Anti-PBP2a (aa 475-498) polyclonal antibody

Purpose. The performance of a rapid test for methicillin-resistant Staphylococcus aureus (MRSA) in a large community hospital was investigated. Methods. A prospective observational study was conducted to evaluate an immunochromatographic assay (Alere PBP2a Culture Colony Test, Alere Scarborough, Inc.) for rapid differentiation of MRSA and methicillin-susceptible S. aureus (MSSA) strains using isolates cultured overnight on common laboratory media. S. aureus isolates cultured for 12-24 hours were tested with the assay, which detects penicillin-binding protein 2a (PBP2a) and provides results in six minutes. The test results were compared with data from standard overnight antimicrobial susceptibility testing to determine the assay's sensitivity and specificity. Changes in therapy associated with use of the rapid assay were evaluated. Results. Over an 11-month period, 661 inpatient isolates from mostly nonhematologic sites were tested. There were six false-negative results, indicating assay sensitivity of 98.4%, with no false positives (specificity of 100%). Eight invalid test results were documented. During designated evaluation periods, a total of 169 patient cases involving PBP2a testing were reviewed by the hospital's antimicrobial stewardship pharmacist. In 63 of those cases (37%), changes in therapy were implemented on the day of test result posting. Interventions often involved switching patients from inappropriate to appropriate MRSA therapy or optimizing MRSA- or MSSA-targeted therapy. Conclusion. An assay for quickly differentiating between MRSA and MSSA was highly sensitive, highly specific, and inexpensive in actual hospital use and led to rapid prescription of appropriate antistaphylococcal therapy 24-48 hours after culture specimens were collected.

Background: Lactic acid bacteria (LAB) are applied worldwide in the production of a variety of fermented food products. Additionally, specific Lactobacillus species are nowadays recognized for their health-promoting effects on the consumer. To optimally exert such beneficial effects, it is considered of great importance that these probiotic bacteria reach their target sites in the gut alive. Methodology/Principal Findings: In the accompanying manuscript by Bron et al. the probiotic model organism Lactobacillus plantarum WCFS1 was cultured under different fermentation conditions, which was complemented by the determination of the corresponding molecular responses by full-genome transcriptome analyses. Here, the gastrointestinal (GI) survival of the cultures produced was assessed in an in vitro assay. Variations in fermentation conditions led to dramatic differences in GI-tract survival (up to 7-log) and high robustness could be associated with low salt and low pH during the fermentations. Moreover, random forest correlation analyses allowed the identification of specific transcripts associated with robustness. Subsequently, the corresponding genes were targeted by genetic engineering, aiming to enhance robustness, which could be achieved for 3 of the genes that negatively correlated with robustness and where deletion derivatives displayed enhanced survival compared to the parental strain. Specifically, a role in GI-tract survival could be confirmed for the lp_1669-encoded AraC-family transcription regulator, involved in capsular polysaccharide remodeling, the penicillin-binding protein Pbp2A involved in peptidoglycan biosynthesis, and the Na+/H+ antiporter NapA3. Moreover, additional physiological analysis established a role for Pbp2A and NapA3 in bile salt and salt tolerance, respectively. Conclusion: Transcriptome trait matching enabled the identification of biomarkers for bacterial (gut-) robustness, which is important for our molecular understanding of GI-tract survival and could facilitate the design of culture conditions aimed to enhance probiotic culture robustness.