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Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of...
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| Zeitschriftentitel: | Applied and Environmental Microbiology |
|---|---|
| Personen und Körperschaften: | , , , , , |
| In: | Applied and Environmental Microbiology, 67, 2001, 7, S. 3122-3126 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
American Society for Microbiology
|
| Schlagwörter: |
| author_facet |
Hein, Ingeborg Lehner, Angelika Rieck, Petra Klein, Kurt Brandl, Ernst Wagner, Martin Hein, Ingeborg Lehner, Angelika Rieck, Petra Klein, Kurt Brandl, Ernst Wagner, Martin |
|---|---|
| author |
Hein, Ingeborg Lehner, Angelika Rieck, Petra Klein, Kurt Brandl, Ernst Wagner, Martin |
| spellingShingle |
Hein, Ingeborg Lehner, Angelika Rieck, Petra Klein, Kurt Brandl, Ernst Wagner, Martin Applied and Environmental Microbiology Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese Ecology Applied Microbiology and Biotechnology Food Science Biotechnology |
| author_sort |
hein, ingeborg |
| spelling |
Hein, Ingeborg Lehner, Angelika Rieck, Petra Klein, Kurt Brandl, Ernst Wagner, Martin 0099-2240 1098-5336 American Society for Microbiology Ecology Applied Microbiology and Biotechnology Food Science Biotechnology http://dx.doi.org/10.1128/aem.67.7.3122-3126.2001 <jats:title>ABSTRACT</jats:title> <jats:p> Two different real-time quantitative PCR (RTQ-PCR) approaches were applied for PCR-based quantification of <jats:italic>Staphylococcus aureus</jats:italic> cells by targeting the thermonuclease ( <jats:italic>nuc</jats:italic> ) gene. Purified DNA extracts from pure cultures of <jats:italic>S. aureus</jats:italic> were quantified in a LightCycler system using SYBR Green I. Quantification proved to be less sensitive (60 <jats:italic>nuc</jats:italic> gene copies/μl) than using a fluorigenic TaqMan probe (6 <jats:italic>nuc</jats:italic> gene copies/μl). Comparison of the LightCycler system and the well-established ABI Prism 7700 SDS with TaqMan probes revealed no statistically significant differences with respect to sensitivity and reproducibility. Application of the RTQ-PCR assay to quantify <jats:italic>S. aureus</jats:italic> cells in artificially contaminated cheeses of different types achieved sensitivities from 1.5 × 10 <jats:sup>2</jats:sup> to 6.4 × 10 <jats:sup>2</jats:sup> copies of the <jats:italic>nuc</jats:italic> gene/2 g, depending on the cheese matrix. The coefficients of correlation between log CFU and <jats:italic>nuc</jats:italic> gene copy numbers ranged from 0.979 to 0.998, thus enabling calculation of the number of CFU of <jats:italic>S. aureus</jats:italic> in cheese by performing RTQ-PCR. </jats:p> Comparison of Different Approaches To Quantify <i>Staphylococcus aureus</i> Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese Applied and Environmental Microbiology |
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10.1128/aem.67.7.3122-3126.2001 |
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Online Free |
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American Society for Microbiology, 2001 |
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Applied and Environmental Microbiology |
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| title |
Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_unstemmed |
Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_full |
Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_fullStr |
Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_full_unstemmed |
Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_short |
Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_sort |
comparison of different approaches to quantify
<i>staphylococcus aureus</i>
cells by real-time quantitative pcr and application of this technique for examination of cheese |
| topic |
Ecology Applied Microbiology and Biotechnology Food Science Biotechnology |
| url |
http://dx.doi.org/10.1128/aem.67.7.3122-3126.2001 |
| publishDate |
2001 |
| physical |
3122-3126 |
| description |
<jats:title>ABSTRACT</jats:title>
<jats:p>
Two different real-time quantitative PCR (RTQ-PCR) approaches were applied for PCR-based quantification of
<jats:italic>Staphylococcus aureus</jats:italic>
cells by targeting the thermonuclease (
<jats:italic>nuc</jats:italic>
) gene. Purified DNA extracts from pure cultures of
<jats:italic>S. aureus</jats:italic>
were quantified in a LightCycler system using SYBR Green I. Quantification proved to be less sensitive (60
<jats:italic>nuc</jats:italic>
gene copies/μl) than using a fluorigenic TaqMan probe (6
<jats:italic>nuc</jats:italic>
gene copies/μl). Comparison of the LightCycler system and the well-established ABI Prism 7700 SDS with TaqMan probes revealed no statistically significant differences with respect to sensitivity and reproducibility. Application of the RTQ-PCR assay to quantify
<jats:italic>S. aureus</jats:italic>
cells in artificially contaminated cheeses of different types achieved sensitivities from 1.5 × 10
<jats:sup>2</jats:sup>
to 6.4 × 10
<jats:sup>2</jats:sup>
copies of the
<jats:italic>nuc</jats:italic>
gene/2 g, depending on the cheese matrix. The coefficients of correlation between log CFU and
<jats:italic>nuc</jats:italic>
gene copy numbers ranged from 0.979 to 0.998, thus enabling calculation of the number of CFU of
<jats:italic>S. aureus</jats:italic>
in cheese by performing RTQ-PCR.
</jats:p> |
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| author | Hein, Ingeborg, Lehner, Angelika, Rieck, Petra, Klein, Kurt, Brandl, Ernst, Wagner, Martin |
| author_facet | Hein, Ingeborg, Lehner, Angelika, Rieck, Petra, Klein, Kurt, Brandl, Ernst, Wagner, Martin, Hein, Ingeborg, Lehner, Angelika, Rieck, Petra, Klein, Kurt, Brandl, Ernst, Wagner, Martin |
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| container_issue | 7 |
| container_start_page | 3122 |
| container_title | Applied and Environmental Microbiology |
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| description | <jats:title>ABSTRACT</jats:title> <jats:p> Two different real-time quantitative PCR (RTQ-PCR) approaches were applied for PCR-based quantification of <jats:italic>Staphylococcus aureus</jats:italic> cells by targeting the thermonuclease ( <jats:italic>nuc</jats:italic> ) gene. Purified DNA extracts from pure cultures of <jats:italic>S. aureus</jats:italic> were quantified in a LightCycler system using SYBR Green I. Quantification proved to be less sensitive (60 <jats:italic>nuc</jats:italic> gene copies/μl) than using a fluorigenic TaqMan probe (6 <jats:italic>nuc</jats:italic> gene copies/μl). Comparison of the LightCycler system and the well-established ABI Prism 7700 SDS with TaqMan probes revealed no statistically significant differences with respect to sensitivity and reproducibility. Application of the RTQ-PCR assay to quantify <jats:italic>S. aureus</jats:italic> cells in artificially contaminated cheeses of different types achieved sensitivities from 1.5 × 10 <jats:sup>2</jats:sup> to 6.4 × 10 <jats:sup>2</jats:sup> copies of the <jats:italic>nuc</jats:italic> gene/2 g, depending on the cheese matrix. The coefficients of correlation between log CFU and <jats:italic>nuc</jats:italic> gene copy numbers ranged from 0.979 to 0.998, thus enabling calculation of the number of CFU of <jats:italic>S. aureus</jats:italic> in cheese by performing RTQ-PCR. </jats:p> |
| doi_str_mv | 10.1128/aem.67.7.3122-3126.2001 |
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| imprint | American Society for Microbiology, 2001 |
| imprint_str_mv | American Society for Microbiology, 2001 |
| institution | DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229 |
| issn | 1098-5336, 0099-2240 |
| issn_str_mv | 1098-5336, 0099-2240 |
| language | English |
| last_indexed | 2024-03-01T17:14:53.87Z |
| match_str | hein2001comparisonofdifferentapproachestoquantifystaphylococcusaureuscellsbyrealtimequantitativepcrandapplicationofthistechniqueforexaminationofcheese |
| mega_collection | American Society for Microbiology (CrossRef) |
| physical | 3122-3126 |
| publishDate | 2001 |
| publishDateSort | 2001 |
| publisher | American Society for Microbiology |
| record_format | ai |
| recordtype | ai |
| series | Applied and Environmental Microbiology |
| source_id | 49 |
| spelling | Hein, Ingeborg Lehner, Angelika Rieck, Petra Klein, Kurt Brandl, Ernst Wagner, Martin 0099-2240 1098-5336 American Society for Microbiology Ecology Applied Microbiology and Biotechnology Food Science Biotechnology http://dx.doi.org/10.1128/aem.67.7.3122-3126.2001 <jats:title>ABSTRACT</jats:title> <jats:p> Two different real-time quantitative PCR (RTQ-PCR) approaches were applied for PCR-based quantification of <jats:italic>Staphylococcus aureus</jats:italic> cells by targeting the thermonuclease ( <jats:italic>nuc</jats:italic> ) gene. Purified DNA extracts from pure cultures of <jats:italic>S. aureus</jats:italic> were quantified in a LightCycler system using SYBR Green I. Quantification proved to be less sensitive (60 <jats:italic>nuc</jats:italic> gene copies/μl) than using a fluorigenic TaqMan probe (6 <jats:italic>nuc</jats:italic> gene copies/μl). Comparison of the LightCycler system and the well-established ABI Prism 7700 SDS with TaqMan probes revealed no statistically significant differences with respect to sensitivity and reproducibility. Application of the RTQ-PCR assay to quantify <jats:italic>S. aureus</jats:italic> cells in artificially contaminated cheeses of different types achieved sensitivities from 1.5 × 10 <jats:sup>2</jats:sup> to 6.4 × 10 <jats:sup>2</jats:sup> copies of the <jats:italic>nuc</jats:italic> gene/2 g, depending on the cheese matrix. The coefficients of correlation between log CFU and <jats:italic>nuc</jats:italic> gene copy numbers ranged from 0.979 to 0.998, thus enabling calculation of the number of CFU of <jats:italic>S. aureus</jats:italic> in cheese by performing RTQ-PCR. </jats:p> Comparison of Different Approaches To Quantify <i>Staphylococcus aureus</i> Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese Applied and Environmental Microbiology |
| spellingShingle | Hein, Ingeborg, Lehner, Angelika, Rieck, Petra, Klein, Kurt, Brandl, Ernst, Wagner, Martin, Applied and Environmental Microbiology, Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese, Ecology, Applied Microbiology and Biotechnology, Food Science, Biotechnology |
| title | Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_full | Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_fullStr | Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_full_unstemmed | Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_short | Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| title_sort | comparison of different approaches to quantify <i>staphylococcus aureus</i> cells by real-time quantitative pcr and application of this technique for examination of cheese |
| title_unstemmed | Comparison of Different Approaches To Quantify Staphylococcus aureus Cells by Real-Time Quantitative PCR and Application of This Technique for Examination of Cheese |
| topic | Ecology, Applied Microbiology and Biotechnology, Food Science, Biotechnology |
| url | http://dx.doi.org/10.1128/aem.67.7.3122-3126.2001 |