Anti-VIM monoclonal antibody, clone W0 [Biotin]

Background: The global spread of carbapenemase-producing organisms (CPO) has been considered by international health authorities as a critical public health concern. Brazil has a high CPO prevalence according to distinct publications but many routine microbiology laboratories have only phenotypic resources to evaluate this epidemiological situation, which is time-consuming and detects only carbapenem-resistant isolates missing CPO susceptible expressing a slightly decreased susceptibility. New molecular platforms can detect CPO faster but a local evaluation is essential. Aim: To evaluate the performance of CPO detection direct from rectal swabs with the Xpert Carba-R (TM) assay (Cepheid, Sunnyvale, CA) in the largest Brazilian University Hospital. Methods: A prospective diagnostic accuracy study of CPO was performed with the collection of rectal swabs from patients admitted into the Intensive Care Unit (ICU) and into the Emergency Department (ED) between April and July 2016. The Xpert Carba-R assay results were compared with carbapenem-resistant Enterobacterales (CRE) surveillance cultures plus in-house PCR carbapenemase detection (reference method). In case of discordant results between methods, additional tests were performed. The limit of detection (LoD) for the CRE culture and the Xpert Carba-R (TM) assay were performed with contrived isolates of known carbapenemases genes. Results: A total of 921 clinical rectal swabs were analyzed being 21% (196/921) from the ICU and 79% (725/921) from the ED. Overall, the Xpert Carba-R (TM) assay detected 9.9% (91/921) of CPOs being 9.5% (87/921) positive only for bla(KPC) and 0.4% (4/921) positive only for bla(NDM). The reference method detected 9.1% (84/921) CPO being 77 (8.4%) bla(KPC), 5 bla(VIM) (0.5%) and 2 bla(NDM) (0.2%). No IMP or OXA-48 like gene was detected. Overall, twelve samples, 1.3% (10 bla(KPC), 2 bla(NDM)) were Xpert Carba-R (TM) positive but negative by the reference method. Five isolates (0.5%) were positive for bla(VIM) only by in-house PCR and confirmed to be bla(VIM-2) by DNA sequencing. The Kappa value, sensitivity, specificity, positive/negative predictive values and accuracy of the Xpert Carba-R (TM) assay were; 0.893 (95% confidence interval [CI], 0.842-0.944), 94% (86.7-98.0), 98.6% (97.5-99.3), 86.8% (78.1-93.0), 99.4% (98.6-99.8) and 98.2% (97.3-99.1), respectively. The LoD for bla(KPC) of the Xpert Carba-R (TM) assay and the CRE cultures were 10(1) CFU/swab. Conclusion: The Xpert Carba-R (TM) assay is an accurate test to detect CPO directly from the rectal swabs with significant lower turnaround time (TAT) when compared to the reference method (CRE culture plus in-house PCR). Xpert Carba-R (TM) may, therefore, be regarded as a good and fast epidemiological tool.

The presence of extended-spectrum beta-lactamase (ESBL)-producing bacteria in environmental sources has been reported worldwide and constitutes a serious risk of community-acquired infections with limited treatment options. The current study aimed to explore the presence of these worrisome bacteria in a pond located at the Norwegian University of Life Sciences in angstrom s, Norway. A total of 98 bacterial isolates survived growth on selective chromogenic media and were identified by 16S rRNA Sanger sequencing. All strains were evaluated for the presence of the most commonly found beta-lactamases and ESBLs in clinical settings (bla(CTX-M) groups 1, 2, and 9, bla(CMY), bla(SHV) and bla(TEM)) and carbapenemases (bla(IMP), bla(KPC), bla(NDM), bla(OXA), bla(SFC1), bla(VIM)) through multiplex PCR. A total of eight strains were determined to contain one or more genes of interest. Phenotypic resistance to 18 antimicrobial agents was assessed and isolates were subjected to whole genome sequencing through a combination of Oxford Nanopore's MinION and lllumina's MiSeq. Results revealed the presence of beta-lactamase and ESBL-producing Escherichia coli, Klebsiella pneumoniae, Stenotrophomonas maltophilia, and a Paraburkholderia spp. Identified beta-lactamases and ESBLs include bla(CTX-M), bla(TEM), bla(CMY), bla(SHV) and a possible bla(KPC)-like gene, with both documented and novel sequences established. In addition, two inducible beta-lactamases were found, a class A beta-lactamase (L1) and a cephalosporinase (L2). All strains were determined to be multidrug resistant and numerous resistance genes to non-beta-lactams were observed. In conclusion, this study demonstrates that environmental sources are a potential reservoir of clinically relevant ESBL-producing bacteria that may pose a health risk to humans upon exposure.