Evaluating the Potential of aprA Gene for Accurate Detection of Pseudomonas aeruginosa by real-time Polymerase Chain Reaction
Muktiningsih Nurjayadi1,2, Helzi Angelina1,2, Ananda Indah Putri Sihombing1,2, Puan Aqila Azizah1,2, Anisa Fitriyanti1,2, Royna Rahma Musie1,2, Gusti Angieta1,2, Grace1,2, Agus Setiawan1,2, Jefferson Lynford Declan1,2, Gladys Indira Putri1,2, Dandy Akbar Juliansyah1,2, Siti Fatimah2, Atikah Nur Rahmawati1,2, Irma Ratna Kartika1,2, Fera Kurniadewi1,2, Shyi-Tien Chen3, Bassam Abomoelak4, Hesham A. El Enshasy5,6,7

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.
3Department of Safety, Health and Environmental Engineering, National Kaohsiung University of Science andTechnology, No. 1 University Road, Yanchao District, Kaohsiung City 82445, Taiwan
4 Arnold Palmer Hospital Pediatric Specialty Diagnostic Laboratory, Orlando, FL 32806, USA.
5Innovation Center in Agritechnology for Advanced Bioprocessing (ICA), Universiti Teknologi Malaysia (UTM), Pagoh, Johor, Malaysia.
6School of Chemical and Energy Engineering, Faculty of Engineering, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia.
7City of Scientific Research and Technology Applications, New Burg Al Arab, Alexandria, Egypt.

Abstract

\(Pseudomonas\) \(aeruginosa\) is opportunistic-pathogenic bacteria that cause nosocomial infections. These bacteria that can grow at various temperatures and can be found in diverse environments, enable for \(Pseudomonas\) \(aeruginosa\) to become a contaminant in food during food processing, storage and distribution. This study aims to determine the potential of the \(aprA\) gene primers to detect \(Pseudomonas\) \(aeruginosa\) quick and accurate using the real-time Polymerase Chain Reaction method. The \(aprA\) gene can destroy the microbial recognition molecules thereby reducing the immune of the host response to infection. \(aprA\) can amplify DNA fragments from \(Pseudomonas\) \(aeruginosa\) with an amplicon length of 171 bp at an annealing temperature of 60 degrees Celsius. In the confirmation test, \(aprA\) produced a Ct value of 17.335\(\pm\)0.215 with Tm value of 88.72 degrees Celsius\(\pm\)0.02. \(aprA\) can also differentiate target and non-target bacteria in the specificity test based on diversity in Ct and Tm values. Based on the sensitivity test, \(aprA\) has a detection limit value of 16 picogram per-microliter equivalent to 1.43\(\times\)\(10^4\) CFU with a Ct value of 33.25. Based on these results, \(aprA\) can detect \(Pseudomonas\) \(aeruginosa\) DNA quick and accurate using the rt-PCR method. This research will continue with artificial testing on food.

Keywords: aprA Gene, Detection, Pathogen bacteria, Pseudomonas aeruginosa, rt-PCR

Topic: Chemistry