Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS
Abstract Background Colistin (polymyxin E) and polymixin B are important bactericidal antibiotics used in the treatment of serious infections caused by multi-drug resistant Gram-negative organisms. Transferrable plasmid-mediated colistin resistance, conferred by the product of the mcr-1 gene, has em...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2019-02-01
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| Series: | Clinical Proteomics |
| Online Access: | http://link.springer.com/article/10.1186/s12014-019-9228-2 |
| _version_ | 1818307559514177536 |
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| author | Honghui Wang Yong Chen Jeffrey R. Strich Steven K. Drake Jung-Ho Youn Avi Z. Rosenberg Marjan Gucek Patrick T. McGann Anthony F. Suffredini John P. Dekker |
| author_facet | Honghui Wang Yong Chen Jeffrey R. Strich Steven K. Drake Jung-Ho Youn Avi Z. Rosenberg Marjan Gucek Patrick T. McGann Anthony F. Suffredini John P. Dekker |
| author_sort | Honghui Wang |
| collection | DOAJ |
| description | Abstract Background Colistin (polymyxin E) and polymixin B are important bactericidal antibiotics used in the treatment of serious infections caused by multi-drug resistant Gram-negative organisms. Transferrable plasmid-mediated colistin resistance, conferred by the product of the mcr-1 gene, has emerged as a global healthcare threat. Consequently, the rapid detection of the MCR-1 protein in clinical bacterial isolates has become increasingly important. We used a genoproteomic approach to identify unique peptides of the MCR-1 protein that could be detected rapidly by liquid chromatography tandem mass spectrometry (LC–MS/MS). Methods MCR-1 tryptic peptides that were efficiently ionized and readily detectable were characterized in a set of mcr-1-containing isolates with triple quadrupole LC–MS. Three optimal peptides were selected for the development of a rapid multiple reaction monitoring LC–MS/MS assay for the MCR-1 protein. To investigate the feasibility of rapid detection of the MCR-1 protein in bacterial isolates using this assay, a blinded 99-sample test set was built that included three additional mcr-1-containing clinical isolates tested in triplicate (9 samples) and 90 negative control isolates. Results All of the mcr-1-containing isolates in the test set were accurately identified with no false positive detections by three independent, blinded operators, yielding an overall performance of 100% sensitivity and specificity for multiple operators. Among the three peptides tested in this study, the best performing was DTFPQLAK. The isolate-to-result time for the assay as implemented is less than 90 min. Conclusions This work demonstrates the feasibility of rapid detection of the MCR-1 protein in bacterial isolates by LC–MS/MS. |
| first_indexed | 2024-12-13T07:00:18Z |
| format | Article |
| id | doaj.art-4331f9a41a214da3b0c2055a8a84357e |
| institution | Directory Open Access Journal |
| issn | 1542-6416 1559-0275 |
| language | English |
| last_indexed | 2024-12-13T07:00:18Z |
| publishDate | 2019-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Clinical Proteomics |
| spelling | doaj.art-4331f9a41a214da3b0c2055a8a84357e2022-12-21T23:55:56ZengBMCClinical Proteomics1542-64161559-02752019-02-0116111010.1186/s12014-019-9228-2Rapid detection of colistin resistance protein MCR-1 by LC–MS/MSHonghui Wang0Yong Chen1Jeffrey R. Strich2Steven K. Drake3Jung-Ho Youn4Avi Z. Rosenberg5Marjan Gucek6Patrick T. McGann7Anthony F. Suffredini8John P. Dekker9Critical Care Medicine Department, Clinical Center, National Institutes of HealthProteomics Core Facility, National Heart, Lung and Blood Institute, National Institutes of HealthCritical Care Medicine Department, Clinical Center, National Institutes of HealthCritical Care Medicine Department, Clinical Center, National Institutes of HealthDepartment of Laboratory Medicine, Clinical Center, Microbiology Service, National Institutes of HealthKidney Disease Section, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthProteomics Core Facility, National Heart, Lung and Blood Institute, National Institutes of HealthWalter Reed Army Institute of ResearchCritical Care Medicine Department, Clinical Center, National Institutes of HealthDepartment of Laboratory Medicine, Clinical Center, Microbiology Service, National Institutes of HealthAbstract Background Colistin (polymyxin E) and polymixin B are important bactericidal antibiotics used in the treatment of serious infections caused by multi-drug resistant Gram-negative organisms. Transferrable plasmid-mediated colistin resistance, conferred by the product of the mcr-1 gene, has emerged as a global healthcare threat. Consequently, the rapid detection of the MCR-1 protein in clinical bacterial isolates has become increasingly important. We used a genoproteomic approach to identify unique peptides of the MCR-1 protein that could be detected rapidly by liquid chromatography tandem mass spectrometry (LC–MS/MS). Methods MCR-1 tryptic peptides that were efficiently ionized and readily detectable were characterized in a set of mcr-1-containing isolates with triple quadrupole LC–MS. Three optimal peptides were selected for the development of a rapid multiple reaction monitoring LC–MS/MS assay for the MCR-1 protein. To investigate the feasibility of rapid detection of the MCR-1 protein in bacterial isolates using this assay, a blinded 99-sample test set was built that included three additional mcr-1-containing clinical isolates tested in triplicate (9 samples) and 90 negative control isolates. Results All of the mcr-1-containing isolates in the test set were accurately identified with no false positive detections by three independent, blinded operators, yielding an overall performance of 100% sensitivity and specificity for multiple operators. Among the three peptides tested in this study, the best performing was DTFPQLAK. The isolate-to-result time for the assay as implemented is less than 90 min. Conclusions This work demonstrates the feasibility of rapid detection of the MCR-1 protein in bacterial isolates by LC–MS/MS.http://link.springer.com/article/10.1186/s12014-019-9228-2 |
| spellingShingle | Honghui Wang Yong Chen Jeffrey R. Strich Steven K. Drake Jung-Ho Youn Avi Z. Rosenberg Marjan Gucek Patrick T. McGann Anthony F. Suffredini John P. Dekker Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS Clinical Proteomics |
| title | Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
| title_full | Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
| title_fullStr | Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
| title_full_unstemmed | Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
| title_short | Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
| title_sort | rapid detection of colistin resistance protein mcr 1 by lc ms ms |
| url | http://link.springer.com/article/10.1186/s12014-019-9228-2 |
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