Abstracts

Poster Abstracts
uted across the genome, which suggests that
hotspots within the genome may exist where it
will be difficult to accurately detect differences
between two samples. These results provide
critical metrics that show the CFSAN SNP
Pipeline to be a robust method for constructing
a SNP matrix and further reinforces the utility
and importance of validation exercises.
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TGS-TB: TOTAL GENOTYPING SOLUTION
FOR MYCOBACTERIUM TUBERCULOSIS
USING SHORT-READ WHOLE-GENOME
SEQUENCING
T. Sekizuka 1 , A. Yamashita 1 , Y. Murase 2 , T.
Iwamoto 3 , S. Mitarai 2 , S. Kato 2 , M. Kuroda 1 ;
1
National Institute of Infectious Diseases, Shinjyuku-ku,
JAPAN, 2 Japan Anti-Tuberculosis Association,
Kiyose-shi, JAPAN, 3 Kobe Institute
of Health, Kobe-shi, JAPAN.
Background: Whole-genome sequencing
(WGS) with next-generation DNA sequencing
(NGS) is an increasingly accessible and affordable
method for genotyping hundreds of Mycobacterium
tuberculosis (Mtb) isolates, leading
to more effective epidemiological studies
involving single nucleotide variations (SNVs)
in the core genomic sequences based on molecular
evolution. Methods: We developed an
all-in-one web-based tool for genotyping Mtb,
referred to as Total Genotyping Solution for
TB (TGS-TB), to facilitate multiple genotyping
platforms using NGS for spoligotyping and
the detection of phylogenes with core genomic
single nucleotide variations (SNVs), IS6110
insertion sites, and VNTRs (our customized
short TR on 43 loci) through a user-friendly
simple click interface. In addition, this methodology
is implemented with a KvarQ script
to predict MTBC lineages/sublineages and
potential antimicrobial resistance. Findings:
The results of in silico analyses using TGS-TB
are completely consistent with those obtained
using conventional molecular genotyping
methods, suggesting that MiSeq NGS short
reads could provide multiple genotypes to
discriminate multiple strains of Mtb. Indeed,
seven Mtb isolates showing the same VNTR
profile were accurately discriminated through
median joining network analysis using specific
SNVs unique to those isolates. Furthermore,
an additional IS6110 insertion was detected
in one of those isolates as supportive genetic
information in addition to core genomic SNVs.
The results obtained from all in silico analyses
can be downloaded from the website. Interpretation:
TGS-TB provides more accurate
and discriminative strain typing for clinical
and epidemiological investigations; NGS strain
typing offers a total genotyping solution for
Mtb outbreak and surveillance. The genotype
information obtained for all Mtb isolates can
be deposited into an integrated database for
the surveillance of future outbreaks and global
infections. TGS-TB web site: http://gph.
niid.go.jp/tgs-tb Funding: This research was
funded through a Grant-in-Aid for Research
on Emerging and Re-emerging Infectious Diseases
(H25-Shinko-Ippan-015) from the Ministry
of Health Labour and Welfare Programs
of Japan.
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MARA: THE MULTI-ANTIBIOTIC RESISTANCE
ANNOTATOR
S. Partridge 1 , G. Tsafnat 2 ;
1
Westmead Millennium Institute, Sydney, AUS-
TRALIA, 2 Centre for Health Informatics, Macquarie
University, Sydney, AUSTRALIA.
Much of the increasingly problematic multiresistance
in Gram-negative bacteria is due
to resistance genes associated with different
mobile elements (mainly gene cassettes/
integrons, insertion sequences, transposons)
that tend to cluster together in complex multiresistance
regions (MRR). MRR in turn are
found on plasmids that can spread between
cells, including different species, or sometimes
in islands integrated into the chromosome.
Increasing numbers of MRR sequences are
becoming available as part of large projects
using next-generation methods, enabling
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ASM Conferences