Genome Note Draft genome sequence of reduced teicoplanin-susceptible and vancomycin-heteroresistant methicillin-resistant Staphylococcus aureus from sepsis cases Yamuna Devi Bakthavatchalam a , Balaji Veeraraghavan a, *, Naveen Kumar Devanga Ragupathi a , Priyanka Babu a , Elakkiya Munuswamy a , Thambu David b a Department of Clinical Microbiology, Christian Medical College, Vellore 632004, Tamil Nadu, India b Department of Medicine (Unit II), Christian Medical College, Vellore 632004, India A R T I C L E I N F O Article history: Received 7 October 2016 Received in revised form 7 December 2016 Accepted 8 December 2016 Available online xxx Keywords: MRSA hVISA tcaA A B S T R A C T Here we report the whole-genome shotgun sequence of six methicillin-resistant Staphylococcus aureus (MRSA) showing reduced susceptibility to both vancomycin and teicoplanin. The typical Indian community-acquired MRSA (CA-MRSA) clone ST772-MRSA-V-t657 was the most common genotype (3/ 6; 50%), followed by ST672-MRSA-IV (2/6; 33%) and ST22-MRSA-IV (1/6; 17%). All strains harboured a mutation in the tcaRAB operon, vraSR, graSR and/or rpoB genes, which are frequently mutated determinants in a heteroresistant vancomycin-intermediate S. aureus (hVISA) phenotype. © 2017 Published by Elsevier Ltd on behalf of International Society for Chemotherapy of Infection and Cancer. Methicillin-resistant Staphylococcus aureus (MRSA) causing nosocomial and community-acquired infections is a significant public health threat. Use of vancomycin in the treatment of MRSA infection is complicated by the emergence of vancomycin- heteroresistant, vancomycin-intermediate and vancomycin-resis- tant S. aureus [1]. Here we present the draft genome sequences of six low-level teicoplanin-resistant MRSA that were heteroresistant to vancomycin isolates (VB9V352, VB12268, VB26276, VB23686, VBV169 and VB31683). All six isolates were recovered during 2015 from different patient with MRSA sepsis admitted to Christian Medical College, a 2600-bed tertiary level hospital in Vellore, South India. Except for isolate VB31683, which was a hospital-acquired MRSA, all isolates were found to be community- acquired MRSA. The length of vancomycin/teicoplanin usage in patients ranged from 3 days to 13 days (mean duration 8 days). Isolates were recovered during antimicrobial therapy from patients showing poor response to vancomycin/teicoplanin. Heteroresistant vancomycin-intermediate S. aureus (hVISA) was detected by population analysis profile—area under the curve (PAP-AUC) method. The PAP-AUC profile ratios were between 0.92 and 1.04 with regard to the hVISA (Mu3) isolate. DNA was isolated from pure cultures using a QIAamp 1 DNA Mini Kit (QIAGEN, Hilden, Germany). Whole-genome shotgun sequencing was performed using an Ion Torrent PGM TM System (Life Technologies, Waltham, MA) with 400-bp chemistry. The raw data generated were assembled de novo using SPAdes Genome Assembler v.5.0.0.0 (http://cab.spbu.ru/software/spades/) embed- ded in Torrent suite server v.5.0.4. The genome sequence was annotated using PATRIC, the bacterial bioinformatics database and analysis resource (http://www.patricbrc.org) [2] and the NCBI Prokaryotic Genome Automatic Annotation Pipeline (PGAAP) (http://www.ncbi.nlm.nih.gov/genomes/static/Pipeline.html). Downstream analysis was performed using the Center for Genomic Epidemiology (CGE) server (http://www.cbs.dtu.dk/services) and PATRIC. The resistance gene profile was analysed using ResFinder 2.1 from the CGE server (https://cge.cbs.dtu.dk//services/Res- Finder/). Sequence types (STs) were determined for all of the isolates in the allele order of arcC, aroE, glpF, gmk, pta, tpi and yqiL by comparing the sequences with the S. aureus multilocus sequence typing (MLST) database (http://saureus.mlst.net/). The annotated genome size of the MRSA isolates ranged from ca. 2.7 Mbp to ca. 2.8 Mbp with coverage of 20–58 (Table 1). The number of coding DNA sequences (CDS) per genome ranged * Corresponding author. E-mail address: vbalaji@cmcvellore.ac.in (B. Veeraraghavan). http://dx.doi.org/10.1016/j.jgar.2016.12.008 2213-7165/© 2017 Published by Elsevier Ltd on behalf of International Society for Chemotherapy of Infection and Cancer. Journal of Global Antimicrobial Resistance 8 (2017) 169–171 Contents lists available at ScienceDirect Journal of Global Antimicrobial Resistance journal homepa ge: www.elsev ier.com/locate /jgar