Improved protocol for rapid identification of certain spa types using high resolution melting curve analysis

PloS One
Benjamin MayerhoferWerner Ruppitsch

Abstract

Methicillin-resistant Staphylococcus aureus is one of the most significant pathogens associated with health care. For efficient surveillance, control and outbreak investigation, S. aureus typing is essential. A high resolution melting curve analysis was developed and evaluated for rapid identification of the most frequent spa types found in an Austrian hospital consortium covering 2,435 beds. Among 557 methicillin-resistant Staphylococcus aureus isolates 38 different spa types were identified by sequence analysis of the hypervariable region X of the protein A gene (spa). Identification of spa types through their characteristic high resolution melting curve profiles was considerably improved by double spiking with genomic DNA from spa type t030 and spa type t003 and allowed unambiguous and fast identification of the ten most frequent spa types t001 (58%), t003 (12%), t190 (9%), t041 (5%), t022 (2%), t032 (2%), t008 (2%), t002 (1%), t5712 (1%) and t2203 (1%), representing 93% of all isolates within this hospital consortium. The performance of the assay was evaluated by testing samples with unknown spa types from the daily routine and by testing three different high resolution melting curve analysis real-time PCR instruments. The ...Continue Reading

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Citations

Nov 23, 2017·Euro Surveillance : Bulletin Européen Sur Les Maladies Transmissibles = European Communicable Disease Bulletin·Werner RuppitschBurkhard Springer
Sep 7, 2017·Journal of Medical Microbiology·Yasser FasihiDavood Kalantar-Neyestanaki
Apr 20, 2021·EFSA Journal·UNKNOWN European Food Safety Authority, UNKNOWN European Centre for Disease Prevention and Control

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Methods Mentioned

BETA
electrophoresis
PCR

Software Mentioned

Ridom StaphType
Ridom Staphserver
LC 480

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