The Potential of htrA Gene to Identify Enterococcus faecalis using real-time Polymerase Chain Reaction

Irma Ratna Kartika; Grace Grace; Ananda Indah Putri Sihombing; Agus Setiawan; Anisa Fitriyanti; Helzi Angelina; Gusti Angieta; Puan Aqila Azizah; Royna Rahma Musie; Jefferson Lynford Declan; Gladys Indira Putri; Siti Fatimah; Ayu Berkahingrum; Muktiningsih Nurjayadi; Fera Kurniadewi; Bassam Abomoelak; Hesham A. El Enshasy

doi:10.2991/978-94-6463-624-6_12

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The Potential of htrA Gene to Identify Enterococcus faecalis using real-time Polymerase Chain Reaction

Authors

Irma Ratna Kartika¹^{, 2}^{, *}, Grace Grace¹^{, 2}, Ananda Indah Putri Sihombing¹^{, 2}, Agus Setiawan¹^{, 2}, Anisa Fitriyanti¹^{, 2}, Helzi Angelina¹^{, 2}, Gusti Angieta¹^{, 2}, Puan Aqila Azizah¹^{, 2}, Royna Rahma Musie¹^{, 2}, Jefferson Lynford Declan¹^{, 2}, Gladys Indira Putri¹^{, 2}, Siti Fatimah¹^{, 2}, Ayu Berkahingrum¹^{, 2}, Muktiningsih Nurjayadi¹^{, 2}, Fera Kurniadewi¹^{, 2}, Bassam Abomoelak³, Hesham A. El Enshasy⁴^{, 5}^{, 6}

¹Department 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

²Research Center for Detection of Pathogenic Bacteria, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Universitas Negeri Jakarta, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia

³Arnold Palmer Hospital Pediatric Specialty Diagnostic Laboratory, Orlando, FL, 32806, USA

⁴Innovation Center in Agritechnology for Advanced Bioprocessing (ICA), Universiti Teknologi Malaysia (UTM), Pagoh, Johor, Malaysia

⁵School of Chemical and Energy Engineering, Faculty of Engineering, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia

⁶City of Scientific Research and Technology Applications, New Burg Al Arab, Alexandria, Egypt

^*Corresponding author. Email: irmaratna@unj.ac.id

Corresponding Author

Irma Ratna Kartika

Available Online 26 December 2024.

DOI: 10.2991/978-94-6463-624-6_12 How to use a DOI?
Keywords: Enterococcus faecalis; htrA Gene; Pathogen; Detection Method; Real-Time PCR; Bacteria
Abstract: Enterococcus faecalis is known as the opportunistic pathogens that can cause a variety of diseases. These bacteria are capable of producing enterotoxins that lead to gastrointestinal symptoms. This study aims to develop a real-time Polymerase Chain Reaction (PCR) method that is more efficient for identifying Enterococcus faecalis by assessing the results of the confirmation and specificity test of the htrA primer. The concentration of isolated DNA is 107 nanograms per-microliter, and the purities of A260/280 are 2.02. htrA primer successfully amplified Enterococcus faecalis DNA fragments at an annealing temperature of 60 degrees Celsius with an amplicon length of 162 base pairs. These pairs of primers amplified target sequences at a Ct value of 17.62 ± 0.11 and a Tm value of 81.21 degrees Celsius ± 0.06. Specificity tests showed that the htrA primer could distinguish between target and several non-target bacteria. Based on these results, it can be concluded that the Enterococcus faecalis can be efficiently identified by these primers utilizing the real-time PCR. In order to create sensitive and specific detection methods, the next procedure will involve generating htrA primers to identify Enterococcus faecalis in sensitivity testing and artificially contaminated samples.
Open Access: Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

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Volume Title: Proceedings of the 4th Science and Mathematics International Conference (SMIC 2024)
Series: Advances in Physics Research
Publication Date: 26 December 2024
ISBN: 978-94-6463-624-6
ISSN: 2352-541X
DOI: 10.2991/978-94-6463-624-6_12 How to use a DOI?
Open Access: Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

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ris enw bib

TY  - CONF
AU  - Irma Ratna Kartika
AU  - Grace Grace
AU  - Ananda Indah Putri Sihombing
AU  - Agus Setiawan
AU  - Anisa Fitriyanti
AU  - Helzi Angelina
AU  - Gusti Angieta
AU  - Puan Aqila Azizah
AU  - Royna Rahma Musie
AU  - Jefferson Lynford Declan
AU  - Gladys Indira Putri
AU  - Siti Fatimah
AU  - Ayu Berkahingrum
AU  - Muktiningsih Nurjayadi
AU  - Fera Kurniadewi
AU  - Bassam Abomoelak
AU  - Hesham A. El Enshasy
PY  - 2024
DA  - 2024/12/26
TI  - The Potential of htrA Gene to Identify Enterococcus faecalis using real-time Polymerase Chain Reaction
BT  - Proceedings of the 4th Science and Mathematics International Conference (SMIC 2024)
PB  - Atlantis Press
SP  - 129
EP  - 144
SN  - 2352-541X
UR  - https://doi.org/10.2991/978-94-6463-624-6_12
DO  - 10.2991/978-94-6463-624-6_12
ID  - Kartika2024
ER  -

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