Evaluation of spa-typing of methicillin-resistant Staphylococcus aureus using high-resolution melting analysis

Waleed Mazi, Vartul Sangal, Gunnar Sandstom, Amir Saeed, Jun Yu

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

OBJECTIVE: spa-typing of methicillin-resistant Staphylococcus aureus (MRSA) has been widely used in clinical diagnostics and epidemiological studies. We aimed to evaluate high-resolution melting (HRM) as a rapid and cost effective method, to replace DNA-sequencing, for spa-typing in a global collection of 50 MRSA isolates.
METHODS: The polymorphic X region of spa gene was amplified by colony PCR using SensiMix™ HRM kit and melting temperature (Tm) and melting curves of the amplicons was analyzed in close tubes using a Rotor-Gene 6000 instrument.
RESULTS: Fifteen out of nineteen spa-types each had distinct Tm, which was sufficient to unambiguously type each of these spa-types. The remaining 4 spa-types cannot be separated by Tm alone: t008 and t2770 shared Tm (80.3°C) and t021 and t311 shared Tm (80.0°C). But, they can be separated based on the shapes of their melting curves. There are discrepancies between ours and previous studies, suggesting that standardization reminds a challenge for cross references.
CONCLUSION: HRM-spa-typing is reproducible, simple, rapid and cost-effective. t037 is prevalent in Brazil and Sudan while diverse spa-types are found in Scotland and Saudi Arabia. Standardization is required for cross-references between labs globally.
LanguageEnglish
Article numberS1201-9712(15)00109-5
Pages1-18
Number of pages18
JournalInternational Journal of Infectious Diseases
Early online date11 May 2015
DOIs
Publication statusPublished - 2015

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Methicillin-Resistant Staphylococcus aureus
Freezing
Temperature
Costs and Cost Analysis
Sudan
Saudi Arabia
Scotland
DNA Sequence Analysis
Genes
Brazil
Epidemiologic Studies
Polymerase Chain Reaction

Keywords

  • high-resolution melting (HRM)
  • spa-types
  • melting temperature (Tm)
  • methicillin resistant Staphylococcus aureus (MRSA)

Cite this

@article{7690d315268348b29ad94c0475465b2e,
title = "Evaluation of spa-typing of methicillin-resistant Staphylococcus aureus using high-resolution melting analysis",
abstract = "OBJECTIVE: spa-typing of methicillin-resistant Staphylococcus aureus (MRSA) has been widely used in clinical diagnostics and epidemiological studies. We aimed to evaluate high-resolution melting (HRM) as a rapid and cost effective method, to replace DNA-sequencing, for spa-typing in a global collection of 50 MRSA isolates.METHODS: The polymorphic X region of spa gene was amplified by colony PCR using SensiMix™ HRM kit and melting temperature (Tm) and melting curves of the amplicons was analyzed in close tubes using a Rotor-Gene 6000 instrument.RESULTS: Fifteen out of nineteen spa-types each had distinct Tm, which was sufficient to unambiguously type each of these spa-types. The remaining 4 spa-types cannot be separated by Tm alone: t008 and t2770 shared Tm (80.3°C) and t021 and t311 shared Tm (80.0°C). But, they can be separated based on the shapes of their melting curves. There are discrepancies between ours and previous studies, suggesting that standardization reminds a challenge for cross references.CONCLUSION: HRM-spa-typing is reproducible, simple, rapid and cost-effective. t037 is prevalent in Brazil and Sudan while diverse spa-types are found in Scotland and Saudi Arabia. Standardization is required for cross-references between labs globally.",
keywords = "high-resolution melting (HRM), spa-types, melting temperature (Tm), methicillin resistant Staphylococcus aureus (MRSA)",
author = "Waleed Mazi and Vartul Sangal and Gunnar Sandstom and Amir Saeed and Jun Yu",
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Evaluation of spa-typing of methicillin-resistant Staphylococcus aureus using high-resolution melting analysis. / Mazi, Waleed; Sangal, Vartul; Sandstom, Gunnar ; Saeed, Amir ; Yu, Jun.

In: International Journal of Infectious Diseases, 2015, p. 1-18.

Research output: Contribution to journalArticle

TY - JOUR

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AU - Saeed, Amir

AU - Yu, Jun

PY - 2015

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N2 - OBJECTIVE: spa-typing of methicillin-resistant Staphylococcus aureus (MRSA) has been widely used in clinical diagnostics and epidemiological studies. We aimed to evaluate high-resolution melting (HRM) as a rapid and cost effective method, to replace DNA-sequencing, for spa-typing in a global collection of 50 MRSA isolates.METHODS: The polymorphic X region of spa gene was amplified by colony PCR using SensiMix™ HRM kit and melting temperature (Tm) and melting curves of the amplicons was analyzed in close tubes using a Rotor-Gene 6000 instrument.RESULTS: Fifteen out of nineteen spa-types each had distinct Tm, which was sufficient to unambiguously type each of these spa-types. The remaining 4 spa-types cannot be separated by Tm alone: t008 and t2770 shared Tm (80.3°C) and t021 and t311 shared Tm (80.0°C). But, they can be separated based on the shapes of their melting curves. There are discrepancies between ours and previous studies, suggesting that standardization reminds a challenge for cross references.CONCLUSION: HRM-spa-typing is reproducible, simple, rapid and cost-effective. t037 is prevalent in Brazil and Sudan while diverse spa-types are found in Scotland and Saudi Arabia. Standardization is required for cross-references between labs globally.

AB - OBJECTIVE: spa-typing of methicillin-resistant Staphylococcus aureus (MRSA) has been widely used in clinical diagnostics and epidemiological studies. We aimed to evaluate high-resolution melting (HRM) as a rapid and cost effective method, to replace DNA-sequencing, for spa-typing in a global collection of 50 MRSA isolates.METHODS: The polymorphic X region of spa gene was amplified by colony PCR using SensiMix™ HRM kit and melting temperature (Tm) and melting curves of the amplicons was analyzed in close tubes using a Rotor-Gene 6000 instrument.RESULTS: Fifteen out of nineteen spa-types each had distinct Tm, which was sufficient to unambiguously type each of these spa-types. The remaining 4 spa-types cannot be separated by Tm alone: t008 and t2770 shared Tm (80.3°C) and t021 and t311 shared Tm (80.0°C). But, they can be separated based on the shapes of their melting curves. There are discrepancies between ours and previous studies, suggesting that standardization reminds a challenge for cross references.CONCLUSION: HRM-spa-typing is reproducible, simple, rapid and cost-effective. t037 is prevalent in Brazil and Sudan while diverse spa-types are found in Scotland and Saudi Arabia. Standardization is required for cross-references between labs globally.

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