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Objectives: The aim of this study was to determine the prevalence of the colistin-resistance gene (mcr-1) and the antibiotic-susceptibility profile of mcr-1 positive, colistin-resistant isolates in stool specimens of patients attending a tertiary care hospital in Singapore. Methods: 201 diarrheal stool specimens of patients attending the Changi General Hospital between May to August 2017 were collected and screened for the presence of mcr-1 by culture and molecular methods. Antibiotic-susceptibility profile of mcr-1 positive isolates was determined using the polymyxin B and colistin E-tests and the VITEK 2 system. Results: We observed an unexpectedly high prevalence of mcr-1 in patients attending a tertiary care hospital in Singapore, i.e 6.0% and 8.0% estimated by stool culture and direct stool PCR, respectively. The mcr-1 gene was detected predominantly in Escherichia coli. Antibiotic-susceptibility testing on 12 mcr-1 positive Enterobacteriaceae isolates revealed variable susceptibility profiles with no detection of carbapenem-resistant Enterobacteriaceae. Conclusions: This is the first report of the prevalence of human faecal carriage of mcr-1 in Singapore. Our findings highlight the potential risk of mcr-1 spread among our patient cohort. The mcr-1 gene detection combined with the detection of other resistance gene targets of clinical importance is recommended to pre-empt the spread mcr-1 in our patients. Keywords: mcr-1, human faecal carriage, Stool specimen, Colistin resistance, Singapore

Background/Purpose: Screening for vancomycin-resistant enterococci (VRE) by culture takes days to generate results, while polymerase chain reaction (PCR) testing directly from clinical specimens lacks specificity. The aims of this study were to develop a real-time PCR to detect and identify Enterococcus faecium, Enterococcus faecalis, and vanA and vanB genes, and to evaluate the impact of this PCR on test-reporting times when performing it directly from suspect VRE isolates present on screening chromogenic media. Methods: The tetraplex PCR primers were designed to amplify E. faecium, E. faecalis, and vanA and vanB genes, with melt-curve analysis of PCR products. Following analytical and clinical validation of the molecular assay, PCR testing was performed for target colonies present on VRE chromogenic media. PCR results were evaluated against conventional phenotypic identification and susceptibility testing, with the time to result being monitored for both modalities. Results: A total of 519 colonies from clinical specimens were tested concurrently by real-time PCR and phenotypic methods. In all, 223 isolates were identified with phenotypic vancomycin resistance (vanA, n = 108; vanB, n = 105; non-vanA/vanB = 10), with complete agreement between PCR and phenotypic testing for vancomycin-resistant E. faecium and E. faecalis. The majority (88.6%) of PCR results were reported, on average, 24.8 hours earlier than those of phenotypic testing, with 68% reduction in total costs. Conclusion: The use of culture on selective media, followed by direct colony PCR confirmation allows faster and economical VRE screening.