A magnetic epitope-imprinted microsphere used for selective separation and rapid detection of SHV-type β-lactamases in bacteria: a novel strategy of antimicrobial resistance detection
Abstract Background The production of β-lactamases is the most prevalent resistance mechanism for β-lactam antibiotics in Gram-negative bacteria. Presently, over 4900 β-lactamases have been discovered, and they are categorized into hundreds of families. In each enzyme family, amino acid substitution...
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2024-11-01
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| Series: | Journal of Nanobiotechnology |
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| Online Access: | https://doi.org/10.1186/s12951-024-02949-9 |
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| author | Yusun Zhou Kunqi Wang Lele Li Hui Li Qingwu Tian Baosheng Ge Yuanyuan Chi Xiaotong Xu Shuhui Liu Meng Han Tingting Zhou Yuanqi Zhu Qing Wang Bing Yu |
| author_facet | Yusun Zhou Kunqi Wang Lele Li Hui Li Qingwu Tian Baosheng Ge Yuanyuan Chi Xiaotong Xu Shuhui Liu Meng Han Tingting Zhou Yuanqi Zhu Qing Wang Bing Yu |
| author_sort | Yusun Zhou |
| collection | DOAJ |
| description | Abstract Background The production of β-lactamases is the most prevalent resistance mechanism for β-lactam antibiotics in Gram-negative bacteria. Presently, over 4900 β-lactamases have been discovered, and they are categorized into hundreds of families. In each enzyme family, amino acid substitutions result in subtle changes to enzyme hydrolysis profiles; in contrast, certain conserved sequences retained by all of the family members can serve as important markers for enzyme family identification. Results The SHV family was chosen as the study object. First, a unique 10-mer peptide was identified as SHV family's epitope by an approach of protein fingerprint analysis. Then, an SHV-specific magnetic epitope-imprinted gel polymer (MEI-GP) was prepared by an epitope surface imprinting technique, and its sorption behavior and recognition mechanism for template epitope and SHV were both elaborated. Finally, the MEI-GP was successfully applied to selectively extract SHV from bacteria, and the extracted SHV was submitted to MALDI-TOF MS for specific determination. By following this strategy, other β-lactamase families can also be specifically detected. According to the molecular weight displayed in mass spectra, the kind of β-lactamase and its associated hydrolysis profile on β-lactams can be easily identified. Based on this, an initial drug option scheme can be quickly formulated for antimicrobial therapy. From protein extraction to medication guidance reporting, the mean time to detection (MTTD) was less than 2 h, which is much faster than conventional phenotype-based methods (at least 16–20 h) and gene-based techniques (usually about 8 h). Conclusions This enzyme-specific detection strategy combined the specificity of epitope imprinting with the sensitivity of mass spectrometry, enabling β-lactamase to be selectively extracted from bacteria and clearly presented in mass spectra. Compared with other drug resistance detection methods, this technique has good specificity, high sensitivity (≤ 15 mg of bacteria), a short MTTD (less than 2 h), and simple operation, and therefore has a broad application prospect in clinical medicine. Graphical Abstract |
| format | Article |
| id | doaj-art-981e31ee914a447e994defca7f8d88df |
| institution | Kabale University |
| issn | 1477-3155 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | BMC |
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| series | Journal of Nanobiotechnology |
| spelling | doaj-art-981e31ee914a447e994defca7f8d88df2024-11-10T12:41:23ZengBMCJournal of Nanobiotechnology1477-31552024-11-0122111910.1186/s12951-024-02949-9A magnetic epitope-imprinted microsphere used for selective separation and rapid detection of SHV-type β-lactamases in bacteria: a novel strategy of antimicrobial resistance detectionYusun Zhou0Kunqi Wang1Lele Li2Hui Li3Qingwu Tian4Baosheng Ge5Yuanyuan Chi6Xiaotong Xu7Shuhui Liu8Meng Han9Tingting Zhou10Yuanqi Zhu11Qing Wang12Bing Yu13Department of Clinical Laboratory, The Affiliated Hospital of Qingdao UniversityDepartment of Clinical Laboratory, The Affiliated Hospital of Qingdao UniversityDepartment of Clinical Laboratory, The Affiliated Hospital of Qingdao UniversityDepartment of Clinical Laboratory, The Affiliated Hospital of Qingdao UniversityDepartment of Clinical Laboratory, The Affiliated Hospital of Qingdao UniversityState Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China)Qingdao Women and Children’s HospitalDepartment of Pediatric Emergency, The Affiliated Hospital of Qingdao UniversityDepartment of Clinical Laboratory, The Affiliated Hospital of Qingdao UniversityQingdao Women and Children’s HospitalDepartment of Clinical Laboratory, The Affiliated Hospital of Qingdao UniversityDepartment of Clinical Laboratory, The Affiliated Hospital of Qingdao UniversityDepartment of Clinical Laboratory, The Affiliated Hospital of Qingdao UniversityCollege of Chemistry and Chemical Engineering, Institute of Biomedical Materials and Engineering, College of Materials Science and Engineering, Qingdao UniversityAbstract Background The production of β-lactamases is the most prevalent resistance mechanism for β-lactam antibiotics in Gram-negative bacteria. Presently, over 4900 β-lactamases have been discovered, and they are categorized into hundreds of families. In each enzyme family, amino acid substitutions result in subtle changes to enzyme hydrolysis profiles; in contrast, certain conserved sequences retained by all of the family members can serve as important markers for enzyme family identification. Results The SHV family was chosen as the study object. First, a unique 10-mer peptide was identified as SHV family's epitope by an approach of protein fingerprint analysis. Then, an SHV-specific magnetic epitope-imprinted gel polymer (MEI-GP) was prepared by an epitope surface imprinting technique, and its sorption behavior and recognition mechanism for template epitope and SHV were both elaborated. Finally, the MEI-GP was successfully applied to selectively extract SHV from bacteria, and the extracted SHV was submitted to MALDI-TOF MS for specific determination. By following this strategy, other β-lactamase families can also be specifically detected. According to the molecular weight displayed in mass spectra, the kind of β-lactamase and its associated hydrolysis profile on β-lactams can be easily identified. Based on this, an initial drug option scheme can be quickly formulated for antimicrobial therapy. From protein extraction to medication guidance reporting, the mean time to detection (MTTD) was less than 2 h, which is much faster than conventional phenotype-based methods (at least 16–20 h) and gene-based techniques (usually about 8 h). Conclusions This enzyme-specific detection strategy combined the specificity of epitope imprinting with the sensitivity of mass spectrometry, enabling β-lactamase to be selectively extracted from bacteria and clearly presented in mass spectra. Compared with other drug resistance detection methods, this technique has good specificity, high sensitivity (≤ 15 mg of bacteria), a short MTTD (less than 2 h), and simple operation, and therefore has a broad application prospect in clinical medicine. Graphical Abstracthttps://doi.org/10.1186/s12951-024-02949-9Antimicrobial resistance detectionβ-LactamasesSHV familyEpitope imprintingProtein-specific recognition |
| spellingShingle | Yusun Zhou Kunqi Wang Lele Li Hui Li Qingwu Tian Baosheng Ge Yuanyuan Chi Xiaotong Xu Shuhui Liu Meng Han Tingting Zhou Yuanqi Zhu Qing Wang Bing Yu A magnetic epitope-imprinted microsphere used for selective separation and rapid detection of SHV-type β-lactamases in bacteria: a novel strategy of antimicrobial resistance detection Journal of Nanobiotechnology Antimicrobial resistance detection β-Lactamases SHV family Epitope imprinting Protein-specific recognition |
| title | A magnetic epitope-imprinted microsphere used for selective separation and rapid detection of SHV-type β-lactamases in bacteria: a novel strategy of antimicrobial resistance detection |
| title_full | A magnetic epitope-imprinted microsphere used for selective separation and rapid detection of SHV-type β-lactamases in bacteria: a novel strategy of antimicrobial resistance detection |
| title_fullStr | A magnetic epitope-imprinted microsphere used for selective separation and rapid detection of SHV-type β-lactamases in bacteria: a novel strategy of antimicrobial resistance detection |
| title_full_unstemmed | A magnetic epitope-imprinted microsphere used for selective separation and rapid detection of SHV-type β-lactamases in bacteria: a novel strategy of antimicrobial resistance detection |
| title_short | A magnetic epitope-imprinted microsphere used for selective separation and rapid detection of SHV-type β-lactamases in bacteria: a novel strategy of antimicrobial resistance detection |
| title_sort | magnetic epitope imprinted microsphere used for selective separation and rapid detection of shv type β lactamases in bacteria a novel strategy of antimicrobial resistance detection |
| topic | Antimicrobial resistance detection β-Lactamases SHV family Epitope imprinting Protein-specific recognition |
| url | https://doi.org/10.1186/s12951-024-02949-9 |
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