Rapid detection of methicillin-resistant Staphylococcus aureus in positive blood-cultures by recombinase polymerase amplification combined with lateral flow strip.
Staphylococcus aureus, especially methicillin-resistant S. aureus (MRSA), is an important bacterium that causes community and healthcare-related infections throughout the world. However, the current conventional detection methods are time-consuming. We therefore developed and evaluated a recombinase...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2022-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0270686 |
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| author | Arpasiri Srisrattakarn Pimchanok Panpru Patcharaporn Tippayawat Aroonwadee Chanawong Ratree Tavichakorntrakool Jureerut Daduang Lumyai Wonglakorn Aroonlug Lulitanond |
| author_facet | Arpasiri Srisrattakarn Pimchanok Panpru Patcharaporn Tippayawat Aroonwadee Chanawong Ratree Tavichakorntrakool Jureerut Daduang Lumyai Wonglakorn Aroonlug Lulitanond |
| author_sort | Arpasiri Srisrattakarn |
| collection | DOAJ |
| description | Staphylococcus aureus, especially methicillin-resistant S. aureus (MRSA), is an important bacterium that causes community and healthcare-related infections throughout the world. However, the current conventional detection methods are time-consuming. We therefore developed and evaluated a recombinase polymerase amplification-lateral flow strip (RPA-LF) approach for detection of MRSA in positive blood-culture samples. Sixty positive blood-cultures from a hospital were tested directly without DNA extraction and purification before the amplification reaction. RPA primers and probes were designed for nuc (encoding thermonuclease) and mecA (encoding penicillin-binding protein 2a) genes to diagnose S. aureus and its methicillin-resistance status. The RPA reaction occurred under isothermal conditions (45°C) within 20 min and a result was provided by the LF strip in a further 5 min at room temperature. The evaluation of RPA-LF using blood-culture samples showed 93.3% (14/15) sensitivity for identifying S. aureus, and no cross-amplification was seen [100% (45/45) specificity]. For detection of methicillin resistance, the RPA-LF test provided 100% (16/16) sensitivity and 97.7% (43/44) specificity. The RPA-LF is rapid, highly sensitive, robust and easy to use. It can be used for direct detection of MRSA with no requirement for special equipment. |
| first_indexed | 2024-12-10T10:15:36Z |
| format | Article |
| id | doaj.art-29eda6b0dc3e45bbafe3952e6316da95 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-10T10:15:36Z |
| publishDate | 2022-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-29eda6b0dc3e45bbafe3952e6316da952022-12-22T01:53:00ZengPublic Library of Science (PLoS)PLoS ONE1932-62032022-01-01176e027068610.1371/journal.pone.0270686Rapid detection of methicillin-resistant Staphylococcus aureus in positive blood-cultures by recombinase polymerase amplification combined with lateral flow strip.Arpasiri SrisrattakarnPimchanok PanpruPatcharaporn TippayawatAroonwadee ChanawongRatree TavichakorntrakoolJureerut DaduangLumyai WonglakornAroonlug LulitanondStaphylococcus aureus, especially methicillin-resistant S. aureus (MRSA), is an important bacterium that causes community and healthcare-related infections throughout the world. However, the current conventional detection methods are time-consuming. We therefore developed and evaluated a recombinase polymerase amplification-lateral flow strip (RPA-LF) approach for detection of MRSA in positive blood-culture samples. Sixty positive blood-cultures from a hospital were tested directly without DNA extraction and purification before the amplification reaction. RPA primers and probes were designed for nuc (encoding thermonuclease) and mecA (encoding penicillin-binding protein 2a) genes to diagnose S. aureus and its methicillin-resistance status. The RPA reaction occurred under isothermal conditions (45°C) within 20 min and a result was provided by the LF strip in a further 5 min at room temperature. The evaluation of RPA-LF using blood-culture samples showed 93.3% (14/15) sensitivity for identifying S. aureus, and no cross-amplification was seen [100% (45/45) specificity]. For detection of methicillin resistance, the RPA-LF test provided 100% (16/16) sensitivity and 97.7% (43/44) specificity. The RPA-LF is rapid, highly sensitive, robust and easy to use. It can be used for direct detection of MRSA with no requirement for special equipment.https://doi.org/10.1371/journal.pone.0270686 |
| spellingShingle | Arpasiri Srisrattakarn Pimchanok Panpru Patcharaporn Tippayawat Aroonwadee Chanawong Ratree Tavichakorntrakool Jureerut Daduang Lumyai Wonglakorn Aroonlug Lulitanond Rapid detection of methicillin-resistant Staphylococcus aureus in positive blood-cultures by recombinase polymerase amplification combined with lateral flow strip. PLoS ONE |
| title | Rapid detection of methicillin-resistant Staphylococcus aureus in positive blood-cultures by recombinase polymerase amplification combined with lateral flow strip. |
| title_full | Rapid detection of methicillin-resistant Staphylococcus aureus in positive blood-cultures by recombinase polymerase amplification combined with lateral flow strip. |
| title_fullStr | Rapid detection of methicillin-resistant Staphylococcus aureus in positive blood-cultures by recombinase polymerase amplification combined with lateral flow strip. |
| title_full_unstemmed | Rapid detection of methicillin-resistant Staphylococcus aureus in positive blood-cultures by recombinase polymerase amplification combined with lateral flow strip. |
| title_short | Rapid detection of methicillin-resistant Staphylococcus aureus in positive blood-cultures by recombinase polymerase amplification combined with lateral flow strip. |
| title_sort | rapid detection of methicillin resistant staphylococcus aureus in positive blood cultures by recombinase polymerase amplification combined with lateral flow strip |
| url | https://doi.org/10.1371/journal.pone.0270686 |
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