Evaluation of high-resolution melting analysis for spa-typing of methicillin-resistant and -susceptible Staphylococcus aureus isolates
Molecular characterization of Staphylococcus aureus isolates from nosocomial and community-acquired infections using accurate, reproducible, and rapid typing methods is essential for the fast identification of prevalent and epidemic strains. Although sequence-based spa typing is highly effective, PC...
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| Format: | Article |
| Language: | English |
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Elsevier
2019-11-01
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| Series: | New Microbes and New Infections |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2052297519301155 |
| _version_ | 1818126478168031232 |
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| author | E. Ohadi S.S. Khoramrooz B.S. Kalani M. Mirzaii R. Pouriran M.R.B. Nasrabadi D. Darban-Sarokhalil |
| author_facet | E. Ohadi S.S. Khoramrooz B.S. Kalani M. Mirzaii R. Pouriran M.R.B. Nasrabadi D. Darban-Sarokhalil |
| author_sort | E. Ohadi |
| collection | DOAJ |
| description | Molecular characterization of Staphylococcus aureus isolates from nosocomial and community-acquired infections using accurate, reproducible, and rapid typing methods is essential for the fast identification of prevalent and epidemic strains. Although sequence-based spa typing is highly effective, PCR-based techniques (such as high-resolution melting curve analysis, HRM) are simpler, less expensive, faster, and can be performed in a single and closed-tube assay format, thereby reducing the risk of contamination. A total of 51 methicillin-resistant S. aureus (MRSA) (n = 26) and methicillin-sensitive S. aureus (MSSA) (n = 25) isolates from Karaj (n = 10) and Yasuj (n = 41), Iran, were subjected to HRM. All selected isolates were identified by the standard spa-typing method. Among the 51 tested isolates, 11 genotype profiles were distinguished from 12 spa types. Strains t1077 and t1816 exhibited the highest and lowest melting temperatures (81.8°C and 79.4°C), with 46.7% and 39.8% G + C contents, respectively. Strains t706 and t1816, with almost identical G + C contents, had the same HRM genotypes, but their curves differed due to different G + C distributions. Four standard spa types (strains t030, t037, t701 and t5598) were differentiated correctly and their melting temperatures were 81.2°C, 81.4°C, 80.4°C and 80.1°C, respectively. We demonstrated that HRM profiling is a rapid method which enables the accurate screening of certain strains (especially the endemic ones), and may be used for bacterial surveillance. However, it cannot replace sequence-based spa typing, especially for newly emerging spa types, and therefore cannot be used as a standardized global method. Keywords: High-resolution melting analysis, HRM, Iran, MRSA, MSSA, spa type |
| first_indexed | 2024-12-11T07:02:06Z |
| format | Article |
| id | doaj.art-cb5c2803055846338ceb4db0e5f89ede |
| institution | Directory Open Access Journal |
| issn | 2052-2975 |
| language | English |
| last_indexed | 2024-12-11T07:02:06Z |
| publishDate | 2019-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | New Microbes and New Infections |
| spelling | doaj.art-cb5c2803055846338ceb4db0e5f89ede2022-12-22T01:16:37ZengElsevierNew Microbes and New Infections2052-29752019-11-0132Evaluation of high-resolution melting analysis for spa-typing of methicillin-resistant and -susceptible Staphylococcus aureus isolatesE. Ohadi0S.S. Khoramrooz1B.S. Kalani2M. Mirzaii3R. Pouriran4M.R.B. Nasrabadi5D. Darban-Sarokhalil6Department of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, IranCellular and Molecular Research Center and Department of Microbiology, Faculty of Medicine, Yasuj University of Medical Sciences, Yasuj, IranDepartment of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, IranSchool of Medicine, Shahroud University of Medical Sciences, Shahroud, IranSchool of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, IranDepartment of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, IranDepartment of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran; Corresponding author: D. Darban-Sarokhalil, Department of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.Molecular characterization of Staphylococcus aureus isolates from nosocomial and community-acquired infections using accurate, reproducible, and rapid typing methods is essential for the fast identification of prevalent and epidemic strains. Although sequence-based spa typing is highly effective, PCR-based techniques (such as high-resolution melting curve analysis, HRM) are simpler, less expensive, faster, and can be performed in a single and closed-tube assay format, thereby reducing the risk of contamination. A total of 51 methicillin-resistant S. aureus (MRSA) (n = 26) and methicillin-sensitive S. aureus (MSSA) (n = 25) isolates from Karaj (n = 10) and Yasuj (n = 41), Iran, were subjected to HRM. All selected isolates were identified by the standard spa-typing method. Among the 51 tested isolates, 11 genotype profiles were distinguished from 12 spa types. Strains t1077 and t1816 exhibited the highest and lowest melting temperatures (81.8°C and 79.4°C), with 46.7% and 39.8% G + C contents, respectively. Strains t706 and t1816, with almost identical G + C contents, had the same HRM genotypes, but their curves differed due to different G + C distributions. Four standard spa types (strains t030, t037, t701 and t5598) were differentiated correctly and their melting temperatures were 81.2°C, 81.4°C, 80.4°C and 80.1°C, respectively. We demonstrated that HRM profiling is a rapid method which enables the accurate screening of certain strains (especially the endemic ones), and may be used for bacterial surveillance. However, it cannot replace sequence-based spa typing, especially for newly emerging spa types, and therefore cannot be used as a standardized global method. Keywords: High-resolution melting analysis, HRM, Iran, MRSA, MSSA, spa typehttp://www.sciencedirect.com/science/article/pii/S2052297519301155 |
| spellingShingle | E. Ohadi S.S. Khoramrooz B.S. Kalani M. Mirzaii R. Pouriran M.R.B. Nasrabadi D. Darban-Sarokhalil Evaluation of high-resolution melting analysis for spa-typing of methicillin-resistant and -susceptible Staphylococcus aureus isolates New Microbes and New Infections |
| title | Evaluation of high-resolution melting analysis for spa-typing of methicillin-resistant and -susceptible Staphylococcus aureus isolates |
| title_full | Evaluation of high-resolution melting analysis for spa-typing of methicillin-resistant and -susceptible Staphylococcus aureus isolates |
| title_fullStr | Evaluation of high-resolution melting analysis for spa-typing of methicillin-resistant and -susceptible Staphylococcus aureus isolates |
| title_full_unstemmed | Evaluation of high-resolution melting analysis for spa-typing of methicillin-resistant and -susceptible Staphylococcus aureus isolates |
| title_short | Evaluation of high-resolution melting analysis for spa-typing of methicillin-resistant and -susceptible Staphylococcus aureus isolates |
| title_sort | evaluation of high resolution melting analysis for spa typing of methicillin resistant and susceptible staphylococcus aureus isolates |
| url | http://www.sciencedirect.com/science/article/pii/S2052297519301155 |
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