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Development of real-time PCR assay for detection of total bacteria in beverage emulsions
The rapid growth and expansion of the soft drinks market and the necessity to meet and maintain the consumers’ expectations of having high quality products safe for consumption, have both drawn the attention to the need for rapid and sensitive methods for the detection of potential microbial contami...
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| Format: | Thesis |
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2015
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| _version_ | 1867613407645007872 |
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| access_status_str | Open Access |
| author | Elsisi, Essam Gamaleldin |
| author_browse | Elsisi, Essam Gamaleldin |
| author_facet | Elsisi, Essam Gamaleldin |
| author_sort | Elsisi, Essam Gamaleldin |
| collection | Thesis |
| dc_rights_str_mv | The author retains all rights with regard to copyright. The author certifies that written permission from the owner(s) of third-party copyrighted matter included in the thesis, dissertation, paper, or record of study has been obtained. The author further certifies that IRB approval has been obtained for this thesis, or that IRB approval is not necessary for this thesis. Insofar as this thesis, dissertation, paper, or record of study is an educational record as defined in the Family Educational Rights and Privacy Act (FERPA) (20 USC 1232g), the author has granted consent to disclosure of it to anyone who requests a copy. |
| description | The rapid growth and expansion of the soft drinks market and the necessity to meet and maintain the consumers’ expectations of having high quality products safe for consumption, have both drawn the attention to the need for rapid and sensitive methods for the detection of potential microbial contaminations. This has made the current conventional culture-based methods inconvenient due to the relatively long periods of time they need to yield results, in addition to their relatively low sensitivity. In contrast, real-time PCR is a rapid and sensitive molecular detection technique, capable of providing quick detection and quantification methods of specific DNA sequences even if the quantity of the starting material is small. In this study, a real-time PCR assay for the determination of total bacteria in one of the microbiologically sensitive constituents of soft drinks, called beverage emulsions, was successfully developed. This included the development of a DNA extraction protocol and the selection of a set of universal primers targeting a conserved region in the 16S rDNA of bacteria. The quantification strategy was based on a standard curve and a calculation method for the conversion of the determined DNA concentrations to bacterial cells numbers. This enabled the sensitive determination of total bacteria in beverage emulsions in the range between 10 fg/µL and 100 ng/µL, corresponding to 2 and 2 x 107 cells of Escherichia coli, respectively, in 6 to 8 hours instead of 7 days required by the pour plate method. Further optimization of the developed assay may allow the determination of viable bacterial cells, which will extend the scope of the developed assay applications in this industry, in the future. |
| format | Thesis |
| id | oai:fount.aucegypt.edu:etds-1100 |
| institution | American University in Cairo (Egypt) |
| last_indexed | 2026-06-10T12:35:39.635Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from AUC Knowledge Fountain — bepress |
| publishDate | 2015 |
| publishDateRange | 2015 |
| publishDateSort | 2015 |
| publisher | AUC Knowledge Fountain |
| publisherStr | AUC Knowledge Fountain |
| record_format | dspace |
| source_str | AUC Knowledge Fountain — bepress |
| spelling | oai:fount.aucegypt.edu:etds-1100 Development of real-time PCR assay for detection of total bacteria in beverage emulsions Elsisi, Essam Gamaleldin The rapid growth and expansion of the soft drinks market and the necessity to meet and maintain the consumers’ expectations of having high quality products safe for consumption, have both drawn the attention to the need for rapid and sensitive methods for the detection of potential microbial contaminations. This has made the current conventional culture-based methods inconvenient due to the relatively long periods of time they need to yield results, in addition to their relatively low sensitivity. In contrast, real-time PCR is a rapid and sensitive molecular detection technique, capable of providing quick detection and quantification methods of specific DNA sequences even if the quantity of the starting material is small. In this study, a real-time PCR assay for the determination of total bacteria in one of the microbiologically sensitive constituents of soft drinks, called beverage emulsions, was successfully developed. This included the development of a DNA extraction protocol and the selection of a set of universal primers targeting a conserved region in the 16S rDNA of bacteria. The quantification strategy was based on a standard curve and a calculation method for the conversion of the determined DNA concentrations to bacterial cells numbers. This enabled the sensitive determination of total bacteria in beverage emulsions in the range between 10 fg/µL and 100 ng/µL, corresponding to 2 and 2 x 107 cells of Escherichia coli, respectively, in 6 to 8 hours instead of 7 days required by the pour plate method. Further optimization of the developed assay may allow the determination of viable bacterial cells, which will extend the scope of the developed assay applications in this industry, in the future. 2015-02-01T08:00:00Z thesis application/pdf https://fount.aucegypt.edu/etds/101 https://fount.aucegypt.edu/context/etds/article/1100/viewcontent/M.Sc._20Thesis_20__20Essam_20Elsisi.pdf The author retains all rights with regard to copyright. The author certifies that written permission from the owner(s) of third-party copyrighted matter included in the thesis, dissertation, paper, or record of study has been obtained. The author further certifies that IRB approval has been obtained for this thesis, or that IRB approval is not necessary for this thesis. Insofar as this thesis, dissertation, paper, or record of study is an educational record as defined in the Family Educational Rights and Privacy Act (FERPA) (20 USC 1232g), the author has granted consent to disclosure of it to anyone who requests a copy. Theses and Dissertations AUC Knowledge Fountain Real-Time PCR Total Bacteria |
| spellingShingle | Real-Time PCR Total Bacteria Elsisi, Essam Gamaleldin Development of real-time PCR assay for detection of total bacteria in beverage emulsions |
| title | Development of real-time PCR assay for detection of total bacteria in beverage emulsions |
| title_full | Development of real-time PCR assay for detection of total bacteria in beverage emulsions |
| title_fullStr | Development of real-time PCR assay for detection of total bacteria in beverage emulsions |
| title_full_unstemmed | Development of real-time PCR assay for detection of total bacteria in beverage emulsions |
| title_short | Development of real-time PCR assay for detection of total bacteria in beverage emulsions |
| title_sort | development of real time pcr assay for detection of total bacteria in beverage emulsions |
| topic | Real-Time PCR Total Bacteria |
| url | https://fount.aucegypt.edu/etds/101 https://fount.aucegypt.edu/context/etds/article/1100/viewcontent/M.Sc._20Thesis_20__20Essam_20Elsisi.pdf |
| work_keys_str_mv | AT elsisiessamgamaleldin developmentofrealtimepcrassayfordetectionoftotalbacteriainbeverageemulsions |