5 / 5
Volume 5, Issue 7 (Suppl)
J Infect Dis Ther, an open access journal
ISSN: 2332-0877
Infection Prevention 2017
December 14-15, 2017
Page 32
conference
series
.com
December 14-15, 2017 | Rome, Italy
13
th
World Congress on
INFECTION PREVENTION AND CONTROL
Waleed A Mazi, J Infect Dis Ther 2017, 5:7(Suppl)
DOI: 10.4172/2332-0877-C1-034
Application of high resolution melting to methicillin resistant
Staphlococcus aureus
and
Shigella
sonnei
genotyping for epidemiological purposes
Introduction:
High resolution melting (HRM) analysis has been used in laboratory medicine as acurate, rapid and cost
effective scheme method. Methicillin resistant
Staphylococcus aureus
(MRSA) infections impose huge risk to public health in
healthcare and community settings worldwide.
Shigella sonnei
has been predominantly responsible for dysentary worldwide.
The organism has only one serotype and is genetically homogeneous. We evaluated MRSA
spa
typing and introduced new tools
for
Shigella sonnei
l genotyping using HRM analysis for epidemiological purposes.
Methods:
Fifty clinical MRSA isolates were selected randomly from Scotland, Brazil, Sudan and Saudi Arabia. Methicillin-
resistant phenotype was determined in accordance with BSAC standards using the Vitek 2system. Ten
Shigella sonnei
DNA
samples were provided by Institut Pasteur, France. Primers for the polymorphic X region of the
spa
gene and the six single
nucleotide polymorphisms (SNPs) within
kduD, deoA, emrA, fdX and menF
were amplified by colony PCR using the SensiMix
HRM kit, and the melting temperature (
Tm
) and melting curves of the amplicons were analyzed in close tubes using a Rotor-
Gene 6000 instrument.
Results:
Fifteen
spa
types detected each had a distinct melting temperature (
Tm
) that unambiguously assigned 44 isolates. Both
t008 and t2770, as well as t311 and t021
spa
types, shared the same
Tm
. The first set run separated lineages I, II and III with
distinctive melting curves and the
Tm
of each allele was at least a half degree away from that of other alleles. The second set run
distinguished the sublineages IIIa, IIIb and IIIc with distinctive melting curves.
Conclusion:
HRM analysis is acurate, rapid and cost effective scheme method for identification of MRSA and Shigella sonnie
for epidemiological purposes
Biography
Waleed AMazi is a regional Director for Infection Prevention and Control, King Abdul Aziz Specialist Hospital, Taif – Saudi Arabia. He also worked in Philosophy of Medical
Science, Clinical Microbiology, Karolinska Institutet, Sweden. He has published international articles on prevention of central line –associated bloodstream infection, WHO-
Hand Hygiene implementation program, prevention sharp injuries in healthcare settings and molecular genotyping for epidemiological purposes and participated as a
poster and oral presenters in many international conferences.
wmazi@moh.gov.saWaleed A Mazi
Directorate of Health Affairs, Taif- Kingdom of Saudi Arabia