Potential of hpmA Gene as a Detection Method of Proteus mirabilis Bacteria using real-time Polymerase Chain Reaction
Muktiningsih Nurjayadi1,2, Royna Rahma Musie1,2, Anisa Fitriyanti1,2, Gusti Angieta1,2, Ananda Indah Putri Sihombing1,2, Puan Aqila Azizah1,2, Helzi Angelina1,2, Grace1,2, Agus Setiawan1,2, Jefferson Lynford Declan1,2, Gladys Indira Putri1,2, Dandy Akbar Juliansyah1,2, Siti Fatimah2, Rosita Gio Anggraeni1,2, Novitasari 3, Irma Ratna Kartika1,2, Fera Kurniadewi1,2, Novitasari 3, 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.
3Research Center for Testing Technology and Standard, National Research and Innovation Agency (BRIN), Jl. Raya Puspitek Serpong, Tangerang Selatan 15314, Indonesia.
4Arnold 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

\(Proteus\) \(mirabilis\) is pathogenic bacteria that can cause gastrointestinal infections, bacteremia, and Urinary Tract Infections (UTI). Therefore, it is necessary to have a fast, sensitive, specific, and accurate detection method to identify \(Proteus\) \(mirabilis\). This study aims to determine the confirmation, specificity, and sensitivity test of the \(hpmA\) gene primer to detect \(Proteus\) \(mirabilis\) quick and precise using the real-time Polymerase Chain Reaction method. Gradient Polymerase Chain Reaction results showed the \(hpmA\) primer has an amplicons length of 195 bp and the optimum annealing temperature at 60 degrees Celsius. The primer pair produced Ct value of 10.40\(\pm\)0.18 and showed one peak in the melting curve with Tm value of 81.84 degrees Celsius \(\pm\)0.02 by real-time PCR. Furthermore, the \(hpmA\) primer was also able to distinguish target and non-target bacteria based on the difference in Ct and Tm value formed. Based on these results, the concentration of bacterial DNA that can be detected by primers reached 3.2 pg/\(\mu\)L, equivalent to the concentration of target bacteria that can be detected by primers is 10.24\(\times\)\(10^{2}\)CFU. In the next step, \(hpmA\) primer will be developed to detect \(Proteus\) \(mirabilis\) in artificial contaminated samples using real-time PCR.

Keywords: Pathogenic bacteria, Detection method, real-time Polymerase Chain Reaction, Proteus mirabilis, hpmA gene

Topic: Chemistry