Rapid Detection of Foodborne Pathogen Bacteria Vibrio parahaemolyticus in Seafood Using Gene ToxR with Real-Time Polymerase Chain Reaction Method
Cases of food poisoning often occur due to food contamination caused by pathogenic bacteria. One of the pathogenic bacteria is Vibrio parahaemolyticus which is found in seafood. Thus, a fast, accurate and specific detection method is needed. The purpose of this study was to quickly detect Vibrio...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Indonesian Society for Microbiology
2023-05-01
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| Series: | Microbiology Indonesia |
| Online Access: | https://jurnal.permi.or.id/index.php/mionline/article/view/1038 |
| _version_ | 1797826562168455168 |
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| author | Ismaya Krisdawati Muktiningsih Nurjayadi Jefferson Lynford Declan Gladys Indira Putri Dandy Akbar Juliansyah Maharani Azka Azzahra Irvan Maulana Irma Ratna Kartika Vira Saamia Dwi Ana Oktaviani I Made Wiranatha Hesham Ali Al-Enshashy |
| author_facet | Ismaya Krisdawati Muktiningsih Nurjayadi Jefferson Lynford Declan Gladys Indira Putri Dandy Akbar Juliansyah Maharani Azka Azzahra Irvan Maulana Irma Ratna Kartika Vira Saamia Dwi Ana Oktaviani I Made Wiranatha Hesham Ali Al-Enshashy |
| author_sort | Ismaya Krisdawati |
| collection | DOAJ |
| description |
Cases of food poisoning often occur due to food contamination caused by pathogenic bacteria. One of the
pathogenic bacteria is Vibrio parahaemolyticus which is found in seafood. Thus, a fast, accurate and specific
detection method is needed. The purpose of this study was to quickly detect Vibrio parahaemolyticus bacteria
in seafood samples targeting the ToxR gene using Real Time PCR. In a previous study, gradient PCR was used
to optimize ideal annealing temperature ranges from 53-62°C and revealed that 58°C produced the best
outcomes for the ToxR primer with a size of 171 base pairs. Real-Time PCR was utilized to amplify, specify,
and test for sensitivity under the ideal conditions from the PCR Gradient. The confirmation results show that
the primer pairs could amplify ToxR of Vibrio parahaemolyticus with the amount of concentration as much as
50 ng/µL with Ct 10,69 and 10,32 and melting curve at temperature 82,18°C and 82,23°C. This primer pair
can also distinguish non-target bacteria with different Ct and melting curve temperature. The sensitivity assay
for this primer can amplify DNA templates at concentration 0,0032 ng/µL. Shrimp samples that are
contaminated artificially can still be detected at Ct 13,02 and Ct 13,09. Based on these results, it can be
concluded that Real Time PCR with ToxR primer can be applied to develop a detection kit for Vibrio
parahaemolyticus in seafood.
|
| first_indexed | 2024-04-09T12:34:22Z |
| format | Article |
| id | doaj.art-7f7634f7bc454ca5aa6e8e1d017c535a |
| institution | Directory Open Access Journal |
| issn | 1978-3477 2087-8575 |
| language | English |
| last_indexed | 2024-04-09T12:34:22Z |
| publishDate | 2023-05-01 |
| publisher | Indonesian Society for Microbiology |
| record_format | Article |
| series | Microbiology Indonesia |
| spelling | doaj.art-7f7634f7bc454ca5aa6e8e1d017c535a2023-05-15T16:23:55ZengIndonesian Society for MicrobiologyMicrobiology Indonesia1978-34772087-85752023-05-01171Rapid Detection of Foodborne Pathogen Bacteria Vibrio parahaemolyticus in Seafood Using Gene ToxR with Real-Time Polymerase Chain Reaction MethodIsmaya Krisdawati0Muktiningsih Nurjayadi1Jefferson Lynford Declan2Gladys Indira Putri3Dandy Akbar Juliansyah4Maharani Azka Azzahra5Irvan Maulana6Irma Ratna Kartika7Vira Saamia8Dwi Ana Oktaviani9I Made Wiranatha10Hesham Ali Al-Enshashy11Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj'ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, IndonesiaDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj'ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia; Research Center for Detection of Pathogenic Bacteria, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Universitas Negeri Jakarta, Jl. Rawamangun Muka, Jakarta Timur 13220, IndonesiaDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj'ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia; Research Center for Detection of Pathogenic Bacteria, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Universitas Negeri Jakarta, Jl. Rawamangun Muka, Jakarta Timur 13220, IndonesiaDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj'ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia; Research Center for Detection of Pathogenic Bacteria, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Universitas Negeri Jakarta, Jl. Rawamangun Muka, Jakarta Timur 13220, IndonesiaDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj'ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia; Research Center for Detection of Pathogenic Bacteria, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Universitas Negeri Jakarta, Jl. Rawamangun Muka, Jakarta Timur 13220, IndonesiaDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj'ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia; Research Center for Detection of Pathogenic Bacteria, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Universitas Negeri Jakarta, Jl. Rawamangun Muka, Jakarta Timur 13220, IndonesiaDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj'ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia; Research Center for Detection of Pathogenic Bacteria, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Universitas Negeri Jakarta, Jl. Rawamangun Muka, Jakarta Timur 13220, IndonesiaDepartment of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj'ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia; Research Center for Detection of Pathogenic Bacteria, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Universitas Negeri Jakarta, Jl. Rawamangun Muka, Jakarta Timur 13220, IndonesiaCenter Forensic Laboratory of the Criminal Investigation, Police of the Republic of Indonesia, Cipambuan Babakan Madang, Bogor, 1681, IndonesiaCenter Forensic Laboratory of the Criminal Investigation, Police of the Republic of Indonesia, Cipambuan Babakan Madang, Bogor, 1681, IndonesiaCenter Forensic Laboratory of the Criminal Investigation, Police of the Republic of Indonesia, Cipambuan Babakan Madang, Bogor, 1681, IndonesiaInstitute of Bioproduct Development, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia; School of Chemical and Energy Engineering, Faculty of Engineering, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia; City of Scientific Research and Technology Applications, New Burg Al Arab, Alexandria, Egypt Cases of food poisoning often occur due to food contamination caused by pathogenic bacteria. One of the pathogenic bacteria is Vibrio parahaemolyticus which is found in seafood. Thus, a fast, accurate and specific detection method is needed. The purpose of this study was to quickly detect Vibrio parahaemolyticus bacteria in seafood samples targeting the ToxR gene using Real Time PCR. In a previous study, gradient PCR was used to optimize ideal annealing temperature ranges from 53-62°C and revealed that 58°C produced the best outcomes for the ToxR primer with a size of 171 base pairs. Real-Time PCR was utilized to amplify, specify, and test for sensitivity under the ideal conditions from the PCR Gradient. The confirmation results show that the primer pairs could amplify ToxR of Vibrio parahaemolyticus with the amount of concentration as much as 50 ng/µL with Ct 10,69 and 10,32 and melting curve at temperature 82,18°C and 82,23°C. This primer pair can also distinguish non-target bacteria with different Ct and melting curve temperature. The sensitivity assay for this primer can amplify DNA templates at concentration 0,0032 ng/µL. Shrimp samples that are contaminated artificially can still be detected at Ct 13,02 and Ct 13,09. Based on these results, it can be concluded that Real Time PCR with ToxR primer can be applied to develop a detection kit for Vibrio parahaemolyticus in seafood. https://jurnal.permi.or.id/index.php/mionline/article/view/1038 |
| spellingShingle | Ismaya Krisdawati Muktiningsih Nurjayadi Jefferson Lynford Declan Gladys Indira Putri Dandy Akbar Juliansyah Maharani Azka Azzahra Irvan Maulana Irma Ratna Kartika Vira Saamia Dwi Ana Oktaviani I Made Wiranatha Hesham Ali Al-Enshashy Rapid Detection of Foodborne Pathogen Bacteria Vibrio parahaemolyticus in Seafood Using Gene ToxR with Real-Time Polymerase Chain Reaction Method Microbiology Indonesia |
| title | Rapid Detection of Foodborne Pathogen Bacteria Vibrio parahaemolyticus in Seafood Using Gene ToxR with Real-Time Polymerase Chain Reaction Method |
| title_full | Rapid Detection of Foodborne Pathogen Bacteria Vibrio parahaemolyticus in Seafood Using Gene ToxR with Real-Time Polymerase Chain Reaction Method |
| title_fullStr | Rapid Detection of Foodborne Pathogen Bacteria Vibrio parahaemolyticus in Seafood Using Gene ToxR with Real-Time Polymerase Chain Reaction Method |
| title_full_unstemmed | Rapid Detection of Foodborne Pathogen Bacteria Vibrio parahaemolyticus in Seafood Using Gene ToxR with Real-Time Polymerase Chain Reaction Method |
| title_short | Rapid Detection of Foodborne Pathogen Bacteria Vibrio parahaemolyticus in Seafood Using Gene ToxR with Real-Time Polymerase Chain Reaction Method |
| title_sort | rapid detection of foodborne pathogen bacteria vibrio parahaemolyticus in seafood using gene toxr with real time polymerase chain reaction method |
| url | https://jurnal.permi.or.id/index.php/mionline/article/view/1038 |
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