A novel approach for simultaneous detection of the most common food-borne pathogens by multiplex qPCR
Food contaminated with bacterial pathogens is a great threat to human health and food spoilage, having an impact on public health and the food industry. Research in food safety seeks to develop a practical, rapid, and sensitive detection technique for food-borne pathogens. In the past few decades,...
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| Format: | Article |
| Language: | English |
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Association of Basic Medical Sciences of Federation of Bosnia and Herzegovina
2023-07-01
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| Series: | Biomolecules & Biomedicine |
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| Online Access: | https://www.bjbms.org/ojs/index.php/bjbms/article/view/8693 |
| _version_ | 1797261382864863232 |
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| author | Emir Hodzic Aida Glavinic Cara Wademan |
| author_facet | Emir Hodzic Aida Glavinic Cara Wademan |
| author_sort | Emir Hodzic |
| collection | DOAJ |
| description |
Food contaminated with bacterial pathogens is a great threat to human health and food spoilage, having an impact on public health and the food industry. Research in food safety seeks to develop a practical, rapid, and sensitive detection technique for food-borne pathogens. In the past few decades, real-time quantitative polymerase chain reaction (qPCR) has been developed, and multiplex qPCR is a preferred feature. Multiplex qPCR enables the simultaneous amplification of many targets of interest in one reaction by using more than one pair of primers. In this study, we have developed and evaluated a hydrolysis (TaqMan) probe-based system for simultaneous detection of eight of the most common food-borne pathogens in a single-step procedure by multiplex qPCR. A multicolor combinational probe coding (MCPC) strategy was utilized that allows multiple fluorophores to label different probes in combinatorial manner. This strategy enabled simultaneous detection, identification, and quantification of targeted genes. The efficiency of the individual qPCR reactions for each target gene had values comparable to those established for multiplex qPCR, with detection limits of approximately < 10 copies of DNA per reaction. Pathogen load helps to predict bacteriological quality status in food products and serves to validate the efficiency of procedures to minimize or eliminate their presence, so newly developed multiplex qPCR was quantitative for each pathogen. During sample preparation, a step to concentrate the target organism from a relatively large sample size, remove all potential PCR inhibitors, and yield samples in a volume suitable for qPCR was incorporated.
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| first_indexed | 2024-04-24T23:40:20Z |
| format | Article |
| id | doaj.art-9f3f3aeef5ff466b8b4a2894223767d7 |
| institution | Directory Open Access Journal |
| issn | 2831-0896 2831-090X |
| language | English |
| last_indexed | 2024-04-24T23:40:20Z |
| publishDate | 2023-07-01 |
| publisher | Association of Basic Medical Sciences of Federation of Bosnia and Herzegovina |
| record_format | Article |
| series | Biomolecules & Biomedicine |
| spelling | doaj.art-9f3f3aeef5ff466b8b4a2894223767d72024-03-15T13:22:32ZengAssociation of Basic Medical Sciences of Federation of Bosnia and HerzegovinaBiomolecules & Biomedicine2831-08962831-090X2023-07-0123410.17305/bb.2022.8693A novel approach for simultaneous detection of the most common food-borne pathogens by multiplex qPCR Emir Hodzic0https://orcid.org/0000-0002-6073-4052Aida Glavinic1https://orcid.org/0000-0001-8661-7180Cara Wademan2School of Veterinary Medicine, University of California, Davis, U.S.A.Faculty of Veterinary Medicine, University of Sarajevo, Sarajevo, Bosnia and HerzegovinaSchool of Veterinary Medicine, University of California, Davis, U.S.A. Food contaminated with bacterial pathogens is a great threat to human health and food spoilage, having an impact on public health and the food industry. Research in food safety seeks to develop a practical, rapid, and sensitive detection technique for food-borne pathogens. In the past few decades, real-time quantitative polymerase chain reaction (qPCR) has been developed, and multiplex qPCR is a preferred feature. Multiplex qPCR enables the simultaneous amplification of many targets of interest in one reaction by using more than one pair of primers. In this study, we have developed and evaluated a hydrolysis (TaqMan) probe-based system for simultaneous detection of eight of the most common food-borne pathogens in a single-step procedure by multiplex qPCR. A multicolor combinational probe coding (MCPC) strategy was utilized that allows multiple fluorophores to label different probes in combinatorial manner. This strategy enabled simultaneous detection, identification, and quantification of targeted genes. The efficiency of the individual qPCR reactions for each target gene had values comparable to those established for multiplex qPCR, with detection limits of approximately < 10 copies of DNA per reaction. Pathogen load helps to predict bacteriological quality status in food products and serves to validate the efficiency of procedures to minimize or eliminate their presence, so newly developed multiplex qPCR was quantitative for each pathogen. During sample preparation, a step to concentrate the target organism from a relatively large sample size, remove all potential PCR inhibitors, and yield samples in a volume suitable for qPCR was incorporated. https://www.bjbms.org/ojs/index.php/bjbms/article/view/8693Food-borne pathogensmultiplex qPCRlive and dead bacteriasample concentration |
| spellingShingle | Emir Hodzic Aida Glavinic Cara Wademan A novel approach for simultaneous detection of the most common food-borne pathogens by multiplex qPCR Biomolecules & Biomedicine Food-borne pathogens multiplex qPCR live and dead bacteria sample concentration |
| title | A novel approach for simultaneous detection of the most common food-borne pathogens by multiplex qPCR |
| title_full | A novel approach for simultaneous detection of the most common food-borne pathogens by multiplex qPCR |
| title_fullStr | A novel approach for simultaneous detection of the most common food-borne pathogens by multiplex qPCR |
| title_full_unstemmed | A novel approach for simultaneous detection of the most common food-borne pathogens by multiplex qPCR |
| title_short | A novel approach for simultaneous detection of the most common food-borne pathogens by multiplex qPCR |
| title_sort | novel approach for simultaneous detection of the most common food borne pathogens by multiplex qpcr |
| topic | Food-borne pathogens multiplex qPCR live and dead bacteria sample concentration |
| url | https://www.bjbms.org/ojs/index.php/bjbms/article/view/8693 |
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