Journal of Infectious Diseases & Therapy
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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 sonneil genotyping using HRM analysis for epidemiological purposes. Methods: Fifty clinical MRSA isolates were selected randomly from Scotland, Brazil, Sudan and Saudi Arabia. Methicillinresistant 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 RotorGene 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
Waleed A Mazi 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.
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