Copyright © 2015 Beck, Eric T. Eric T. Beck 1, Garrett C. Reymann 1, Blake W. Buchan 1,2, and Nathan A. Ledeboer 1,2 1 Department of Microbiology, Dynacare Laboratories, Milwaukee, WI 53226, USA; 2 Department of Pathology, Medical College of Wisconsin, Milwaukee, WI 53226, USA INTRODUCTION/PURPOSE: US Centers for Disease Control guidelines indicate that molecular assays for group B streptococci (GBS) be performed on enriched culture specimens, which negates the rapid turnaround time of these assays. The purpose of this study was to evaluate whether specimens collected with ESwab collection devices ([ESwab] regular flocked swab with 1 mL liquid Amies transport medium; Copan Diagnostics Inc, Murrieta, CA) can be used for directl testing in the BD MAX™ GBS Assay ([MAX] BD Diagnostics, Quebec, Canada). In this study, mock specimens were prepared to determine the optimal input volume of liquid Amies transport medium into the MAX™ GBS Assay. In addition, studies were performed to determine whether Eswabs yield equivalent sensitivity to enriched culture specimens in the MAX™ GBS Assay. Sensitivity of the BD MAX group B Streptococcus assay increases with direct testing of specimens collected in Copan ESwab collection devices vs. enriched culture specimens. CONCLUSIONS:  Routine GBS screening is performed at weeks gestational age, but testing closer to time of delivery would likely increase the negative predictive value of the result.  In order to perform GBS testing at time of admission must have access to a faster, more sensitive GBS testing methodology.  The current study shows that direct testing of Eswabs (without broth enrichment) would decrease the turnaround time of the MAX™ GBS Assay by approximately 90% (24 to 2 hours).  In addition to decreasing TAT, Eswabs increased the sensitivity over 100 fold compared to the current methodology.  This study was performed with mock specimens, but the results merit further clinical investigation. METHODS:  Pool LIM broth cultures inoculated with vaginal-rectal swabs that showed growth, but were negative for the presence of GBS..  Prepare 0.5 McFarland preparation of GBS.  Dilute GBS to 1.5 x 10 6 – 1.5 x 10 1 CFU/mL in the pooled GBS negative cultures.  Prepare mock specimens by dipping Eswabs or standard collection swabs (BBL™ CultureSwab™ Dual Swab with Liquid Stuart Medium; BD Diagnostics, Sparks, MD) into dilutions.  Test different volumes and concentrations of Eswab specimen in the MAX™ GBS Assay to determine optimal input volume.  Prepare mock specimens with four standard collection devices and four ESwabs for each dilution.  Inoculate standard swabs into sterile Lim broth, incubate overnight at 35°C, and test in the MAX assay according to the package insert.  Vortex ESwabs and add 500 µL of transport medium to the sample buffer tube and test immediately in the MAX assay. RESULTS: 500 µL of Eswab transport medium appears to be the optimal input volume for the MAX™ GBS Assay. The limit of detection (the least concentrated dilution in which at least 3 of 4 replicates were positive) of the MAX™ GBS with ESwabs and Lim Broth cultures was 1.5 x 10 2 and 1.5 x 10 4 CFU/mL, respectively. 15 µL Over Night Lim Broth Culture input into MAX GBS Assay Replicate 1.5 x 10 6 CFU/mL 1.5 x 10 5 CFU/mL 1.5 x 10 4 CFU/mL 1.5 x 10 3 CFU/mL 1.5 x 10 2 CFU/mL 1.5 x 10 1 CFU/mL 1POS NEG 2POS NEG 3POS NEG 4POS NEG Flocked Swab in ESwab Medium with 500 µL input into MAX GBS Assay Replicate 1.5 x 10 6 CFU/mL 1.5 x 10 5 CFU/mL 1.5 x 10 4 CFU/mL 1.5 x 10 3 CFU/mL 1.5 x 10 2 CFU/mL 1.5 x 10 1 CFU/mL 1 POS NEG 2 POS NEG 3 POS NEG 4 POS NEG Materials and Support for this study provided by Becton Dickinson and Company GBS Assay Ct Value by Concentration and Eswab Input Volume in MAX GBS Assay ESwab Input Volume 1.5 x 10 6 CFU/mL GBS Ct 1.5 x 10 5 CFU/mL GBS Ct 1.5 x 10 4 CFU/mL GBS Ct 1.5 x 10 3 CFU/mL GBS Ct 1.5 x 1 2 CFU/mL GBS Ct 1.5 x 10 1 CFU/mL GBS Ct 15 µL NEG 50 µL NEG 100 µL NEG 200 µL NEG 300 µL NEG 400 µL NEG 500 µL NEG 900 µL NEG Prepare 0.5 McF of GBS Dilute from in GBS Neg O/N Lim 35° C for 24 hours 500 µL 15 µL