Querying automated antibiotic susceptibility testing instruments: a novel population-based active surveillance method for multidrug-resistant gram-negative bacilli.

OBJECTIVE: To describe the implementation of a population-based surveillance system for multidrug-resistant gram-negative bacilli (MDR-GNB). DESIGN: Population-based active surveillance by the Georgia Emerging Infections Program. SETTING: Metropolitan Atlanta, starting November 2010. PATIENTS: Residents with MDR-GNB isolated from urine or a normally sterile site culture. METHODS: Surveillance was implemented in 3 phases: (1) surveying laboratory antibiotic susceptibility testing practices, (2) piloting surveillance to estimate the proportion of GNB that were MDR, and (3) maintaining ongoing active surveillance for carbapenem-nonsusceptible Enterobacteriaceae and Acinetobacter baumannii using the 2010 Clinical and Laboratory Standards Institute (CLSI) breakpoints. Pilot surveillance required developing and installing queries for GNB on the 3 types of automated testing instruments (ATIs), such as MicroScan, in Atlanta's clinical laboratories. Ongoing surveillance included establishing a process to extract data from ATIs consistently, review charts, manage data, and provide feedback to laboratories. RESULTS: Output from laboratory information systems typically used for surveillance would not reliably capture the CLSI breakpoints, but queries developed for the 3 ATIs did. In November 2010, 0.9% of Enterobacteriaceae isolates and 35.7% of A. baumannii isolates from 21 laboratories were carbapenem nonsusceptible. Over a 5-month period, 82 Enterobacteriaceae and 59 A. baumannii were identified as carbapenem nonsusceptible. CONCLUSIONS: Directly querying ATIs, a novel method of active surveillance for MDR-GNB, proved to be a reliable, sustainable, and accurate method that required moderate initial investment and modest maintenance. Ongoing surveillance is critical to assess the burden of and changes in MDR-GNB to inform prevention efforts.

Impact of changes in Clostridium difficile testing practices on stool rejection policies and C. difficile positivity rates across multiple laboratories in the United States.

We describe the adoption of nucleic acid amplification tests (NAAT) for Clostridium difficile diagnosis and their impact on stool rejection policies and C. difficile positivity rates. Of the laboratories with complete surveys, 51 (43%) reported using NAAT in 2011. Laboratories using NAAT had stricter rejection policies and increased positivity rates.

Effect of nucleic acid amplification testing on population-based incidence rates of Clostridium difficile infection.

Nucleic acid amplification testing (NAAT) is increasingly being adopted for diagnosis of Clostridium difficile infection (CDI). Data from 3 states conducting population-based CDI surveillance showed increases ranging from 43% to 67% in CDI incidence attributable to changing from toxin enzyme immunoassays to NAAT. CDI surveillance requires adjustment for testing methods.

Accuracy of six antimicrobial susceptibility methods for testing linezolid against staphylococci and enterococci.

A challenge panel of enterococci (n = 50) and staphylococci (n = 50), including 17 and 15 isolates that were nonsusceptible to linezolid, respectively, were tested with the Clinical and Laboratory Standards Institute broth microdilution and disk diffusion reference methods. In addition, all 100 isolates were tested in parallel by Etest (AB Biodisk, Solna, Sweden), MicroScan WalkAway (Dade, West Sacramento, CA), BD Phoenix (BD Diagnostic Systems, Sparks, MD), VITEK (bioMérieux, Durham, NC), and VITEK 2 (bioMérieux) by using the manufacturers' protocols. Compared to the results of the broth microdilution method for detecting linezolid-nonsusceptible staphylococci and enterococci, MicroScan results showed the highest category agreement (96.0%). The overall categorical agreement levels for VITEK 2, Etest, Phoenix, disk diffusion, and VITEK were 93.0%, 90.0%, 89.6%, 88.0%, and 85.9%, respectively. The essential agreement levels (results within +/-1 doubling dilution of the MIC determined by the reference method) for MicroScan, Phoenix, VITEK 2, Etest, and VITEK were 99.0%, 95.8%, 92.0%, 92.0%, and 85.9%, respectively. The very major error rates for staphylococci were the highest for VITEK (35.7%), Etest (40.0%), and disk diffusion (53.3%), although the total number of resistant isolates tested was small. The very major error rate for enterococci with VITEK was 20.0%. Three systems (MicroScan, VITEK, and VITEK 2) provided no interpretations of nonsusceptible results for staphylococci. These data, from a challenge panel of isolates, illustrate that the recent emergence of linezolid-nonsusceptible staphylococci and enterococci is providing a challenge for many susceptibility testing systems.

Class 1 integrons in resistant Escherichia coli and Klebsiella spp., US hospitals.

We examined Escherichia coli and Klebsiella spp. from US hospitals for class 1 integrons. Of 320 isolates, 181 (57%) were positive; association of integrons with resistance varied by drug and organism. Thus, determining integron epidemiology will improve understanding of how antibacterial resistance determinants spread in the United States.

Experimental prediction of the evolution of ceftazidime resistance in the CTX-M-2 extended-spectrum beta-lactamase.

We applied in vitro evolution to an Escherichia coli strain containing bla(CTX-M-2) and obtained 10 independent mutant bla(CTX-M-2) alleles that confer elevated resistance to ceftazidime (MIC > or = 32 microg/ml) but lost the ability to confer resistance to cefepime. All alleles had a Pro-to-Ser substitution at position 167.