Optimization fnbA gene target for potential detection Staphylococcus aureus as foodborne pathogen bacteria using Polymerase Chain Reaction Method Muktiningsih Nurjayadi (a*), Maharanianska Azzahra (a), Irvan Maulana (a), Ratna Nur Kusumawati (a), Nabila Alya Pramudiyasih (a), Jefferson Lynford Declan (a), Gladys Indira Putri (a), Ismaya Krisdawati (a), Dandy Akbar Juliansyah (a), Ayu Berkahingrum (a), Irma Ratna Kartika (a), Fera Kurniadewi (a), Dalia Sukmawati (b), Sri Rahayu (b), Vira Saamia (c), Dwi Anna Oktaviani Saputro (c), I Made Wiranatha (c), H. E. Enshay (d)(e)(f)
a) Department 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
*muktiningsih[at]unj.ac.id
b) Department of Biology, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj^ari, 9th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia
c) Center Forensic Laboratory of the Criminal Investigation, Police of the Republic of Indonesia, Ciapmbuan Babakan Madang, Bogor, 1681, Indonesia
d) Institute of Bioproduct Development, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia.
e) School of Chemical and Energy Engineering, Faculty of Engineering, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia.
f) City of Scientific Research and Technology Applications, New Burg Al Arab, Alexandria, Egypt.
Abstract
Staphylococcus aureus can be found in a wide range of areas and emerge as one of pathogenic bacteria that cause foodborne infection. Hence, the optimization for S. aureus detection itself needs to be developed, to produce more accurate results. This study aims to determine the optimal annealing temperature condition and the potential of fnbA primers target gene in the development of S. aureus detection using Polymerase Chain Reaction method. Primer pairs fnbA design was firstly synthesized and followed by the cultural growth process of S. aureus bacteria in MSA agar, resulting in producing yellow colonies. DNA isolate with concentration of 26 ng/ micro Liter and purity 1,8 of S. aureus ATCC 25923 was used as a PCR template. The optimization stage is carried out by varying the temperature at a range of 57-61 degree Celsius resulting in the appearance of annealing bands on agar when exposed to UV light. The results showed that the primer successfully amplified fnbA S. aureus gene with a single band showed the brightnest at temperature 60 degree Celsius and produced an amplicon 187 bp size as targeted. Thus, the optimal conditions that have been obtained can be used for the next stage of detection in food using Real-time PCR.
