Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce
A sensitive, high-throughput, and cost-effective method for screening bacterial pathogens in the environment was developed. A variety of environmental samples, including aerosols, soil of various types (sand, sand/clay mix, and clay), wastewater, and vegetable surface (modeled by tomato), were conco...
| Main Authors: | , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
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Springer International Publishing
2017
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| Online Access: | http://hdl.handle.net/1721.1/107155 |
| _version_ | 1811069957610405888 |
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| author | Orlofsky, Ezra Benami, Maya Gross, Amit Gillor, Osnat Dutt, Michelle M. |
| author2 | Massachusetts Institute of Technology. Department of Civil and Environmental Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Civil and Environmental Engineering Orlofsky, Ezra Benami, Maya Gross, Amit Gillor, Osnat Dutt, Michelle M. |
| author_sort | Orlofsky, Ezra |
| collection | MIT |
| description | A sensitive, high-throughput, and cost-effective method for screening bacterial pathogens in the environment was developed. A variety of environmental samples, including aerosols, soil of various types (sand, sand/clay mix, and clay), wastewater, and vegetable surface (modeled by tomato), were concomitantly spiked with Salmonella enterica and/or Pseudomonas aeruginosa to determine recovery rates and limits of detection. The various matrices were first enriched with a general pre-enrichment broth in a dilution series and then enumerated by most probable number (MPN) estimation using quantitative PCR for rapid screening of amplicon presence. Soil and aerosols were then tested in non-spiked environmental samples, as these matrices are prone to large experimental variation. Limit of detection in the various soil types was 1–3 colony-forming units (CFU) g[superscript −1]; on vegetable surface, 5 CFU per tomato; in treated wastewater, 5 CFU L[superscript −1]; and in aerosols, >300 CFU mL[superscript −1]. Our method accurately identified S. enterica in non-spiked environmental soil samples within a day, while traditional methods took 4 to 5 days and required sorting through biochemically and morphologically similar species. Likewise, our method successfully identified P. aeruginosa in non-spiked aerosols generated by a domestic wastewater treatment system. The obtained results suggest that the developed method presents a broad approach for the rapid, efficient, and reliable detection of relatively low densities of pathogenic organisms in challenging environmental samples. |
| first_indexed | 2024-09-23T08:19:34Z |
| format | Article |
| id | mit-1721.1/107155 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T08:19:34Z |
| publishDate | 2017 |
| publisher | Springer International Publishing |
| record_format | dspace |
| spelling | mit-1721.1/1071552022-09-30T09:03:24Z Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce Orlofsky, Ezra Benami, Maya Gross, Amit Gillor, Osnat Dutt, Michelle M. Massachusetts Institute of Technology. Department of Civil and Environmental Engineering Dutt, Michelle M. A sensitive, high-throughput, and cost-effective method for screening bacterial pathogens in the environment was developed. A variety of environmental samples, including aerosols, soil of various types (sand, sand/clay mix, and clay), wastewater, and vegetable surface (modeled by tomato), were concomitantly spiked with Salmonella enterica and/or Pseudomonas aeruginosa to determine recovery rates and limits of detection. The various matrices were first enriched with a general pre-enrichment broth in a dilution series and then enumerated by most probable number (MPN) estimation using quantitative PCR for rapid screening of amplicon presence. Soil and aerosols were then tested in non-spiked environmental samples, as these matrices are prone to large experimental variation. Limit of detection in the various soil types was 1–3 colony-forming units (CFU) g[superscript −1]; on vegetable surface, 5 CFU per tomato; in treated wastewater, 5 CFU L[superscript −1]; and in aerosols, >300 CFU mL[superscript −1]. Our method accurately identified S. enterica in non-spiked environmental soil samples within a day, while traditional methods took 4 to 5 days and required sorting through biochemically and morphologically similar species. Likewise, our method successfully identified P. aeruginosa in non-spiked aerosols generated by a domestic wastewater treatment system. The obtained results suggest that the developed method presents a broad approach for the rapid, efficient, and reliable detection of relatively low densities of pathogenic organisms in challenging environmental samples. United States-Israel Binational Agricultural Research and Development Fund (Grant No. CP-9033-09) MIT International Science and Technology Initiatives Kraft Foods Company 2017-02-24T21:10:12Z 2017-02-24T21:10:12Z 2015-08 2015-05 2016-08-18T15:40:33Z Article http://purl.org/eprint/type/JournalArticle 0049-6979 1573-2932 http://hdl.handle.net/1721.1/107155 Orlofsky, Ezra et al. “Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce.” Water, Air, & Soil Pollution 226.9 (2015): n. pag. en http://dx.doi.org/10.1007/s11270-015-2560-x Water, Air, & Soil Pollution Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. Springer International Publishing Switzerland application/pdf Springer International Publishing Springer International Publishing |
| spellingShingle | Orlofsky, Ezra Benami, Maya Gross, Amit Gillor, Osnat Dutt, Michelle M. Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce |
| title | Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce |
| title_full | Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce |
| title_fullStr | Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce |
| title_full_unstemmed | Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce |
| title_short | Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce |
| title_sort | rapid mpn qpcr screening for pathogens in air soil water and agricultural produce |
| url | http://hdl.handle.net/1721.1/107155 |
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