Simple dual filter workflow for facilitating blood culture-free and sensitive detection of pathogenic bacteria from blood
Abstract Bloodstream infections (BSI) caused by pathogenic bacteria are a global health threat, highlighting the need for improved diagnostic methods. Current methods, such as mass spectrometry, depend on time-consuming blood cultures, which hinder timely diagnosis. Nucleic acid amplification tests...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-07-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-025-08987-z |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849237726892654592 |
|---|---|
| author | Dong Jin M. Park Tianqi Wu Liben Chen Pei-Wei Lee Kuangwen Hsieh Tza-Huei Wang |
| author_facet | Dong Jin M. Park Tianqi Wu Liben Chen Pei-Wei Lee Kuangwen Hsieh Tza-Huei Wang |
| author_sort | Dong Jin M. Park |
| collection | DOAJ |
| description | Abstract Bloodstream infections (BSI) caused by pathogenic bacteria are a global health threat, highlighting the need for improved diagnostic methods. Current methods, such as mass spectrometry, depend on time-consuming blood cultures, which hinder timely diagnosis. Nucleic acid amplification tests (NAATs) offer potential for sensitive bacteria detection but are bottlenecked by the inability to isolate bacteria at low concentrations from blood. This study introduces a facile dual filter workflow for isolating low concentrations of bacteria from blood without blood culture. The workflow, which involves blood dilution and osmolysis, blood cell filtration, chemical and enzymatic lysis, and filter-based bacteria capture, can be completed in ~ 30 min without special equipment. It readily combines with downstream broad-based bacteria detection via PCR and high-resolution melt (HRM), as well as pheno-molecular antimicrobial susceptibility testing (AST) via PCR. When coupled with species-specific PCR, ~ 10 CFU of bacteria in 0.5 mL of blood could be detected. The small sample volume is particularly attractive because it is comparable to those obtained from fingerstick collections or pediatric patients. With further optimization, testing with a wider range of bacteria, and integration with rapid point-of-care testing, this simple dual filter workflow can be a key driver for advancing blood culture-free bacterial BSI diagnosis, especially in resource-limited settings. |
| format | Article |
| id | doaj-art-e540fad6403941f185904a8bb1d24b8f |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-e540fad6403941f185904a8bb1d24b8f2025-08-20T04:01:52ZengNature PortfolioScientific Reports2045-23222025-07-0115111110.1038/s41598-025-08987-zSimple dual filter workflow for facilitating blood culture-free and sensitive detection of pathogenic bacteria from bloodDong Jin M. Park0Tianqi Wu1Liben Chen2Pei-Wei Lee3Kuangwen Hsieh4Tza-Huei Wang5Department of Biomedical Engineering, Johns Hopkins UniversityDepartment of Biomedical Engineering, Johns Hopkins UniversityDepartment of Mechanical Engineering, Johns Hopkins UniversityDepartment of Mechanical Engineering, Johns Hopkins UniversityDepartment of Mechanical Engineering, Johns Hopkins UniversityDepartment of Biomedical Engineering, Johns Hopkins UniversityAbstract Bloodstream infections (BSI) caused by pathogenic bacteria are a global health threat, highlighting the need for improved diagnostic methods. Current methods, such as mass spectrometry, depend on time-consuming blood cultures, which hinder timely diagnosis. Nucleic acid amplification tests (NAATs) offer potential for sensitive bacteria detection but are bottlenecked by the inability to isolate bacteria at low concentrations from blood. This study introduces a facile dual filter workflow for isolating low concentrations of bacteria from blood without blood culture. The workflow, which involves blood dilution and osmolysis, blood cell filtration, chemical and enzymatic lysis, and filter-based bacteria capture, can be completed in ~ 30 min without special equipment. It readily combines with downstream broad-based bacteria detection via PCR and high-resolution melt (HRM), as well as pheno-molecular antimicrobial susceptibility testing (AST) via PCR. When coupled with species-specific PCR, ~ 10 CFU of bacteria in 0.5 mL of blood could be detected. The small sample volume is particularly attractive because it is comparable to those obtained from fingerstick collections or pediatric patients. With further optimization, testing with a wider range of bacteria, and integration with rapid point-of-care testing, this simple dual filter workflow can be a key driver for advancing blood culture-free bacterial BSI diagnosis, especially in resource-limited settings.https://doi.org/10.1038/s41598-025-08987-zBloodstream infectionsBlood processingBlood culture-freeSensitivityRapid diagnosisResource-limited settings |
| spellingShingle | Dong Jin M. Park Tianqi Wu Liben Chen Pei-Wei Lee Kuangwen Hsieh Tza-Huei Wang Simple dual filter workflow for facilitating blood culture-free and sensitive detection of pathogenic bacteria from blood Scientific Reports Bloodstream infections Blood processing Blood culture-free Sensitivity Rapid diagnosis Resource-limited settings |
| title | Simple dual filter workflow for facilitating blood culture-free and sensitive detection of pathogenic bacteria from blood |
| title_full | Simple dual filter workflow for facilitating blood culture-free and sensitive detection of pathogenic bacteria from blood |
| title_fullStr | Simple dual filter workflow for facilitating blood culture-free and sensitive detection of pathogenic bacteria from blood |
| title_full_unstemmed | Simple dual filter workflow for facilitating blood culture-free and sensitive detection of pathogenic bacteria from blood |
| title_short | Simple dual filter workflow for facilitating blood culture-free and sensitive detection of pathogenic bacteria from blood |
| title_sort | simple dual filter workflow for facilitating blood culture free and sensitive detection of pathogenic bacteria from blood |
| topic | Bloodstream infections Blood processing Blood culture-free Sensitivity Rapid diagnosis Resource-limited settings |
| url | https://doi.org/10.1038/s41598-025-08987-z |
| work_keys_str_mv | AT dongjinmpark simpledualfilterworkflowforfacilitatingbloodculturefreeandsensitivedetectionofpathogenicbacteriafromblood AT tianqiwu simpledualfilterworkflowforfacilitatingbloodculturefreeandsensitivedetectionofpathogenicbacteriafromblood AT libenchen simpledualfilterworkflowforfacilitatingbloodculturefreeandsensitivedetectionofpathogenicbacteriafromblood AT peiweilee simpledualfilterworkflowforfacilitatingbloodculturefreeandsensitivedetectionofpathogenicbacteriafromblood AT kuangwenhsieh simpledualfilterworkflowforfacilitatingbloodculturefreeandsensitivedetectionofpathogenicbacteriafromblood AT tzahueiwang simpledualfilterworkflowforfacilitatingbloodculturefreeandsensitivedetectionofpathogenicbacteriafromblood |