The Potential of inlA Gene as a Target Detection for Listeria monocytogenes in Vegetables Sample using Polymerase Chain Reaction Method
Muktiningsih Nurjayadi (a*), Jefferson Lynford Declan (a), Gladys Indira Putri (a), Ismaya Krisdawati (a), Dandy Akbar Juliansyah (a), Maharanianska Azzahra (a), Irvan Maulana (a), Rosita Gio Anggraeni (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, Cipambuan 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

Listeria monocytogenes is a bacteria that causes food poisoning and is found in various types of food. It can infect humans with a mortality rate up to 30 percent. The aim of this study is to determine the potential of internalin A (inlA) gene with Gradient Polymerase Chain Reaction (PCR) as a basis for developing a detection method for Listeria monocytogenes in vegetables using Real-Time PCR. In this study a round white colony was obtained on the Brain Heart Infusion (BHI) agar. In liquid media, the OD600 value is 1.418, which indicates the growth of bacteria. The DNA used as a template had a concentration of 72 ng/micro Liter with A260/280 purity around 1.802. Variation of annealing temperature in Gradient PCR between 53-62 degree Celsius based on melting temperature of the primer. The primer successfully amplifies the inlA gene fragment with an amplicon size 161 base pair. In addition, the optimum annealing temperature for these primers was 60 degree Celsius based on the brightest band and a single band formed in electrophoresis. In the future primers and optimum annealing temperature can be used to develop detection kit for Listeria monocytogenes bacteria in vegetables using Real-Time PCR method.

Keywords: Detection, Foodborne Disease, inlA gene, Listeria monocytogenes, Polymerase Chain Reaction

Topic: Chemistry