Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification of <i>Listeria monocytogenes</i> in Meat Products
In this study a real-time PCR method has been developed for the specific quantification of the foodborne pathogen <i>Listeria monocytogenes</i> on meat products through the gene <i>hlyA</i>. The PCR was combined with a matrix lysis that allowed the obtaining of the microorgan...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-03-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/10/4/735 |
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| author | Mirian Labrador Carlota Giménez-Rota Carmen Rota |
| author_facet | Mirian Labrador Carlota Giménez-Rota Carmen Rota |
| author_sort | Mirian Labrador |
| collection | DOAJ |
| description | In this study a real-time PCR method has been developed for the specific quantification of the foodborne pathogen <i>Listeria monocytogenes</i> on meat products through the gene <i>hlyA</i>. The PCR was combined with a matrix lysis that allowed the obtaining of the microorganisms without sample dilution and the elimination the PCR inhibitors from dry-cured ham. The qPCR method calibration curve had an efficiency of 100.4%, limits of detection and quantification were 30.1 ± 6.2 CFU/g which is under the legal limit of <i>L. monocytogenes</i> in ready-to-eat products, and an analytical variability <0.25 log <i>hlyA</i> gene copies/reaction. The analysis was performed simultaneously with the reference method ISO 11290-2. The comparison of the qPCR-matrix lysis results with the reference method showed an excellent correspondence, with a relative accuracy between 95.83–105.20%. Finally, the method was applied to commercial derived meat samples and the pathogen was quantified in one of the commercial samples assayed in 69.1 ± 13.9 CFU/g while the reference method did not quantify it. The optimized qPCR showed higher precision and sensitivity than the reference method at low concentrations of the microorganism in a shorter time. Therefore, qPCR-matrix lysis shows a potential application in the meat industry for <i>L. monocytogenes</i> routine control. |
| first_indexed | 2024-03-10T12:46:48Z |
| format | Article |
| id | doaj.art-b821316720c9428184ca1ff5e6d1381b |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-10T12:46:48Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-b821316720c9428184ca1ff5e6d1381b2023-11-21T13:26:55ZengMDPI AGFoods2304-81582021-03-0110473510.3390/foods10040735Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification of <i>Listeria monocytogenes</i> in Meat ProductsMirian Labrador0Carlota Giménez-Rota1Carmen Rota2Departamento de Producción Animal y Ciencia de los Alimentos, Facultad de Veterinaria, Instituto Agroalimentario de Aragón -IA2-, Universidad de Zaragoza-CITA, C/Miguel Servet 177, 50013 Zaragoza, SpainFacultad de Farmacia, Universidad Complutense de Madrid, Plaza Ramón y Cajal s/n, 28040 Madrid, SpainDepartamento de Producción Animal y Ciencia de los Alimentos, Facultad de Veterinaria, Instituto Agroalimentario de Aragón -IA2-, Universidad de Zaragoza-CITA, C/Miguel Servet 177, 50013 Zaragoza, SpainIn this study a real-time PCR method has been developed for the specific quantification of the foodborne pathogen <i>Listeria monocytogenes</i> on meat products through the gene <i>hlyA</i>. The PCR was combined with a matrix lysis that allowed the obtaining of the microorganisms without sample dilution and the elimination the PCR inhibitors from dry-cured ham. The qPCR method calibration curve had an efficiency of 100.4%, limits of detection and quantification were 30.1 ± 6.2 CFU/g which is under the legal limit of <i>L. monocytogenes</i> in ready-to-eat products, and an analytical variability <0.25 log <i>hlyA</i> gene copies/reaction. The analysis was performed simultaneously with the reference method ISO 11290-2. The comparison of the qPCR-matrix lysis results with the reference method showed an excellent correspondence, with a relative accuracy between 95.83–105.20%. Finally, the method was applied to commercial derived meat samples and the pathogen was quantified in one of the commercial samples assayed in 69.1 ± 13.9 CFU/g while the reference method did not quantify it. The optimized qPCR showed higher precision and sensitivity than the reference method at low concentrations of the microorganism in a shorter time. Therefore, qPCR-matrix lysis shows a potential application in the meat industry for <i>L. monocytogenes</i> routine control.https://www.mdpi.com/2304-8158/10/4/735real time PCRquantification<i>Listeria monocytogenes</i>meat productsmatrix lysis |
| spellingShingle | Mirian Labrador Carlota Giménez-Rota Carmen Rota Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification of <i>Listeria monocytogenes</i> in Meat Products Foods real time PCR quantification <i>Listeria monocytogenes</i> meat products matrix lysis |
| title | Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification of <i>Listeria monocytogenes</i> in Meat Products |
| title_full | Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification of <i>Listeria monocytogenes</i> in Meat Products |
| title_fullStr | Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification of <i>Listeria monocytogenes</i> in Meat Products |
| title_full_unstemmed | Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification of <i>Listeria monocytogenes</i> in Meat Products |
| title_short | Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification of <i>Listeria monocytogenes</i> in Meat Products |
| title_sort | real time pcr method combined with a matrix lysis procedure for the quantification of i listeria monocytogenes i in meat products |
| topic | real time PCR quantification <i>Listeria monocytogenes</i> meat products matrix lysis |
| url | https://www.mdpi.com/2304-8158/10/4/735 |
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