Primer concentration and Pre-denaturation Time Effect on cyt-K Bacillus cereus Detection using Real-Time PCR Method
Foodborne disease is a global threat that can affect all sections of society, both in developed or developing countries. Bacillus cereus is a Gram-positive bacteria that can cause food poisoning disease in humans. [2] Real-Time PCR detection method is one of the molecular marker methods that has bee...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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EDP Sciences
2023-01-01
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| Series: | E3S Web of Conferences |
| Online Access: | https://www.e3s-conferences.org/articles/e3sconf/pdf/2023/37/e3sconf_icosmed2023_04011.pdf |
| _version_ | 1797775948920127488 |
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| author | Azzahra M. Nurjayadi M. Pramudiyasih N. A. Kusumawati R. N. Maulana I. Declan J. L. Putri G. I. Juliansyah D. A. Krisdawati I. Kurniadewi F. Kartika I. R. Sukmawati D. Nastassya L. Saamia V. Oktaviani D.A.S. Wiranatha M. Ali El-Enshasy H. Abomoelak B. |
| author_facet | Azzahra M. Nurjayadi M. Pramudiyasih N. A. Kusumawati R. N. Maulana I. Declan J. L. Putri G. I. Juliansyah D. A. Krisdawati I. Kurniadewi F. Kartika I. R. Sukmawati D. Nastassya L. Saamia V. Oktaviani D.A.S. Wiranatha M. Ali El-Enshasy H. Abomoelak B. |
| author_sort | Azzahra M. |
| collection | DOAJ |
| description | Foodborne disease is a global threat that can affect all sections of society, both in developed or developing countries. Bacillus cereus is a Gram-positive bacteria that can cause food poisoning disease in humans. [2] Real-Time PCR detection method is one of the molecular marker methods that has been widely recognized as a fast, reliable, sensitive and specific detection tool for detecting pathogenic bacteria. In previous studies, the optimum condition and formulas applied for cyt-K 2 primer pairs have been obtained using Real-Time PCR. The purpose of this study is to find out the best conditions work of the primer pair cyt-K Bacillus cereus on detecting bacteria target using variations of pre-denaturation time and primer concentration with Real-Time PCR method. The annealing temperature used for PCR is at 60°C with sample concentration 50 ng/µL of B. cereus. Real-time PCR detection of variations in pre-denaturation time and primer concentration obtained the best conditions for primer pair cyt-K work at minute 4 with a primer concentration of 10 pmol and successfully amplifying the target by producing a Ct value of B. cereus at 13.04. Based on the results of the study, the primer pair cyt-K were reproducible in detecting the target gene and in the further step, this research can be continued to developed a prototype detection kit for foodborne pathogen bacteria using Real-Time PCR method. |
| first_indexed | 2024-03-12T22:43:04Z |
| format | Article |
| id | doaj.art-3646d6e1dab047a6b25bc207cd3e24c2 |
| institution | Directory Open Access Journal |
| issn | 2267-1242 |
| language | English |
| last_indexed | 2024-03-12T22:43:04Z |
| publishDate | 2023-01-01 |
| publisher | EDP Sciences |
| record_format | Article |
| series | E3S Web of Conferences |
| spelling | doaj.art-3646d6e1dab047a6b25bc207cd3e24c22023-07-21T09:33:17ZengEDP SciencesE3S Web of Conferences2267-12422023-01-014000401110.1051/e3sconf/202340004011e3sconf_icosmed2023_04011Primer concentration and Pre-denaturation Time Effect on cyt-K Bacillus cereus Detection using Real-Time PCR MethodAzzahra M.0Nurjayadi M.1Pramudiyasih N. A.2Kusumawati R. N.3Maulana I.4Declan J. L.5Putri G. I.6Juliansyah D. A.7Krisdawati I.8Kurniadewi F.9Kartika I. R.10Sukmawati D.11Nastassya L.12Saamia V.13Oktaviani D.A.S.Wiranatha M.14Ali El-Enshasy H.15Abomoelak B.16Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariDepartment of Biology, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj’ariCenter for Agricultural Quarantine Standards Test, Jl. Pemuda RawamangunCenter Forensic Laboratory of the Criminal InvestigationCenter Forensic Laboratory of the Criminal InvestigationInstitute of Bioproduct Development (IBD), Universiti Teknologi Malaysia (UTM)Arnold Palmer Hospital Paediatric Speciality Diagnostic LaboratoryFoodborne disease is a global threat that can affect all sections of society, both in developed or developing countries. Bacillus cereus is a Gram-positive bacteria that can cause food poisoning disease in humans. [2] Real-Time PCR detection method is one of the molecular marker methods that has been widely recognized as a fast, reliable, sensitive and specific detection tool for detecting pathogenic bacteria. In previous studies, the optimum condition and formulas applied for cyt-K 2 primer pairs have been obtained using Real-Time PCR. The purpose of this study is to find out the best conditions work of the primer pair cyt-K Bacillus cereus on detecting bacteria target using variations of pre-denaturation time and primer concentration with Real-Time PCR method. The annealing temperature used for PCR is at 60°C with sample concentration 50 ng/µL of B. cereus. Real-time PCR detection of variations in pre-denaturation time and primer concentration obtained the best conditions for primer pair cyt-K work at minute 4 with a primer concentration of 10 pmol and successfully amplifying the target by producing a Ct value of B. cereus at 13.04. Based on the results of the study, the primer pair cyt-K were reproducible in detecting the target gene and in the further step, this research can be continued to developed a prototype detection kit for foodborne pathogen bacteria using Real-Time PCR method.https://www.e3s-conferences.org/articles/e3sconf/pdf/2023/37/e3sconf_icosmed2023_04011.pdf |
| spellingShingle | Azzahra M. Nurjayadi M. Pramudiyasih N. A. Kusumawati R. N. Maulana I. Declan J. L. Putri G. I. Juliansyah D. A. Krisdawati I. Kurniadewi F. Kartika I. R. Sukmawati D. Nastassya L. Saamia V. Oktaviani D.A.S. Wiranatha M. Ali El-Enshasy H. Abomoelak B. Primer concentration and Pre-denaturation Time Effect on cyt-K Bacillus cereus Detection using Real-Time PCR Method E3S Web of Conferences |
| title | Primer concentration and Pre-denaturation Time Effect on cyt-K Bacillus cereus Detection using Real-Time PCR Method |
| title_full | Primer concentration and Pre-denaturation Time Effect on cyt-K Bacillus cereus Detection using Real-Time PCR Method |
| title_fullStr | Primer concentration and Pre-denaturation Time Effect on cyt-K Bacillus cereus Detection using Real-Time PCR Method |
| title_full_unstemmed | Primer concentration and Pre-denaturation Time Effect on cyt-K Bacillus cereus Detection using Real-Time PCR Method |
| title_short | Primer concentration and Pre-denaturation Time Effect on cyt-K Bacillus cereus Detection using Real-Time PCR Method |
| title_sort | primer concentration and pre denaturation time effect on cyt k bacillus cereus detection using real time pcr method |
| url | https://www.e3s-conferences.org/articles/e3sconf/pdf/2023/37/e3sconf_icosmed2023_04011.pdf |
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