Public Health Efforts Can Impact Adoption of Current Susceptibility Breakpoints, but Closer Attention from Regulatory Bodies Is Needed.

Microbiological testing, including interpretation of antimicrobial susceptibility testing results using current breakpoints, is crucial for clinical care and infection control. Continued use of obsolete Enterobacteriaceae carbapenem breakpoints is common in clinical laboratories. The purposes of this study were (i) to determine why laboratories failed to update breakpoints and (ii) to provide support for breakpoint updates. The Los Angeles County Department of Public Health conducted a 1-year outreach program for 41 hospitals in Los Angeles County that had reported, in a prior survey of California laboratories, using obsolete Enterobacteriaceae carbapenem breakpoints. In-person interviews with hospital stakeholders and customized expert guidance and resources were provided to aid laboratories in updating breakpoints, including support from technical representatives from antimicrobial susceptibility testing device manufacturers. Forty-one hospitals were targeted, 7 of which had updated breakpoints since the prior survey. Of the 34 remaining hospitals, 27 (79%) assumed that their instruments applied current breakpoints, 17 (50%) were uncertain how to change breakpoints, and 10 (29%) lacked resources to perform a validation study for off-label use of the breakpoints on their systems. Only 7 hospitals (21%) were familiar with the FDA/CDC Antibiotic Resistance Isolate Bank. All hospitals launched a breakpoint update process; 16 (47%) successfully updated breakpoints, 12 (35%) received isolates from the CDC in order to validate breakpoints on their systems, and 6 (18%) were planning to update within 1 year. The public health intervention was moderately successful in identifying and overcoming barriers to updating Enterobacteriaceae carbapenem breakpoints in Los Angeles hospitals. However, the majority of targeted hospitals continued to use obsolete breakpoints despite 1 year of effort. These findings have important implications for the quality of patient care and patient safety. Other public health jurisdictions may want to utilize similar resources to bridge the patient safety gap, while manufacturers, the FDA, and others determine how best to address this growing public health issue.

Evaluation of Oxacillin and Cefoxitin Disk Diffusion and MIC Breakpoints Established by the Clinical and Laboratory Standards Institute for Detection of mecA-Mediated Oxacillin Resistance in Staphylococcus schleiferi.

Staphylococcus schleiferi is a beta-hemolytic, coagulase-variable colonizer of small animals that can cause opportunistic infections in humans. In veterinary isolates, the rate of mecA-mediated oxacillin resistance is significant, with reported resistance rates of >39%. The goal of this study was to evaluate oxacillin and cefoxitin disk diffusion (DD) and MIC breakpoints for detection of mecA-mediated oxacillin resistance in 52 human and 38 veterinary isolates of S. schleiferi Isolates were tested on multiple brands of commercial media and according to Clinical and Laboratory Standards Institute (CLSI) methods. Zone diameters and MIC values were interpreted using CLSI breakpoints (CLSI, Performance Standards for Antimicrobial Susceptibility Testing. M100-S27, 2017) for Staphylococcus aureus/Staphylococcus lugdunensis, coagulase-negative staphylococci (CoNS), and Staphylococcus pseudintermedius Results were compared to those of mecA PCR. Twenty-nine of 90 (32%) isolates were mecA positive. Oxacillin inhibition zone sizes and MICs interpreted by S. pseudintermedius breakpoints reliably differentiated mecA-positive and mecA-negative isolates, with a categorical agreement (CA) of 100% and no very major errors (VMEs) or major errors (MEs) for all media. For cefoxitin DD results interpreted using S. aureus/S. lugdunensis and CoNS breakpoints, CA values were 85% and 75%, respectively, and there were 72% and 64% VMEs, respectively, and 0 MEs. For cefoxitin MICs interpreted using S. aureus/S. lugdunensis breakpoints, CA was 81%, and there were 60% VMEs and no MEs. Our data demonstrate that oxacillin DD or MIC testing methods using the current S. pseudintermedius breakpoints reliably identify mecA-mediated oxacillin resistance in S. schleiferi, while cefoxitin DD and MIC testing methods perform poorly.

Emerging Resistance, New Antimicrobial Agents     but No Tests! The Challenge of Antimicrobial Susceptibility Testing in the Current US Regulatory Landscape.

Accurate and timely performance of antimicrobial susceptibility testing (AST) by the clinical laboratory is paramount to combating antimicrobial resistance. The ability of laboratories in the United States to effectively perform ASTs is challenged by several factors. Some, such as new resistance mechanisms and the associated evolution of testing recommendations and breakpoints, are inevitable. Others are entirely man-made. These include unnecessarily strict US Food and Drug Administration (FDA) limitations on how commercial AST systems can be used for diagnostic testing, the absence of up-to-date performance data on these systems, and the lack of commercially available FDA-cleared tests for newer antimicrobial agents or for older agents with updated breakpoints. This viewpoint will highlight contemporary AST challenges faced by the clinical laboratory, and propose some solutions.

Multisite Evaluation of Cepheid Xpert Carba-R Assay for Detection of Carbapenemase-Producing Organisms in Rectal Swabs.

Rapid identification of patients who are colonized with carbapenemase-producing organisms (CPO) is included in multiple national guidelines for containment of these organisms. In a multisite study, we evaluated the performance of the Cepheid Xpert Carba-R assay, a qualitative diagnostic test that was designed for the rapid detection and differentiation of the blaKPC, blaNDM, blaVIM, blaOXA-48, and blaIMP-1 genes from rectal swab specimens. A double rectal swab set was collected from 383 patients admitted at four institutions (2 in the United States, 1 in the United Kingdom, 1 in Spain). One swab was used for reference culture (MacConkey broth containing 1 mg/liter of meropenem and subcultured to a MacConkey agar plate with a 10-µg meropenem disk) and for sequencing of DNA obtained from carbapenem-nonsusceptible isolates for carbapenemase identification. The other swab was used for the Xpert Carba-R assay. In addition to the clinical rectal swabs, 250 contrived specimens (108 well-characterized CPO and 142 negative controls spiked onto negative rectal swabs) were tested. Overall, 149/633 (23.5%) samples were positive by the Xpert Carba-R assay. In 6 samples, multiple targets were detected (4 VIM/OXA-48, 1 IMP-1/NDM, and 1 NDM/KPC). The Xpert Carba-R assay detected 155 targets (26 IMP-1, 30 VIM, 27 NDM, 33 KPC, 39 OXA-48) within a time range of 32 to 48 min. The sensitivity, specificity, and positive and negative predictive values of the Xpert Carba-R assay compared to those of the reference culture and sequencing results were 96.6% (95% confidence interval [CI], 92.2% to 98.9%), 98.6% (95% CI, 97.1% to 99.4%), 95.3%, and 99.0%, respectively. The Cepheid Xpert Carba-R assay is an accurate and rapid test to identify rectal colonization with CPO, which can guide infection control programs to limit the spread of these organisms.

Evaluation of Oxacillin and Cefoxitin Disk and MIC Breakpoints for Prediction of Methicillin Resistance in Human and Veterinary Isolates of Staphylococcus intermedius Group.

Staphylococcus pseudintermedius is a coagulase-positive species that colonizes the nares and anal mucosa of healthy dogs and cats. Human infections with S. pseudintermedius range in severity from bite wounds and rhinosinusitis to endocarditis; historically, these infections were thought to be uncommon, but new laboratory methods suggest that their true incidence is underreported. Oxacillin and cefoxitin disk and MIC tests were evaluated for the detection of mecA- or mecC-mediated methicillin resistance in 115 human and animal isolates of the Staphylococcus intermedius group (SIG), including 111 Staphylococcus pseudintermediusand 4 Staphylococcus delphini isolates, 37 of which were mecA positive. The disk and MIC breakpoints evaluated included the Clinical and Laboratory Standards Institute (CLSI) M100-S25 Staphylococcus aureus/Staphylococcus lugdunensis oxacillin MIC breakpoints and cefoxitin disk and MIC breakpoints, the CLSI M100-S25 coagulase-negative Staphylococcus (CoNS) oxacillin MIC breakpoint and cefoxitin disk breakpoint, the CLSI VET01-S2 S. pseudintermedius oxacillin MIC and disk breakpoints, and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) S. pseudintermedius cefoxitin disk breakpoint. The oxacillin results interpreted by the VET01-S2 (disk and MIC) and M100-S25 CoNS (MIC) breakpoints agreed with the results of mecA/mecC PCR for all isolates, with the exception of one false-resistant result (1.3% of mecA/mecC PCR-negative isolates). In contrast, cefoxitin tests performed poorly, ranging from 3 to 89% false susceptibility (very major errors) and 0 to 48% false resistance (major errors). BD Phoenix, bioMérieux Vitek 2, and Beckman Coulter MicroScan commercial automated susceptibility test panel oxacillin MIC results were also evaluated and demonstrated >95% categorical agreement with mecA/mecC PCR results if interpreted by using the M100-S25 CoNS breakpoint. The Alere penicillin-binding protein 2a test accurately detected all mecA-positive isolates, although for four isolates, cefoxitin induction was required prior to testing. These data demonstrate that the cefoxitin surrogate test does not reliably detect the presence of mecA in S. pseudintermedius isolates and that laboratories should perform oxacillin disk or MIC tests of these isolates when they are encountered.

Phenotypic and molecular characteristics of carbapenem-resistant Enterobacteriaceae in a health care system in Los Angeles, California, from 2011 to 2013.

Carbapenem-resistant Enterobacteriaceae (CRE) are a concern for health care in the United States but remain relatively uncommon in California. We describe the phenotype, clonality, and carbapenemase-encoding genes present in CRE isolated from patients at a Californian tertiary health care system. CRE for this study were identified by evaluating the antibiograms of Enterobacteriaceae isolated in the UCLA Health System from 2011 to 2013 for isolates that were not susceptible to meropenem and/or imipenem. The identification of these isolates was subsequently confirmed by matrix-associated laser desorption ionization-time of flight, and broth microdilution tests were repeated to confirm the CRE phenotype. Real-time PCR for bla(KPC), bla(SME), bla(IMP), bla(NDM-1), bla(VIM), and bla(OXA-48) was performed. Clonality was assessed by repetitive sequence-based PCR (repPCR) and multilocus sequence typing (MLST). Of 15,839 nonduplicate clinical Enterobacteriaceae isolates, 115 (0.73%) met the study definition for CRE. This number increased from 0.5% (44/8165) in the first half of the study to 0.9% (71/7674) in the second (P = 0.004). The most common CRE species were Klebsiella pneumoniae, Enterobacter aerogenes, and Escherichia coli. A carbapenemase-encoding gene was found in 81.7% (94/115) of CRE and included bla(KPC) (78.3%), bla(NDM-1) (0.9%), and bla(SME) (2.6%). The majority of bla(KPC) genes were in K. pneumoniae isolates, which fell into 14 clonal groups on typing. bla(KPC) was identified in more than one species of CRE cultured from the same patient in four cases. Three bla(SME)-carrying Serratia marcescens isolates and one bla(NDM-1) carrying Providencia rettgeri isolate were detected. CRE are increasing in California, and carbapenemases, particularly KPC, are a common mechanism for carbapenem resistance in this region.

Aminoglycoside resistance: a worldwide perspective.

Two concerns, neither of which is particularly new, underlie the current reluctance to use aminoglycosides more broadly. First, an undeniable fact is that these compounds can be toxic, particularly in patients with impaired renal function or those receiving other nephrotoxic medications. Second, a more emotional concern is that widespread use of aminoglycosides, particularly the newer compounds that are more resistant to enzymatic inactivation, may engender widespread resistance. In fact, several sources lead one to doubt whether widespread use of potent and highly effective agents like amikacin will by itself increase a clinical reservoir of more resistant microbes. First, the surveillance studies undertaken in many hospitals show some modest reduction in overall aminoglycoside resistance even when a drug like amikacin is used to supplant antecedent compounds of the same class. Second, in institutions where no official surveillance programs have been undertaken but where ongoing surveillance has been maintained, susceptibility to amikacin has remained constant when recent blood isolates are compared with blood isolates from more than 10 years ago. Third, in controlled clinical trials, particularly in immunocompromised patients, the overall emergence of resistance has been remarkably low and contrasts rather strikingly with what has been observed in some monotherapeutic studies of beta-lactam agents. The presence of aminoglycoside-resistant strains cannot be denied, but the circumstances leading to the emergence of such resistance must be carefully assessed, particularly outside of the setting in which these drugs are used as first-line therapy for critically ill patients. For instance, there is substantial evidence to suggest that the topical use of aminoglycosides or the use of these agents when there may be environmental contamination could lead to the emergence of resistance. Before one incriminates the use of any one drug as predisposing to the emergence of resistance, one needs to have more information about the total exposure of a given bacterial population to aminoglycoside therapy. The emergence of resistance to aminoglycosides has been associated with exposure to the more commonly used agents such as gentamicin or tobramycin. With some of the newer beta-lactam agents, the rate of emergence of resistance, unlike that of the aminoglycosides, has appeared to be remarkably high. If the concern about emergence of resistance is genuine, and to maintain consistency of approaches, the infectious disease community should focus more attention on limiting or restricting the use of the more widely used beta-lactam compounds.(ABSTRACT TRUNCATED AT 400 WORDS)

Laboratory-based surveillance for vancomycin-resistant enterococci: utility of screening stool specimens submitted for Clostridium difficile toxin assay.

OBJECTIVE: To study vancomycin-resistant enterococci (VRE) gastrointestinal colonization prevalence in high-risk hospitalized patients and to assess the cost and utility of this laboratory-based surveillance. SETTING: Large university teaching hospital. DESIGN: Quarterly prevalence culture survey of 50 stool specimens submitted for Clostridium difficile toxin A assay from October 1996 through June 1999 (n=526). Screening culture survey of all C difficile-positive stool specimens from July 1998 through June 1999 (n=140). PATIENTS: Specimens for analysis were collected from patients who were admitted to the hospital and who had C difficile toxin A testing ordered. Patient samples were excluded from analysis if they were obtained from patients not hospitalized at UCLA Medical Center, if the C difficile toxin assay result was indeterminate, or if the patient was known to have previous VRE colonization or infection. RESULTS: During quarterly surveillance, VRE was detected in 19.8%, C difficile toxin A in 9.5%, and both VRE and C difficile toxin A in 3.2% of stool specimens submitted for C difficile toxin assay. Patients whose stool specimens were positive for C difficile toxin A were significantly more likely than those whose specimens were negative to have VRE detected (odds ratio, 2.3; 95% confidence interval, 1.2-4.5). Based on these findings, in July 1998, we began routine screening of all C difficile-positive stool specimens for VRE. From July 1998 through June 1999, 58 (41.4%) of 140 patients with C difficile-positive specimens had VRE newly detected in the stool. The combined cost of the two laboratory-based surveillance strategies was approximately $62 per VRE-positive patient identified and $5,800 per year. CONCLUSION: Quarterly surveillance of stool submitted for C difficile assay combined with screening all C difficile-positive stools is a cost-effective and efficient strategy for detecting VRE stool colonization among high-risk hospitalized patients. Such a laboratory-based surveillance should be included as part of a comprehensive program to limit nosocomial VRE transmission.

DNA probe reactivity of Mycobacterium avium complex isolates from patients without AIDS.

Mycobacterium avium complex isolates from 27 patients without AIDS and from 76 patients with AIDS were analyzed with the Gen-Probe Rapid Diagnostic System for Mycobacterium avium complex, and a retrospective chart review was performed to determine clinical significance of the isolates. While 87% of isolates from AIDS patients reacted only with the M. avium probe, only 37% from non-AIDS patients were M. avium probe positive (p less than 0.001). This pattern among non-AIDS patients was also observed among the 13 patients from whom isolates were considered to be clinically significant. Reactivity to both probes occurred with three isolates, two from non-AIDS patients that were not clinically significant and one from an AIDS patient. Results of further testing suggested that these represented dual infection with two coexisting strains. Awareness of the differences in DNA probe reactivity between isolates from AIDS and non-AIDS patients may influence testing strategies in the clinical laboratory.

In vitro activity of CI-934 compared with ciprofloxacin, enoxacin, norfloxacin, and vancomycin.

The in vitro activity of the quinolone CI-934 was compared with ciprofloxacin, norfloxacin, enoxacin, and vancomycin against 607 Gram-positive and -negative isolates. CI-934 inhibited 90% of the Enterobacteriaceae, Aeromonas hydrophila, and Acinetobacter spp. at 1 microgram/ml. Decreased activity was observed against Pseudomonas aeruginosa. Pseudomonas maltophilia, and other Pseudomonas spp. with CI-934 MIC90 greater than or equal to 8 micrograms/ml. CI-934 activity against Gram-positive organisms exceeded vancomycin and the other quinolones. MIC90 for Streptococcus faecalis, Listeria monocytogenes, and Corynebacterium spp. were less than or equal to 2 micrograms/ml. CI-934 was equally effective against oxacillin-susceptible and -resistant staphylococci with MIC90 of 0.5 micrograms/ml.

Validation of NCCLS macrolide (azithromycin, clarithromycin, and erythromycin) interpretive criteria for Haemophilus influenzae tested with the Haemophilus test medium. National Committee for Clinical Laboratory Standards.

Some recently marketed macrolide antimicrobial agents possess physiochemical, antimicrobial, and pharmacokinetic advantages that enable their wider clinical use against Haemophilus influenzae infections. A five-laboratory study assessed the validity of existing or proposed azithromycin, clarithromycin, and erythromycin interpretive criteria for tests with H. influenzae isolates. National Committee for Clinical Laboratory Standards (NCCLS) methods, criteria, and quality-control guidelines were used. A total of 350 H. influenzae strains were processed, including fresh clinical isolates (250 strains) and replicate tests of 100 stock cultures sampling strains isolated from 1984 to 91. Azithromycin interpretive criteria (susceptible at < or = 4 micrograms/ml, > or = 12 mm) produced a 99.8% absolute agreement between the minimum inhibitory concentrations and disk diffusion results (0.2% false-susceptible error). Clarithromycin breakpoint criteria (susceptible at < or = 8 micrograms/ml, > or = 13 mm; and resistant at > or = 32 micrograms/ml, < or = 10 mm) produced high minor interpretive error, but < or = 1% combined false-susceptible and false-resistant discrepancies. Erythromycin interpretive guidelines were initially proposed for susceptible at < or = 0.5 microgram/ml, > or = 26 mm. This categorizes nearly all H. influenzae strains as resistant to this older macrolide. The NCCLS should consider the proposed erythromycin criteria for publication in appropriate tables, and a class drug should also be selected (azithromycin) that would best predict macrolide-class susceptibility for those agents indicated by the US Food and Drug Administration for H. influenzae infection chemotherapy (azithromycin and clarithromycin). No serious interpretive problems were observed with the current NCCLS criteria using Haemophilus test medium.

Clinical applicability of antifungal susceptibility testing on non-Candida albicans species in hospitalized patients.

We assessed the distribution, antifungal susceptibility, and treatment associated with 161 non-Candida albicans isolates recovered from hospitalized patients over a 6-month period. The three most prevalent species were C. glabrata (100), C. tropicalis (28), and C. krusei (15). Resistance of C. glabrata to fluconazole and itraconazole were 6% and 17% respectively; 80% of the fluconazole-resistant isolates were cross-resistant to itraconazole. Prior azole exposure significantly reduced azole susceptibility in C. glabrata and also affected its subsequent selection among colonized patients. Only 21% of the patients had clinical infections. Patients with fungemia were more likely to be treated with amphotericin versus an azole. Overall treatment success was higher in patients treated with amphotericin versus an azole (56% vs 31%). Routine susceptibility testing on all Candida species does not appear necessary except where therapy with an azole is being considered to detect resistant isolates or for epidemiologic surveillance purposes. Further studies are needed to delineate the relationship between azole MICs and treatment outcomes of invasive candidiasis due to non-C. albicans species.

Serum bactericidal activity from intravenous ciprofloxacin and azlocillin given alone and in combination to healthy subjects.

Ciprofloxacin plus azlocillin have been shown to exhibit in vitro synergy versus a variety of organisms, including Pseudomonas aeruginosa. This study examined this interaction in vivo, testing serum bactericidal activity (SBA) in six healthy male subjects after intravenous administration of ciprofloxacin 4 mg/kg (C), azlocillin 60 mg/kg (A), and the two simultaneously (C/A). Eight different organisms were tested: four isolates of P. aeruginosa with varying susceptibilities to C and A, and one isolate each of Escherichia coli (EC), Staphylococcus aureus (SA) Serratia marcescens (SM), and Klebsiella pneumoniae (KP), all of which were susceptible to both drugs. Blood samples were collected at the end of 30-min infusions and at 4 and 8 hr. Reciprocal titers were plotted versus time and area under the bactericidal titer curve (AUBC) calculated to assess antibacterial interactions. Results indicated that P. aeruginosa-1 (PA-1), EC, and KP were synergistically killed by C/A. AUBC for PA-1 were C = 36, A = 11, C/A = 144, p less than 0.05. AUBC for EC were C = 1059, A = 180, C/A = 1504, p = 0.05. AUBC for KP were C = 327, A = 97, C/A = 584, p = 005. Additive effects were demonstrated versus all of the other organisms except Serratia marcescens, where an indifferent effect was observed. Ciprofloxacin plus azlocillin may be a useful combination of the treatment of selected Gram-negative bacillary infections.

Comparative activity of twelve beta-lactam drugs tested against penicillin-resistant Streptococcus pneumoniae from five medical centers: effects of serum protein and capsular material on potency and spectrum as measured by reference tests.

A total of 152 strains of Streptococcus pneumoniae from diverse geographic areas in the United States and with different levels of penicillin resistance were tested against five broad-spectrum cephalosporins, ampicillin, piperacillin, ticarcillin, and three beta-lactamase inhibitor combinations. Also, the effect of human serum proteins on the activity of selected "third-generation" cephalosporins was examined. The overall rank order of activity among the cephalosporins against penicillin-susceptible strains was: ceftriaxone (MIC90, 0.03 microgram/mL) > cefotaxime > ceftizoxime = cefuroxime > ceftazidime (MIC90, 0.5 microgram/mL). Only cefotaxime and ceftriaxone exhibited significant activity against penicillin-intermediate or -resistant isolates. Ampicillin, piperacillin, and penicillin were generally eight- to 16-fold more potent than ticarcillin and no increase in the effectiveness of these agents was observed with the addition of the beta-lactamase inhibitors (clavulanate, sulbactam, tazobactam). Ceftriaxone potency was significantly decreased (> or = four-fold) by the modest addition of 25% pooled human serum proteins and this change modified the rank order of potency against nonpenicillin-susceptible pneumococci to favor cefotaxime (41% resistant versus 71% for ceftriaxone; MICs at > or = 2 micrograms/mL). Induced high-level capsular production had no measurable effect on the MIC results of tested agents. These results confirm the continued activity advantages of cefotaxime and ceftriaxone against various populations of pneumococci compared to other alternative beta-lactams. The predictive value, however, of the utilized breakpoint concentrations of the cephalosporins, remains in question for pneumococcal infections other than those in the central nervous system and at unaltered, "usual" dosing.

Parenteral infusions as culture media from a viewpoint of nosocomial bacteremia.

OBJECTIVE: To assess the growth patterns of selected organisms in common parenteral solutions, in order to ascertain implications for nosocomial bacteremia. DESIGN: A microbial suspension of approximately 300 CFU/mL was sequentially inoculated into common parenteral infusions from three different manufacturers and incubated at room temperature. Initially, 11 bacterial isolates and one Candida species from clinical specimens were studied. Eight gram-negative rods (GNR) were tested at varying pH's. Species variability was examined by testing an additional 39 isolates. RESULTS: The eight GNR grew in Ringer's lactate (RL) from two manufacturers and only two grew in dextrose 5% in water (D5/W) (Klebsiella pneumoniae and Serratia marcescens). No organism grew in saline or dextrose 5% in saline. The gram-positive cocci and Candida did not grow in any solution. No significant changes in growth were found after modifying the pH of solutions. Significant inter- and intra-species growth variability was noted. CONCLUSIONS: RL is a good culture media for GNR and D5/W is a poor culture media with the exception of some bacteria of the Tribe Klebsielleae. We recommend to follow high standards of nursing practice for administering intravenous infusions and to avoid nutrient-containing solutions for prolonged parenteral use, when possible.

Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance.

Many different definitions for multidrug-resistant (MDR), extensively drug-resistant (XDR) and pandrug-resistant (PDR) bacteria are being used in the medical literature to characterize the different patterns of resistance found in healthcare-associated, antimicrobial-resistant bacteria. A group of international experts came together through a joint initiative by the European Centre for Disease Prevention and Control (ECDC) and the Centers for Disease Control and Prevention (CDC), to create a standardized international terminology with which to describe acquired resistance profiles in Staphylococcus aureus, Enterococcus spp., Enterobacteriaceae (other than Salmonella and Shigella), Pseudomonas aeruginosa and Acinetobacter spp., all bacteria often responsible for healthcare-associated infections and prone to multidrug resistance. Epidemiologically significant antimicrobial categories were constructed for each bacterium. Lists of antimicrobial categories proposed for antimicrobial susceptibility testing were created using documents and breakpoints from the Clinical Laboratory Standards Institute (CLSI), the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and the United States Food and Drug Administration (FDA). MDR was defined as acquired non-susceptibility to at least one agent in three or more antimicrobial categories, XDR was defined as non-susceptibility to at least one agent in all but two or fewer antimicrobial categories (i.e. bacterial isolates remain susceptible to only one or two categories) and PDR was defined as non-susceptibility to all agents in all antimicrobial categories. To ensure correct application of these definitions, bacterial isolates should be tested against all or nearly all of the antimicrobial agents within the antimicrobial categories and selective reporting and suppression of results should be avoided.

Strategies for antimicrobial susceptibility testing of fastidious aerobic bacteria.

There are several "non-standardized" test methodologies for performing antimicrobial susceptibility tests on clinical isolates of fastidious aerobic organisms. Critical to the interpretation of a susceptibility test (or any clinical laboratory test) is correlation of the results with the clinical status of the patient. If previous extensive studies have not been performed for a given antimicrobial-organism combination using a specific method, the results must be interpreted with discretion. Even when documented correlative data are available, strict quality control measures must be followed to ensure optimal performance of the test system. Ideally, these include testing of quality control organisms of known susceptibilities similar to the test isolate. Such quality control strains of fastidious organisms are not readily available; however, they may be obtained through local health departments. In our laboratory, in order to inform clinicians of the limitations of the results generated from antimicrobial susceptibility testing of fastidious aerobic bacteria using non-standardized (not NCCLS) procedures, we have adopted a mechanism for reporting results of disc tests as "presumptive" (Figure 1). When reporting dilution test results that are derived using methods other than those described by the NCCLS dilution protocol, we indicate the modifications employed in the particular test (Figure 2). Our goals are to attempt to identify unusual resistance that may occur and to generate results that are as accurate, precise, and meaningful as possible, yet we must be aware of the limitations of the procedures with which we are working (Table V). It is only with these understandings that we can be of service to our clinicians and patients.

Ciprofloxacin-resistant Haemophilus influenzae infection in a patient with chronic lung disease.

OBJECTIVE: To report a case of ciprofloxacin-resistant Haemophilus influenzae infection in a patient with chronic lung disease who was exposed to multiple courses of antimicrobial therapy. CASE SUMMARY: The patient suffered recurrent pulmonary infections and developed bronchiectasis as a consequence of longstanding, severe, combined immunodeficiency disease. He had received ciprofloxacin on several occasions for treatment and prophylaxis of recurrent pulmonary infections. On a recent admission his usual H. influenzae isolate, which had been highly susceptible to ciprofloxacin (minimum inhibitory concentration [MIC] < or = 0.06 mg/L) on previous admissions, was resistant to ciprofloxacin and ofloxacin (MIC 8 and 16 mg/L, respectively). The patient responded to treatment with ceftizoxime and was discharged with oral cefixime, which was to be taken for a total of two weeks. DISCUSSION: Rare isolates of H. influenzae resistant to ofloxacin and lomefloxacin have been noted in Europe and Asia; however, none resistant to the fluoroquinolones have been previously reported in the US, and no resistance has been reported to ciprofloxacin. We believe that repetitive, cycling exposure to ciprofloxacin may have induced the resistance that developed in this patient's flora. CONCLUSIONS: Fluoroquinolones may be added to the list of drugs to which H. influenzae have become resistant. Only judicious use of these drugs will preserve their activity against important pathogens in community-acquired infections.