but it classified isolates for which the oxacillin MIC was >0.5 mg/liter as oxacillin susceptible. For the reliable detection of oxacillin resistance by the MRSA-Screen in coagulase-negative staphylococci, induction of the mecA gene appears to be necessary.

A rapid slide latex agglutination (LA) test, MRSA-Screen (Denka Seiken Co., Niigata, Japan), which detects PBP 2a, was tested for its ability to differentiate between mecA-positive and-negative coagulase-negative staphylococci. A total of 463 isolates from 13 species were included in the study. The mecA gene was detected by PCR, and the oxacillin MIC was determined by the agar dilution method according to the guidelines of the National Committee for Clinical Laboratory Standards (NCCLS). The LA test was performed with oxacillininduced isolates. The true-positive and true-negative results were defined on the basis of the presence or the absence of the mecA gene. By PCR, 251 isolates were mecA positive and 212 were mecA negative. The sensitivities, specificities, and positive and negative predictive values for the LA test compared to the NCCLS breakpoint for oxacillin resistance (>0.5 mg/liter) were as follows: for the LA test, 100, 99.5, 99.6, and 100%, respectively; for the NCCLS breakpoint, 100, 60.8, 75.1, and 100%, respectively. One hundred twenty-five mecA-positive isolates were also tested by the LA test without induction of PBP 2a; only 72 (57.6%) gave a positive result and required 3 to 15 min for reaction. With induction, all 251 isolates were positive within 3 min. The LA test was reliable in classifying mecA-negative isolates, but it classified isolates for which the oxacillin MIC was >0.5 mg/liter as oxacillin susceptible. For the reliable detection of oxacillin resistance by the MRSA-Screen in coagulase-negative staphylococci, induction of the mecA gene appears to be necessary.