Staff View: Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases

Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases

The case of foodborne disease by pathogenic bacteria is an extraordinary event whose source and route of spread occur quickly. Bacillus cereus and Listeria monocytogenes are included in the gram-positive bacteria that cause food poisoning infections in humans. In previous studies, the conditions and...

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Main Authors:	Muktiningsih Nurjayadi, Muktiningsih Nurjayadi, Irvan Maulana, Irvan Maulana, Nabila Alya Pramudiyasih, Nabila Alya Pramudiyasih, Ratna Nur Kusumawati, Ratna Nur Kusumawati, Maharaniaska Azzahra, Maharaniaska Azzahra, Niken Kurnia Liman, Niken Kurnia Liman, Muhammad Arkent Sangkara, Muhammad Arkent Sangkara, Esnawan Wibisono, Esnawan Wibisono, Fera Kurniadewi, Fera Kurniadewi, Vira Saamia, Vira Saamia, Dwi Anna Oktaviani Saputro, Dwi Anna Oktaviani Saputro, I. Made Wiranatha, I. Made Wiranatha, El-Enshasy, Hesham Ali
Format:	Conference or Workshop Item
Published:	2023
Subjects:	TP Chemical technology
Online Access:	http://eprints.utm.my/108163/ http://dx.doi.org/10.1063/5.0126315
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id	my.utm.108163
record_format	eprints
spelling	my.utm.1081632024-10-22T07:38:25Z http://eprints.utm.my/108163/ Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases Muktiningsih Nurjayadi, Muktiningsih Nurjayadi Irvan Maulana, Irvan Maulana Nabila Alya Pramudiyasih, Nabila Alya Pramudiyasih Ratna Nur Kusumawati, Ratna Nur Kusumawati Maharaniaska Azzahra, Maharaniaska Azzahra Niken Kurnia Liman, Niken Kurnia Liman Muhammad Arkent Sangkara, Muhammad Arkent Sangkara Esnawan Wibisono, Esnawan Wibisono Fera Kurniadewi, Fera Kurniadewi Vira Saamia, Vira Saamia Dwi Anna Oktaviani Saputro, Dwi Anna Oktaviani Saputro I. Made Wiranatha, I. Made Wiranatha El-Enshasy, Hesham Ali TP Chemical technology The case of foodborne disease by pathogenic bacteria is an extraordinary event whose source and route of spread occur quickly. Bacillus cereus and Listeria monocytogenes are included in the gram-positive bacteria that cause food poisoning infections in humans. In previous studies, the conditions and reaction formulas for the prototype rapid kit detections of foodborne pathogens were obtained using the Real-Time PCR method. This research validates the conditions and reaction formulas in the prior stage to obtain a consistent and reproducible prototype product. Validation of the isolation stage of Bacillus cereus and Listeria monocytogenes bacteria was carried out by culturing bacterial cultures at aeration conditions of 250 rpm and a temperature of 37oC. A total of three mL of the culture was centrifuged at 5000 rpm and yielded 0.03 and 0.02 grams of B. cereus and L. monocytogenes cells, respectively. The results of the isolation of genomic DNA with Qiaprep were 200 µL with the amount of DNA 10600-11700 nanograms; the concentration was 23-58.5 ng/µL, and the purity was 1.809-2.084. Furthermore, the resulting it is used as a template for the PCR process. The validation of the PCR stage of B. cereus and L. monocytogenes at a temperature of 60oC and template concentration of 50-60 ng resulted in an amplicon of both bacteria. The cyt-k gene B. cereus and hly L. monocytogenes are measuring respectively 114 bp and 158 bp. Based on the data obtained, it can be concluded that the validation of the isolation and PCR stages has been successfully carried out by producing data consistent with previous studies. The development of a detection kit prototype is then revealed to the Real-Time PCR stage and laboratory-scale trials for artificially contaminated food samples and food samples from products on the market. 2023 Conference or Workshop Item PeerReviewed Muktiningsih Nurjayadi, Muktiningsih Nurjayadi and Irvan Maulana, Irvan Maulana and Nabila Alya Pramudiyasih, Nabila Alya Pramudiyasih and Ratna Nur Kusumawati, Ratna Nur Kusumawati and Maharaniaska Azzahra, Maharaniaska Azzahra and Niken Kurnia Liman, Niken Kurnia Liman and Muhammad Arkent Sangkara, Muhammad Arkent Sangkara and Esnawan Wibisono, Esnawan Wibisono and Fera Kurniadewi, Fera Kurniadewi and Vira Saamia, Vira Saamia and Dwi Anna Oktaviani Saputro, Dwi Anna Oktaviani Saputro and I. Made Wiranatha, I. Made Wiranatha and El-Enshasy, Hesham Ali (2023) Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases. In: 8th International Conference on Mathematics, Science and Education, ICMSE 2021, 5 October 2021-6 October 2021, Virtual, Online, Semarang, Indonesia. http://dx.doi.org/10.1063/5.0126315
institution	Universiti Teknologi Malaysia
building	UTM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Teknologi Malaysia
content_source	UTM Institutional Repository
url_provider	http://eprints.utm.my/
topic	TP Chemical technology
spellingShingle	TP Chemical technology Muktiningsih Nurjayadi, Muktiningsih Nurjayadi Irvan Maulana, Irvan Maulana Nabila Alya Pramudiyasih, Nabila Alya Pramudiyasih Ratna Nur Kusumawati, Ratna Nur Kusumawati Maharaniaska Azzahra, Maharaniaska Azzahra Niken Kurnia Liman, Niken Kurnia Liman Muhammad Arkent Sangkara, Muhammad Arkent Sangkara Esnawan Wibisono, Esnawan Wibisono Fera Kurniadewi, Fera Kurniadewi Vira Saamia, Vira Saamia Dwi Anna Oktaviani Saputro, Dwi Anna Oktaviani Saputro I. Made Wiranatha, I. Made Wiranatha El-Enshasy, Hesham Ali Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases
description	The case of foodborne disease by pathogenic bacteria is an extraordinary event whose source and route of spread occur quickly. Bacillus cereus and Listeria monocytogenes are included in the gram-positive bacteria that cause food poisoning infections in humans. In previous studies, the conditions and reaction formulas for the prototype rapid kit detections of foodborne pathogens were obtained using the Real-Time PCR method. This research validates the conditions and reaction formulas in the prior stage to obtain a consistent and reproducible prototype product. Validation of the isolation stage of Bacillus cereus and Listeria monocytogenes bacteria was carried out by culturing bacterial cultures at aeration conditions of 250 rpm and a temperature of 37oC. A total of three mL of the culture was centrifuged at 5000 rpm and yielded 0.03 and 0.02 grams of B. cereus and L. monocytogenes cells, respectively. The results of the isolation of genomic DNA with Qiaprep were 200 µL with the amount of DNA 10600-11700 nanograms; the concentration was 23-58.5 ng/µL, and the purity was 1.809-2.084. Furthermore, the resulting it is used as a template for the PCR process. The validation of the PCR stage of B. cereus and L. monocytogenes at a temperature of 60oC and template concentration of 50-60 ng resulted in an amplicon of both bacteria. The cyt-k gene B. cereus and hly L. monocytogenes are measuring respectively 114 bp and 158 bp. Based on the data obtained, it can be concluded that the validation of the isolation and PCR stages has been successfully carried out by producing data consistent with previous studies. The development of a detection kit prototype is then revealed to the Real-Time PCR stage and laboratory-scale trials for artificially contaminated food samples and food samples from products on the market.
format	Conference or Workshop Item
author	Muktiningsih Nurjayadi, Muktiningsih Nurjayadi Irvan Maulana, Irvan Maulana Nabila Alya Pramudiyasih, Nabila Alya Pramudiyasih Ratna Nur Kusumawati, Ratna Nur Kusumawati Maharaniaska Azzahra, Maharaniaska Azzahra Niken Kurnia Liman, Niken Kurnia Liman Muhammad Arkent Sangkara, Muhammad Arkent Sangkara Esnawan Wibisono, Esnawan Wibisono Fera Kurniadewi, Fera Kurniadewi Vira Saamia, Vira Saamia Dwi Anna Oktaviani Saputro, Dwi Anna Oktaviani Saputro I. Made Wiranatha, I. Made Wiranatha El-Enshasy, Hesham Ali
author_facet	Muktiningsih Nurjayadi, Muktiningsih Nurjayadi Irvan Maulana, Irvan Maulana Nabila Alya Pramudiyasih, Nabila Alya Pramudiyasih Ratna Nur Kusumawati, Ratna Nur Kusumawati Maharaniaska Azzahra, Maharaniaska Azzahra Niken Kurnia Liman, Niken Kurnia Liman Muhammad Arkent Sangkara, Muhammad Arkent Sangkara Esnawan Wibisono, Esnawan Wibisono Fera Kurniadewi, Fera Kurniadewi Vira Saamia, Vira Saamia Dwi Anna Oktaviani Saputro, Dwi Anna Oktaviani Saputro I. Made Wiranatha, I. Made Wiranatha El-Enshasy, Hesham Ali
author_sort	Muktiningsih Nurjayadi, Muktiningsih Nurjayadi
title	Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases
title_short	Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases
title_full	Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases
title_fullStr	Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases
title_full_unstemmed	Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases
title_sort	isolation and pcr validation of bacillus cereus and listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases
publishDate	2023
url	http://eprints.utm.my/108163/ http://dx.doi.org/10.1063/5.0126315
_version_	1814043615996936192
score	13.211869

Isolation and PCR validation of Bacillus cereus and Listeria monocytogenes bacteria for the development of prototype kit detection of foodborne pathogens diseases

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