Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance

Staphylococcal infection is one of the most pressing problems in modern medicine due to the increasing antibiotic resistance with the overuse of antibiotics. Conventional methods for identification and antimicrobial susceptibility testing (AST) generally take 3–7 days and require skilled technicians...
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Autor*in:	Xiangrui Meng [verfasserIn] Guohao Zhang [verfasserIn] Bo Sun [verfasserIn] Shujun Liu [verfasserIn] Yadong Wang [verfasserIn] Ming Gao [verfasserIn] Yubo Fan [verfasserIn] Guojun Zhang [verfasserIn] Guangzhi Shi [verfasserIn] Xixiong Kang [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2020

Schlagwörter:	microfluidics staphylococcal infection loop-mediated isothermal amplification identification methicillin resistance

Übergeordnetes Werk:	In: Frontiers in Microbiology - Frontiers Media S.A., 2011, 11(2020)
Übergeordnetes Werk:	volume:11 ; year:2020

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3389/fmicb.2020.01487

Katalog-ID:	DOAJ045533024

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allfields	10.3389/fmicb.2020.01487 doi (DE-627)DOAJ045533024 (DE-599)DOAJfea1b7b4f8da4cc19d8355e9a258fe3b DE-627 ger DE-627 rakwb eng QR1-502 Xiangrui Meng verfasserin aut Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Staphylococcal infection is one of the most pressing problems in modern medicine due to the increasing antibiotic resistance with the overuse of antibiotics. Conventional methods for identification and antimicrobial susceptibility testing (AST) generally take 3–7 days and require skilled technicians. In this study, a microfluidic device based on loop-mediated isothermal amplification (LAMP) was developed, which could discriminate Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis and predict their methicillin resistance by targeting the mecA and femA genes within 70 min including the hands-on time. Multiplex and real-time detection was achieved in a closed system without aerosol contamination. The limits of detection (LODs) for S. aureus, S. epidermidis, S. hominis, and methicillin-resistant S. aureus (MRSA) were 20 CFU/reaction, while that for S. haemolyticus was 200 CFU/reaction. A total of 102 positive cultures of cerebrospinal fluid (CSF) were also tested, and the results were in good agreement with those from conventional methods. Furthermore, mixed cultures were readily identified by our method. The portable and integrated device is rapid, accurate, and easy to use, which can provide information for prompt institution of proper antimicrobial therapy and has great potential for clinical applications, especially in resource-limited settings. microfluidics staphylococcal infection loop-mediated isothermal amplification identification methicillin resistance Microbiology Xiangrui Meng verfasserin aut Guohao Zhang verfasserin aut Bo Sun verfasserin aut Shujun Liu verfasserin aut Shujun Liu verfasserin aut Yadong Wang verfasserin aut Ming Gao verfasserin aut Ming Gao verfasserin aut Yubo Fan verfasserin aut Yubo Fan verfasserin aut Guojun Zhang verfasserin aut Guojun Zhang verfasserin aut Guangzhi Shi verfasserin aut Xixiong Kang verfasserin aut Xixiong Kang verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 11(2020) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:11 year:2020 https://doi.org/10.3389/fmicb.2020.01487 kostenfrei https://doaj.org/article/fea1b7b4f8da4cc19d8355e9a258fe3b kostenfrei https://www.frontiersin.org/article/10.3389/fmicb.2020.01487/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2020
spelling	10.3389/fmicb.2020.01487 doi (DE-627)DOAJ045533024 (DE-599)DOAJfea1b7b4f8da4cc19d8355e9a258fe3b DE-627 ger DE-627 rakwb eng QR1-502 Xiangrui Meng verfasserin aut Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Staphylococcal infection is one of the most pressing problems in modern medicine due to the increasing antibiotic resistance with the overuse of antibiotics. Conventional methods for identification and antimicrobial susceptibility testing (AST) generally take 3–7 days and require skilled technicians. In this study, a microfluidic device based on loop-mediated isothermal amplification (LAMP) was developed, which could discriminate Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis and predict their methicillin resistance by targeting the mecA and femA genes within 70 min including the hands-on time. Multiplex and real-time detection was achieved in a closed system without aerosol contamination. The limits of detection (LODs) for S. aureus, S. epidermidis, S. hominis, and methicillin-resistant S. aureus (MRSA) were 20 CFU/reaction, while that for S. haemolyticus was 200 CFU/reaction. A total of 102 positive cultures of cerebrospinal fluid (CSF) were also tested, and the results were in good agreement with those from conventional methods. Furthermore, mixed cultures were readily identified by our method. The portable and integrated device is rapid, accurate, and easy to use, which can provide information for prompt institution of proper antimicrobial therapy and has great potential for clinical applications, especially in resource-limited settings. microfluidics staphylococcal infection loop-mediated isothermal amplification identification methicillin resistance Microbiology Xiangrui Meng verfasserin aut Guohao Zhang verfasserin aut Bo Sun verfasserin aut Shujun Liu verfasserin aut Shujun Liu verfasserin aut Yadong Wang verfasserin aut Ming Gao verfasserin aut Ming Gao verfasserin aut Yubo Fan verfasserin aut Yubo Fan verfasserin aut Guojun Zhang verfasserin aut Guojun Zhang verfasserin aut Guangzhi Shi verfasserin aut Xixiong Kang verfasserin aut Xixiong Kang verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 11(2020) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:11 year:2020 https://doi.org/10.3389/fmicb.2020.01487 kostenfrei https://doaj.org/article/fea1b7b4f8da4cc19d8355e9a258fe3b kostenfrei https://www.frontiersin.org/article/10.3389/fmicb.2020.01487/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2020
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allfieldsGer	10.3389/fmicb.2020.01487 doi (DE-627)DOAJ045533024 (DE-599)DOAJfea1b7b4f8da4cc19d8355e9a258fe3b DE-627 ger DE-627 rakwb eng QR1-502 Xiangrui Meng verfasserin aut Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Staphylococcal infection is one of the most pressing problems in modern medicine due to the increasing antibiotic resistance with the overuse of antibiotics. Conventional methods for identification and antimicrobial susceptibility testing (AST) generally take 3–7 days and require skilled technicians. In this study, a microfluidic device based on loop-mediated isothermal amplification (LAMP) was developed, which could discriminate Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis and predict their methicillin resistance by targeting the mecA and femA genes within 70 min including the hands-on time. Multiplex and real-time detection was achieved in a closed system without aerosol contamination. The limits of detection (LODs) for S. aureus, S. epidermidis, S. hominis, and methicillin-resistant S. aureus (MRSA) were 20 CFU/reaction, while that for S. haemolyticus was 200 CFU/reaction. A total of 102 positive cultures of cerebrospinal fluid (CSF) were also tested, and the results were in good agreement with those from conventional methods. Furthermore, mixed cultures were readily identified by our method. The portable and integrated device is rapid, accurate, and easy to use, which can provide information for prompt institution of proper antimicrobial therapy and has great potential for clinical applications, especially in resource-limited settings. microfluidics staphylococcal infection loop-mediated isothermal amplification identification methicillin resistance Microbiology Xiangrui Meng verfasserin aut Guohao Zhang verfasserin aut Bo Sun verfasserin aut Shujun Liu verfasserin aut Shujun Liu verfasserin aut Yadong Wang verfasserin aut Ming Gao verfasserin aut Ming Gao verfasserin aut Yubo Fan verfasserin aut Yubo Fan verfasserin aut Guojun Zhang verfasserin aut Guojun Zhang verfasserin aut Guangzhi Shi verfasserin aut Xixiong Kang verfasserin aut Xixiong Kang verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 11(2020) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:11 year:2020 https://doi.org/10.3389/fmicb.2020.01487 kostenfrei https://doaj.org/article/fea1b7b4f8da4cc19d8355e9a258fe3b kostenfrei https://www.frontiersin.org/article/10.3389/fmicb.2020.01487/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2020
allfieldsSound	10.3389/fmicb.2020.01487 doi (DE-627)DOAJ045533024 (DE-599)DOAJfea1b7b4f8da4cc19d8355e9a258fe3b DE-627 ger DE-627 rakwb eng QR1-502 Xiangrui Meng verfasserin aut Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Staphylococcal infection is one of the most pressing problems in modern medicine due to the increasing antibiotic resistance with the overuse of antibiotics. Conventional methods for identification and antimicrobial susceptibility testing (AST) generally take 3–7 days and require skilled technicians. In this study, a microfluidic device based on loop-mediated isothermal amplification (LAMP) was developed, which could discriminate Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis and predict their methicillin resistance by targeting the mecA and femA genes within 70 min including the hands-on time. Multiplex and real-time detection was achieved in a closed system without aerosol contamination. The limits of detection (LODs) for S. aureus, S. epidermidis, S. hominis, and methicillin-resistant S. aureus (MRSA) were 20 CFU/reaction, while that for S. haemolyticus was 200 CFU/reaction. A total of 102 positive cultures of cerebrospinal fluid (CSF) were also tested, and the results were in good agreement with those from conventional methods. Furthermore, mixed cultures were readily identified by our method. The portable and integrated device is rapid, accurate, and easy to use, which can provide information for prompt institution of proper antimicrobial therapy and has great potential for clinical applications, especially in resource-limited settings. microfluidics staphylococcal infection loop-mediated isothermal amplification identification methicillin resistance Microbiology Xiangrui Meng verfasserin aut Guohao Zhang verfasserin aut Bo Sun verfasserin aut Shujun Liu verfasserin aut Shujun Liu verfasserin aut Yadong Wang verfasserin aut Ming Gao verfasserin aut Ming Gao verfasserin aut Yubo Fan verfasserin aut Yubo Fan verfasserin aut Guojun Zhang verfasserin aut Guojun Zhang verfasserin aut Guangzhi Shi verfasserin aut Xixiong Kang verfasserin aut Xixiong Kang verfasserin aut In Frontiers in Microbiology Frontiers Media S.A., 2011 11(2020) (DE-627)642889384 (DE-600)2587354-4 1664302X nnns volume:11 year:2020 https://doi.org/10.3389/fmicb.2020.01487 kostenfrei https://doaj.org/article/fea1b7b4f8da4cc19d8355e9a258fe3b kostenfrei https://www.frontiersin.org/article/10.3389/fmicb.2020.01487/full kostenfrei https://doaj.org/toc/1664-302X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2020
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callnumber-first	Q - Science
author	Xiangrui Meng
spellingShingle	Xiangrui Meng misc QR1-502 misc microfluidics misc staphylococcal infection misc loop-mediated isothermal amplification misc identification misc methicillin resistance misc Microbiology Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance
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topic_title	QR1-502 Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance microfluidics staphylococcal infection loop-mediated isothermal amplification identification methicillin resistance
topic	misc QR1-502 misc microfluidics misc staphylococcal infection misc loop-mediated isothermal amplification misc identification misc methicillin resistance misc Microbiology
topic_unstemmed	misc QR1-502 misc microfluidics misc staphylococcal infection misc loop-mediated isothermal amplification misc identification misc methicillin resistance misc Microbiology
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title	Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance
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title_full	Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance
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title_sort	rapid detection of meca and fema genes by loop-mediated isothermal amplification in a microfluidic system for discrimination of different staphylococcal species and prediction of methicillin resistance
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title_auth	Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance
abstract	Staphylococcal infection is one of the most pressing problems in modern medicine due to the increasing antibiotic resistance with the overuse of antibiotics. Conventional methods for identification and antimicrobial susceptibility testing (AST) generally take 3–7 days and require skilled technicians. In this study, a microfluidic device based on loop-mediated isothermal amplification (LAMP) was developed, which could discriminate Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis and predict their methicillin resistance by targeting the mecA and femA genes within 70 min including the hands-on time. Multiplex and real-time detection was achieved in a closed system without aerosol contamination. The limits of detection (LODs) for S. aureus, S. epidermidis, S. hominis, and methicillin-resistant S. aureus (MRSA) were 20 CFU/reaction, while that for S. haemolyticus was 200 CFU/reaction. A total of 102 positive cultures of cerebrospinal fluid (CSF) were also tested, and the results were in good agreement with those from conventional methods. Furthermore, mixed cultures were readily identified by our method. The portable and integrated device is rapid, accurate, and easy to use, which can provide information for prompt institution of proper antimicrobial therapy and has great potential for clinical applications, especially in resource-limited settings.
abstractGer	Staphylococcal infection is one of the most pressing problems in modern medicine due to the increasing antibiotic resistance with the overuse of antibiotics. Conventional methods for identification and antimicrobial susceptibility testing (AST) generally take 3–7 days and require skilled technicians. In this study, a microfluidic device based on loop-mediated isothermal amplification (LAMP) was developed, which could discriminate Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis and predict their methicillin resistance by targeting the mecA and femA genes within 70 min including the hands-on time. Multiplex and real-time detection was achieved in a closed system without aerosol contamination. The limits of detection (LODs) for S. aureus, S. epidermidis, S. hominis, and methicillin-resistant S. aureus (MRSA) were 20 CFU/reaction, while that for S. haemolyticus was 200 CFU/reaction. A total of 102 positive cultures of cerebrospinal fluid (CSF) were also tested, and the results were in good agreement with those from conventional methods. Furthermore, mixed cultures were readily identified by our method. The portable and integrated device is rapid, accurate, and easy to use, which can provide information for prompt institution of proper antimicrobial therapy and has great potential for clinical applications, especially in resource-limited settings.
abstract_unstemmed	Staphylococcal infection is one of the most pressing problems in modern medicine due to the increasing antibiotic resistance with the overuse of antibiotics. Conventional methods for identification and antimicrobial susceptibility testing (AST) generally take 3–7 days and require skilled technicians. In this study, a microfluidic device based on loop-mediated isothermal amplification (LAMP) was developed, which could discriminate Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis and predict their methicillin resistance by targeting the mecA and femA genes within 70 min including the hands-on time. Multiplex and real-time detection was achieved in a closed system without aerosol contamination. The limits of detection (LODs) for S. aureus, S. epidermidis, S. hominis, and methicillin-resistant S. aureus (MRSA) were 20 CFU/reaction, while that for S. haemolyticus was 200 CFU/reaction. A total of 102 positive cultures of cerebrospinal fluid (CSF) were also tested, and the results were in good agreement with those from conventional methods. Furthermore, mixed cultures were readily identified by our method. The portable and integrated device is rapid, accurate, and easy to use, which can provide information for prompt institution of proper antimicrobial therapy and has great potential for clinical applications, especially in resource-limited settings.
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title_short	Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance
url	https://doi.org/10.3389/fmicb.2020.01487 https://doaj.org/article/fea1b7b4f8da4cc19d8355e9a258fe3b https://www.frontiersin.org/article/10.3389/fmicb.2020.01487/full https://doaj.org/toc/1664-302X
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author2	Xiangrui Meng Guohao Zhang Bo Sun Shujun Liu Yadong Wang Ming Gao Yubo Fan Guojun Zhang Guangzhi Shi Xixiong Kang
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Conventional methods for identification and antimicrobial susceptibility testing (AST) generally take 3–7 days and require skilled technicians. In this study, a microfluidic device based on loop-mediated isothermal amplification (LAMP) was developed, which could discriminate Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis and predict their methicillin resistance by targeting the mecA and femA genes within 70 min including the hands-on time. Multiplex and real-time detection was achieved in a closed system without aerosol contamination. The limits of detection (LODs) for S. aureus, S. epidermidis, S. hominis, and methicillin-resistant S. aureus (MRSA) were 20 CFU/reaction, while that for S. haemolyticus was 200 CFU/reaction. A total of 102 positive cultures of cerebrospinal fluid (CSF) were also tested, and the results were in good agreement with those from conventional methods. 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Rapid Detection of mecA and femA Genes by Loop-Mediated Isothermal Amplification in a Microfluidic System for Discrimination of Different Staphylococcal Species and Prediction of Methicillin Resistance

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