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The rapid and visual detection of methicillin-susceptible and methicillin-resistant Staphylococcus aureus using multiplex loop-mediated isothermal amplification linked to a nanoparticle-based lateral flow biosensor.
Antimicrob Resist Infect Control. 2020 07 17; 9(1):111.AR

Abstract

BACKGROUND

Staphylococcus aureus (S. aureus), including methicillin-susceptible S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA), is an eminent human pathogen that can colonize the human host and cause severe life-threatening infections. The development of a reliable, simple and rapid assay for detecting S. aureus and identifying MRSA is important for diagnosis and follow-up treatment.

METHODS

A novel molecular diagnosis technique, named multiplex loop-mediated isothermal amplification linked to a nanoparticle-based lateral flow biosensor (m-LAMP-LFB), was applied to detect all S. aureus species and identify MRSA. Two sets of primers were designed based on the femA gene (S. aureus-specific gene) and the mecA gene (encoding penicillin-binding protein 2a), and the multiple-LAMP products were analyzed using LFB. The m-LAMP-LFB amplification conditions, including the target DNA concentration, reaction temperature and time, were optimized. The sensitivity and specificity of the m-LAMP-LFB method were tested in the current study, and the multiple-LAMP-LFB technology was applied to detect the MSSA and MRSA strains from clinical samples.

RESULTS

The S. aureus- and MRSA-specific primers based on the femA and mecA genes allowed the multiple-LAMP technology to detect S. aureus and MRSA, respectively. The multiple-LAMP conditions were optimized at 63 °C for 40 min. The full process, including genomic DNA template preparation, LAMP, and product identification, could be achieved in 80 min. The limit of detection (LoD) of the multiple-LAMP assay for femA and mecA detection was 100 fg of genomic DNA template per reaction. The specificity of m-LAMP-LFB detection was 100 %, and no cross-reactions to non-S. aureus strains were observed.

CONCLUSION

The multiple-LAMP-LFB technique developed in the current study is a reliable, simple, rapid, specific and sensitive method to identify MSSA and MRSA infections for appropriate antibiotic therapy.

Authors+Show Affiliations

Central Laboratory of the Second Affiliated Hospital, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, 550003, P.R. China. The Second Clinical College, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, 550003, P.R. China. Laboratory of Bacterial Infectious Disease of Experimental Center, Guizhou Provincial Centre for Disease Control and Prevention, 73 Bageyan Road, Guiyang, Guizhou, 550004, P.R. China.The Second Clinical College, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, 550003, P.R. China.Central Laboratory of the Second Affiliated Hospital, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, 550003, P.R. China.The Second Clinical College, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, 550003, P.R. China.Department of Clinical Laboratory Centre, The First People's Hospital of Guiyang, Guiyang, Guizhou, 55004, P.R. China.Laboratory of Bacterial Infectious Disease of Experimental Center, Guizhou Provincial Centre for Disease Control and Prevention, 73 Bageyan Road, Guiyang, Guizhou, 550004, P.R. China. zjumedjun@163.com.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32680560

Citation

Chen, Xu, et al. "The Rapid and Visual Detection of Methicillin-susceptible and Methicillin-resistant Staphylococcus Aureus Using Multiplex Loop-mediated Isothermal Amplification Linked to a Nanoparticle-based Lateral Flow Biosensor." Antimicrobial Resistance and Infection Control, vol. 9, no. 1, 2020, p. 111.
Chen X, Ma K, Yi X, et al. The rapid and visual detection of methicillin-susceptible and methicillin-resistant Staphylococcus aureus using multiplex loop-mediated isothermal amplification linked to a nanoparticle-based lateral flow biosensor. Antimicrob Resist Infect Control. 2020;9(1):111.
Chen, X., Ma, K., Yi, X., Xiong, L., Wang, Y., & Li, S. (2020). The rapid and visual detection of methicillin-susceptible and methicillin-resistant Staphylococcus aureus using multiplex loop-mediated isothermal amplification linked to a nanoparticle-based lateral flow biosensor. Antimicrobial Resistance and Infection Control, 9(1), 111. https://doi.org/10.1186/s13756-020-00774-x
Chen X, et al. The Rapid and Visual Detection of Methicillin-susceptible and Methicillin-resistant Staphylococcus Aureus Using Multiplex Loop-mediated Isothermal Amplification Linked to a Nanoparticle-based Lateral Flow Biosensor. Antimicrob Resist Infect Control. 2020 07 17;9(1):111. PubMed PMID: 32680560.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The rapid and visual detection of methicillin-susceptible and methicillin-resistant Staphylococcus aureus using multiplex loop-mediated isothermal amplification linked to a nanoparticle-based lateral flow biosensor. AU - Chen,Xu, AU - Ma,Kai, AU - Yi,Xu, AU - Xiong,Lijuan, AU - Wang,Yu, AU - Li,Shijun, Y1 - 2020/07/17/ PY - 2019/12/13/received PY - 2020/07/09/accepted PY - 2020/7/19/entrez PY - 2020/7/19/pubmed PY - 2021/7/13/medline KW - Lateral flow biosensor KW - Limit of detection KW - Loop-mediated isothermal amplification KW - MRSA KW - Staphylococcus aureus SP - 111 EP - 111 JF - Antimicrobial resistance and infection control JO - Antimicrob Resist Infect Control VL - 9 IS - 1 N2 - BACKGROUND: Staphylococcus aureus (S. aureus), including methicillin-susceptible S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA), is an eminent human pathogen that can colonize the human host and cause severe life-threatening infections. The development of a reliable, simple and rapid assay for detecting S. aureus and identifying MRSA is important for diagnosis and follow-up treatment. METHODS: A novel molecular diagnosis technique, named multiplex loop-mediated isothermal amplification linked to a nanoparticle-based lateral flow biosensor (m-LAMP-LFB), was applied to detect all S. aureus species and identify MRSA. Two sets of primers were designed based on the femA gene (S. aureus-specific gene) and the mecA gene (encoding penicillin-binding protein 2a), and the multiple-LAMP products were analyzed using LFB. The m-LAMP-LFB amplification conditions, including the target DNA concentration, reaction temperature and time, were optimized. The sensitivity and specificity of the m-LAMP-LFB method were tested in the current study, and the multiple-LAMP-LFB technology was applied to detect the MSSA and MRSA strains from clinical samples. RESULTS: The S. aureus- and MRSA-specific primers based on the femA and mecA genes allowed the multiple-LAMP technology to detect S. aureus and MRSA, respectively. The multiple-LAMP conditions were optimized at 63 °C for 40 min. The full process, including genomic DNA template preparation, LAMP, and product identification, could be achieved in 80 min. The limit of detection (LoD) of the multiple-LAMP assay for femA and mecA detection was 100 fg of genomic DNA template per reaction. The specificity of m-LAMP-LFB detection was 100 %, and no cross-reactions to non-S. aureus strains were observed. CONCLUSION: The multiple-LAMP-LFB technique developed in the current study is a reliable, simple, rapid, specific and sensitive method to identify MSSA and MRSA infections for appropriate antibiotic therapy. SN - 2047-2994 UR - https://www.unboundmedicine.com/medline/citation/32680560/The_rapid_and_visual_detection_of_methicillin_susceptible_and_methicillin_resistant_Staphylococcus_aureus_using_multiplex_loop_mediated_isothermal_amplification_linked_to_a_nanoparticle_based_lateral_flow_biosensor_ L2 - https://aricjournal.biomedcentral.com/articles/10.1186/s13756-020-00774-x DB - PRIME DP - Unbound Medicine ER -