Diagnostic performance of the Xpert Carba-R (TM) assay directly from rectal swabs for active surveillance of carbapenemase-producing organisms in the largest Brazilian University Hospital

Abstract

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.