Sample preparation using multiple microbial pattern recognition proteins and magnetic bead ratcheting
Sample preparation (i.e., isolation and purification of pathogens from samples) remains one of the bottlenecks limiting deployment of diagnostic tests. Two recent examples highlighting these limitations include the difficulties in widespread nucleic acid testing during COVID-19 and in identifying dr...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-08-01
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| Series: | SLAS Technology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2472630325000731 |
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| author | Jessica Martinez Dong Jin M. Park Samantha Abate James Hill George Downey Craig Galligan Tyler Hammond Michael T. McCurdy Surekha Gurung Shanjana Shawon Ralf Lenigk Kuangwen Hsieh Michael Super Erik Kvam Tza-Huei Wang Coleman Murray Chris Puleo |
| author_facet | Jessica Martinez Dong Jin M. Park Samantha Abate James Hill George Downey Craig Galligan Tyler Hammond Michael T. McCurdy Surekha Gurung Shanjana Shawon Ralf Lenigk Kuangwen Hsieh Michael Super Erik Kvam Tza-Huei Wang Coleman Murray Chris Puleo |
| author_sort | Jessica Martinez |
| collection | DOAJ |
| description | Sample preparation (i.e., isolation and purification of pathogens from samples) remains one of the bottlenecks limiting deployment of diagnostic tests. Two recent examples highlighting these limitations include the difficulties in widespread nucleic acid testing during COVID-19 and in identifying drug-resistant infections. Unmet needs include systems that work with different bodily fluids and samples, multiple types of suspected pathogen, and function in a rapid and semi-automated fashion. Advances such as these could accelerate the deployment of novel diagnostic tests by eliminating upstream sample preparation bottlenecks.Herein, we tested the feasibility of combining multiple pathogen-binding paramagnetic beads with magnetic ratcheting-based enrichment to directly isolate microbes from samples. We demonstrate effective use of three different paramagnetic bead-conjugated proteins (mannose binding lection [MBL], C-reactive protein [CRP], and dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin [DC-SIGN]) to capture 18 different bacteria species and model virus particles/peptides. We used magnetic ratcheting to isolate bead-bound microbes from milliliters (mLs) of sample at concentrations of 4 – 4000 microbes per mL. The flow-through system was operated up to 1 mL per minute and enabled pathogen isolation from 10 mL samples in <30 min. Demonstration of post-ratcheting PCR-based microbe analysis was also performed, showing that the technology may have applicability across different infectious agents, sample types/volumes, and analytical assays. Based on these results, further studies are warranted to test clinical samples, compare results to current gold-standard diagnostic methods, and test the sample preparation technologies across additional pathogen types. |
| format | Article |
| id | doaj-art-03a1f18836474f409bf4c5fd5916d945 |
| institution | Kabale University |
| issn | 2472-6303 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | SLAS Technology |
| spelling | doaj-art-03a1f18836474f409bf4c5fd5916d9452025-08-20T04:00:27ZengElsevierSLAS Technology2472-63032025-08-013310031510.1016/j.slast.2025.100315Sample preparation using multiple microbial pattern recognition proteins and magnetic bead ratchetingJessica Martinez0Dong Jin M. Park1Samantha Abate2James Hill3George Downey4Craig Galligan5Tyler Hammond6Michael T. McCurdy7Surekha Gurung8Shanjana Shawon9Ralf Lenigk10Kuangwen Hsieh11Michael Super12Erik Kvam13Tza-Huei Wang14Coleman Murray15Chris Puleo16General Electric Healthcare Technology and Innovation Center, 1 Research Circle, Niskayuna, NY 12309, USAJohns Hopkins University, Baltimore, MD 21218, USAGeneral Electric Healthcare Technology and Innovation Center, 1 Research Circle, Niskayuna, NY 12309, USABOA Biomedical, 20 University Rd., Cambridge, MA 02138, USABOA Biomedical, 20 University Rd., Cambridge, MA 02138, USAGeneral Electric Healthcare Technology and Innovation Center, 1 Research Circle, Niskayuna, NY 12309, USAGeneral Electric Healthcare Technology and Innovation Center, 1 Research Circle, Niskayuna, NY 12309, USABOA Biomedical, 20 University Rd., Cambridge, MA 02138, USA; University of Maryland School of Medicine, Baltimore, MD 21201, USARensselaer Polytechnic Institute, Troy, NY 12180, USARensselaer Polytechnic Institute, Troy, NY 12180, USAGeneral Electric Healthcare Technology and Innovation Center, 1 Research Circle, Niskayuna, NY 12309, USAJohns Hopkins University, Baltimore, MD 21218, USAWyss Institute, Harvard University, 201 Brookline Ave, Boston, MA 02215, USAGeneral Electric Healthcare Technology and Innovation Center, 1 Research Circle, Niskayuna, NY 12309, USAJohns Hopkins University, Baltimore, MD 21218, USAFerrologix, 23328 Barfield Dr, Valencia, CA 91354, USARensselaer Polytechnic Institute, Troy, NY 12180, USA; Corresponding author.Sample preparation (i.e., isolation and purification of pathogens from samples) remains one of the bottlenecks limiting deployment of diagnostic tests. Two recent examples highlighting these limitations include the difficulties in widespread nucleic acid testing during COVID-19 and in identifying drug-resistant infections. Unmet needs include systems that work with different bodily fluids and samples, multiple types of suspected pathogen, and function in a rapid and semi-automated fashion. Advances such as these could accelerate the deployment of novel diagnostic tests by eliminating upstream sample preparation bottlenecks.Herein, we tested the feasibility of combining multiple pathogen-binding paramagnetic beads with magnetic ratcheting-based enrichment to directly isolate microbes from samples. We demonstrate effective use of three different paramagnetic bead-conjugated proteins (mannose binding lection [MBL], C-reactive protein [CRP], and dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin [DC-SIGN]) to capture 18 different bacteria species and model virus particles/peptides. We used magnetic ratcheting to isolate bead-bound microbes from milliliters (mLs) of sample at concentrations of 4 – 4000 microbes per mL. The flow-through system was operated up to 1 mL per minute and enabled pathogen isolation from 10 mL samples in <30 min. Demonstration of post-ratcheting PCR-based microbe analysis was also performed, showing that the technology may have applicability across different infectious agents, sample types/volumes, and analytical assays. Based on these results, further studies are warranted to test clinical samples, compare results to current gold-standard diagnostic methods, and test the sample preparation technologies across additional pathogen types.http://www.sciencedirect.com/science/article/pii/S2472630325000731Magnetic ratchetingDiagnosticsPathogenInfectious diseaseSample preparationAutomation |
| spellingShingle | Jessica Martinez Dong Jin M. Park Samantha Abate James Hill George Downey Craig Galligan Tyler Hammond Michael T. McCurdy Surekha Gurung Shanjana Shawon Ralf Lenigk Kuangwen Hsieh Michael Super Erik Kvam Tza-Huei Wang Coleman Murray Chris Puleo Sample preparation using multiple microbial pattern recognition proteins and magnetic bead ratcheting SLAS Technology Magnetic ratcheting Diagnostics Pathogen Infectious disease Sample preparation Automation |
| title | Sample preparation using multiple microbial pattern recognition proteins and magnetic bead ratcheting |
| title_full | Sample preparation using multiple microbial pattern recognition proteins and magnetic bead ratcheting |
| title_fullStr | Sample preparation using multiple microbial pattern recognition proteins and magnetic bead ratcheting |
| title_full_unstemmed | Sample preparation using multiple microbial pattern recognition proteins and magnetic bead ratcheting |
| title_short | Sample preparation using multiple microbial pattern recognition proteins and magnetic bead ratcheting |
| title_sort | sample preparation using multiple microbial pattern recognition proteins and magnetic bead ratcheting |
| topic | Magnetic ratcheting Diagnostics Pathogen Infectious disease Sample preparation Automation |
| url | http://www.sciencedirect.com/science/article/pii/S2472630325000731 |
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